ERβ促进激素非依赖性前列腺癌细胞凋亡的体内实验研究
摘要
目前已经克隆出两种雌激素受体，ERα与ERβ。在人类前列腺组织中，可同时表达ERα和ERβ这两种受体。ERβ在正常前列腺中表达水平较高，主要分布于前列腺上皮细胞。
研究表明：与正常前列腺组织相比，前列腺癌组织中的ERβ表达减少或缺失，且ERβ的丢失与前列腺癌的发生、发展紧密相关。ERβ表达缺失的研究有助于说明ERβ对于前列腺具有潜在的保护性作用。基因敲除鼠的长期实验发现，ERβ基因敲除鼠（ERβ-/-），可出现前列腺上皮细胞显著增生，而ERα基因敲除鼠（ERα-/-）未见此种情况，提示，ERβ对于前列腺上皮增生具有抑制作用。此外，研究显示：ERβ在前列腺癌细胞中具有抗增殖和促凋亡的作用，ERβ转染前列腺癌细胞能够抑制细胞的增殖，诱导细胞凋亡，并且ERβ转染其他恶性肿瘤如结肠癌、乳腺癌、卵巢癌等细胞同样能够抑制细胞的增殖，诱导细胞凋亡。因此ERβ在肿瘤发生与发展中的作用具有重要重要的研究价值。
我们前期的工作，已通过基因重组技术构建出携带ERβ的pcDNA3.1-ERβ质粒，将其转染至人激素非依赖性前列腺癌PC-3M和PC-3细胞中，结果显示：ERβ能够抑制细胞的增殖、侵袭，诱导细胞发生凋亡，同时发现ERβ能够抑制促生存通路中Akt(蛋白激酶B)的表达。
目前研究表明，Akt在前列腺癌、卵巢癌、乳腺癌、结肠癌等肿瘤中处于活化状态。Akt在细胞中以去磷酸化状态存在，磷酸化后可向细胞质或细胞核转运，与底物蛋白作用，活化其下游靶基因，促进肿瘤细胞生长、血管生成、侵袭和转移，抑制细胞凋亡。
本实验通过构建裸鼠前列腺原位移植癌模型，利用减毒沙门氏菌携带pcDNA3.1-ERβ质粒到小鼠前列腺肿瘤组织内，体内实验观察ERβ对前列腺原位移植癌凋亡的影响。
实验目的：建立裸鼠前列腺原位移植癌模型，应用减毒沙门氏菌携带pcDNA3.1-ERβ质粒，观察ERβ对裸鼠前列腺原位移植癌凋亡的影响。实验方法：构建裸鼠前列腺原位移植癌模型，原位移植术后随机分组：MOCK组给予PBS；PQ组给予减毒沙门氏菌；pcDNA3.1组给予减毒沙门氏菌携带pcDNA3.1空质粒；pcDNA3.1-ERβ组给予减毒沙门氏菌携带pcDNA3.1-ERβ质粒。原位移植手术三天后采用滴鼻给药方式，分别给予相应的10μl PBS及10μl含1×107个菌的各组菌液，第十天后第二次给菌，方式剂量同第一次给菌。观察裸鼠状态及肿瘤的生长情况。待对照组肿瘤长到一定大小时处死全部动物，收集肿瘤样本，AnnexinⅤ和TUNEL法检测肿瘤细胞的凋亡。免疫组化法检测PCNA蛋白的表达水平。Western blot检测ERβ，p-Akt，Bad/Bcl-xl，cleaved caspase9，cleaved caspase3，cleaved PARP的蛋白表达。PCR检测Akt，Bcl-xl，caspase3基因表达。
实验结果：与对照组相比，pcDNA3.1-ERβ组裸鼠恶病质程度明显较低，肿瘤重量减轻，肿瘤生长受到抑制。AnnexinⅤ和TUNEL检测结果发现pcDNA3.1-ERβ组肿瘤细胞出现凋亡细胞。免疫组织化学结果可见pcDNA3.1-ERβ组PCNA蛋白表达下降。Western blot实验显示，与MOCK组，PQ组，pcDNA3.1组相比，pcDNA3.1-ERβ组ERβ，Bad/Bcl-xl，cleaved caspase9，cleaved caspase3及cleavedPARP的蛋白表达上调；而p-Akt的蛋白表达下降。PCR结果显示，与MOCK组，PQ组，pcDNA3.1组相比，pcDNA3.1-ERβ组Akt, Bcl-xl，caspase3基因表达下降。
结论：ERβ能够诱导前列腺原位移植癌细胞的凋亡，表明ERβ与前列腺癌发生密切相关；其诱导凋亡的机制是ERβ抑制了Akt的活性，从而解除了Akt对其下游靶基因Bad的抑制，促凋亡蛋白Bad的活性增加和/或抗凋亡蛋白Bcl-xl减少，激活caspase9和caspase3的级联反应，诱导前列腺癌细胞凋亡。
There are two kinds of ER, estrogen receptor alpha (ERα) and estrogen receptorbeta (ERβ). Both of these two receptors are expressed in human prostate. ERβ showshigh expressed level in normal prostate, and mostly localized to the basal epithelialcompartment of the normal prostate.
But there are growing evidences show that a progression loss of ERβ exprssion inprostate cancer compared with normal prostate, and is associated with normal prostateepithelium developed into prostate cancer. These studies about loss the expression ofERβ during carcinogenesis add to an accumulating body of evidences supporting apotential protective role of ERβ. Findings in ERβ knockout mice indicated that theseanimals develop prostatic hyperplasia in old age, a phenomenon that does not occur inERα knockout mice, this evidence suggestes that ERβ potential protective role inprostate epithelial cells. Furthermore, other studies have shown that ERβ may exert aprotective effect against aberrant cell proliferation and/or carcinogenesis, ERβ isanti-proliferation and proapoptotic in the prostate and also have shown that in coloncancer, breast cancer, ovarian cancer. So, ERβ have an important research value inprostate tumorigenesis and development.
We have constructed the recombination plasmid pcDNA3.1-ERβ which with thehuman estrogen receptor2(ERβ) full length cDNA, and transfected it into humanandrogen-independent prostate cancer cells PC-3M and PC-3. It’s shown that ERβ caninhibit the cells’ proliferation and invasion, induce cells’ apoptosis, and we also foundthat ERβ can inhibit the expression of Akt (protein kinase B) in the pro-survivalpathway.
In current research shows that Akt existed as activated phosphorylation state inprostate cancer, ovarian cancer, breast cancer, colon carcinoma etc., and closely correlated with tumorous genesis and development. Akt existed in cells as adephosphorylation state, and transported to cytoplasm or cell nucleus after activated.It was activated as a phosphorylation state by binding with substrate protein andactivated Akt (p-Akt) can regulate the downstream target gene, promote the growth ofcancer cells, angiogenesis, invasion and metastasis and inhibit the apoptosis.
In this study, by construct the nude mice prostate orthotopin transplantation cancermodel, the attenuated Salmonella carrying pcDNA3.1-ERβ plasmid was given to themice prostate tumor tissue, in vivo experimental observation the effect of ERβinducing prostate orthotopin transplantation cancer apoptosis.
Purpose:
Construct the prostatic orthotopic transplantation cancer models of nude mice,attenuated salmonella carrying pcDNA3.1-ERβ plasmid to observe the effect of ERβon the apoptosis of orthotopic transplantation prostatic cancer in nude mice.
Methods:
The prostatic orthotopic transplantation cancer models of mice were built, themice were randomly divided into MOCK group treated with PBS, PQ group treatedwith attenuated salmonella alone, pcDNA3.1group treated with attenuated salmonellacarrying pcDNA3.1plasmid, pcDNA3.1-ERβ group treated with attenuatedsalmonella carrying pcDNA3.1-ERβ plasmid, three days after operation,10μl PBSand10μl1×107bacteria intranasal drug delivery to give each group respectively, onthe tenth day, the second treatment was given in the first manner. The general states ofmice and the tumors growth were observed. All animals were sacrificed when thetumor in the control group grows to a certain size. Amounts of apoptosis in tumorcells were analyzed with AnnexinⅤ and TUNEL assay. The protein expression levelsof PCNA were analyzed with immunohistochemistry. The protein expression levels ofERβ and p-Akt, Bad/Bcl-xl, cleaved caspase9, cleaved caspase3, cleaved PARP wereanalyzed with western blot. The mRNA expression levels of Akt, Bcl-xl and caspase3were analyzed by PCR.
Results:
Compared with Mock group, PQ group, pcDNA3.1group, the degrees of micecachexia in pcDNA3.1-ERβ group were significantly reduced, the tumor weight weredecreased and the tumor growth were inhibited. The result of AnnexinⅤ and TUNELrevealed that there are apoptosis cells in pcDNA3.1-ERβ group. The result ofimmunohistochemistry revealed that PCNA expression levels in pcDNA3.1-ERβgroup were decreased. Compared with Mock group, PQ group, pcDNA3.1group, theprotein expression levels of ERβ, Bad/Bcl-xl, cleaved caspase9, cleaved caspase3andcleaved PARP in pcDNA3.1-ERβ group were significantly increased (P<0.05), theprotein expression level of p-Akt were decreased (P<0.05). Compared with Mockgroup, PQ group, pcDNA3.1group, the Akt, Bcl-xl and caspase3mRNA expressionlevels were reduced in pcDNA3.1-ERβ group (P<0.05).
Conclusion:
ERβ was able to induce apoptosis of orthotopic implantation prostatic cancercells in mice, indicate that ERβ is closely related to prostate tumorigenesis. Themechanism of induce apoptosis is ERβ inhibits Akt activity, which lead relieves theAkt inhibition its downstream target genes Bad, the increased activity of pro-apoptoticprotein Bad and/or anti-apoptotic protein Bcl-xl reduced, then activation of caspase9and caspase3cascade, inducd the apoptosis of tumor cells.
研究表明：与正常前列腺组织相比，前列腺癌组织中的ERβ表达减少或缺失，且ERβ的丢失与前列腺癌的发生、发展紧密相关。ERβ表达缺失的研究有助于说明ERβ对于前列腺具有潜在的保护性作用。基因敲除鼠的长期实验发现，ERβ基因敲除鼠（ERβ-/-），可出现前列腺上皮细胞显著增生，而ERα基因敲除鼠（ERα-/-）未见此种情况，提示，ERβ对于前列腺上皮增生具有抑制作用。此外，研究显示：ERβ在前列腺癌细胞中具有抗增殖和促凋亡的作用，ERβ转染前列腺癌细胞能够抑制细胞的增殖，诱导细胞凋亡，并且ERβ转染其他恶性肿瘤如结肠癌、乳腺癌、卵巢癌等细胞同样能够抑制细胞的增殖，诱导细胞凋亡。因此ERβ在肿瘤发生与发展中的作用具有重要重要的研究价值。
我们前期的工作，已通过基因重组技术构建出携带ERβ的pcDNA3.1-ERβ质粒，将其转染至人激素非依赖性前列腺癌PC-3M和PC-3细胞中，结果显示：ERβ能够抑制细胞的增殖、侵袭，诱导细胞发生凋亡，同时发现ERβ能够抑制促生存通路中Akt(蛋白激酶B)的表达。
目前研究表明，Akt在前列腺癌、卵巢癌、乳腺癌、结肠癌等肿瘤中处于活化状态。Akt在细胞中以去磷酸化状态存在，磷酸化后可向细胞质或细胞核转运，与底物蛋白作用，活化其下游靶基因，促进肿瘤细胞生长、血管生成、侵袭和转移，抑制细胞凋亡。
本实验通过构建裸鼠前列腺原位移植癌模型，利用减毒沙门氏菌携带pcDNA3.1-ERβ质粒到小鼠前列腺肿瘤组织内，体内实验观察ERβ对前列腺原位移植癌凋亡的影响。
实验目的：建立裸鼠前列腺原位移植癌模型，应用减毒沙门氏菌携带pcDNA3.1-ERβ质粒，观察ERβ对裸鼠前列腺原位移植癌凋亡的影响。实验方法：构建裸鼠前列腺原位移植癌模型，原位移植术后随机分组：MOCK组给予PBS；PQ组给予减毒沙门氏菌；pcDNA3.1组给予减毒沙门氏菌携带pcDNA3.1空质粒；pcDNA3.1-ERβ组给予减毒沙门氏菌携带pcDNA3.1-ERβ质粒。原位移植手术三天后采用滴鼻给药方式，分别给予相应的10μl PBS及10μl含1×107个菌的各组菌液，第十天后第二次给菌，方式剂量同第一次给菌。观察裸鼠状态及肿瘤的生长情况。待对照组肿瘤长到一定大小时处死全部动物，收集肿瘤样本，AnnexinⅤ和TUNEL法检测肿瘤细胞的凋亡。免疫组化法检测PCNA蛋白的表达水平。Western blot检测ERβ，p-Akt，Bad/Bcl-xl，cleaved caspase9，cleaved caspase3，cleaved PARP的蛋白表达。PCR检测Akt，Bcl-xl，caspase3基因表达。
实验结果：与对照组相比，pcDNA3.1-ERβ组裸鼠恶病质程度明显较低，肿瘤重量减轻，肿瘤生长受到抑制。AnnexinⅤ和TUNEL检测结果发现pcDNA3.1-ERβ组肿瘤细胞出现凋亡细胞。免疫组织化学结果可见pcDNA3.1-ERβ组PCNA蛋白表达下降。Western blot实验显示，与MOCK组，PQ组，pcDNA3.1组相比，pcDNA3.1-ERβ组ERβ，Bad/Bcl-xl，cleaved caspase9，cleaved caspase3及cleavedPARP的蛋白表达上调；而p-Akt的蛋白表达下降。PCR结果显示，与MOCK组，PQ组，pcDNA3.1组相比，pcDNA3.1-ERβ组Akt, Bcl-xl，caspase3基因表达下降。
结论：ERβ能够诱导前列腺原位移植癌细胞的凋亡，表明ERβ与前列腺癌发生密切相关；其诱导凋亡的机制是ERβ抑制了Akt的活性，从而解除了Akt对其下游靶基因Bad的抑制，促凋亡蛋白Bad的活性增加和/或抗凋亡蛋白Bcl-xl减少，激活caspase9和caspase3的级联反应，诱导前列腺癌细胞凋亡。
There are two kinds of ER, estrogen receptor alpha (ERα) and estrogen receptorbeta (ERβ). Both of these two receptors are expressed in human prostate. ERβ showshigh expressed level in normal prostate, and mostly localized to the basal epithelialcompartment of the normal prostate.
But there are growing evidences show that a progression loss of ERβ exprssion inprostate cancer compared with normal prostate, and is associated with normal prostateepithelium developed into prostate cancer. These studies about loss the expression ofERβ during carcinogenesis add to an accumulating body of evidences supporting apotential protective role of ERβ. Findings in ERβ knockout mice indicated that theseanimals develop prostatic hyperplasia in old age, a phenomenon that does not occur inERα knockout mice, this evidence suggestes that ERβ potential protective role inprostate epithelial cells. Furthermore, other studies have shown that ERβ may exert aprotective effect against aberrant cell proliferation and/or carcinogenesis, ERβ isanti-proliferation and proapoptotic in the prostate and also have shown that in coloncancer, breast cancer, ovarian cancer. So, ERβ have an important research value inprostate tumorigenesis and development.
We have constructed the recombination plasmid pcDNA3.1-ERβ which with thehuman estrogen receptor2(ERβ) full length cDNA, and transfected it into humanandrogen-independent prostate cancer cells PC-3M and PC-3. It’s shown that ERβ caninhibit the cells’ proliferation and invasion, induce cells’ apoptosis, and we also foundthat ERβ can inhibit the expression of Akt (protein kinase B) in the pro-survivalpathway.
In current research shows that Akt existed as activated phosphorylation state inprostate cancer, ovarian cancer, breast cancer, colon carcinoma etc., and closely correlated with tumorous genesis and development. Akt existed in cells as adephosphorylation state, and transported to cytoplasm or cell nucleus after activated.It was activated as a phosphorylation state by binding with substrate protein andactivated Akt (p-Akt) can regulate the downstream target gene, promote the growth ofcancer cells, angiogenesis, invasion and metastasis and inhibit the apoptosis.
In this study, by construct the nude mice prostate orthotopin transplantation cancermodel, the attenuated Salmonella carrying pcDNA3.1-ERβ plasmid was given to themice prostate tumor tissue, in vivo experimental observation the effect of ERβinducing prostate orthotopin transplantation cancer apoptosis.
Purpose:
Construct the prostatic orthotopic transplantation cancer models of nude mice,attenuated salmonella carrying pcDNA3.1-ERβ plasmid to observe the effect of ERβon the apoptosis of orthotopic transplantation prostatic cancer in nude mice.
Methods:
The prostatic orthotopic transplantation cancer models of mice were built, themice were randomly divided into MOCK group treated with PBS, PQ group treatedwith attenuated salmonella alone, pcDNA3.1group treated with attenuated salmonellacarrying pcDNA3.1plasmid, pcDNA3.1-ERβ group treated with attenuatedsalmonella carrying pcDNA3.1-ERβ plasmid, three days after operation,10μl PBSand10μl1×107bacteria intranasal drug delivery to give each group respectively, onthe tenth day, the second treatment was given in the first manner. The general states ofmice and the tumors growth were observed. All animals were sacrificed when thetumor in the control group grows to a certain size. Amounts of apoptosis in tumorcells were analyzed with AnnexinⅤ and TUNEL assay. The protein expression levelsof PCNA were analyzed with immunohistochemistry. The protein expression levels ofERβ and p-Akt, Bad/Bcl-xl, cleaved caspase9, cleaved caspase3, cleaved PARP wereanalyzed with western blot. The mRNA expression levels of Akt, Bcl-xl and caspase3were analyzed by PCR.
Results:
Compared with Mock group, PQ group, pcDNA3.1group, the degrees of micecachexia in pcDNA3.1-ERβ group were significantly reduced, the tumor weight weredecreased and the tumor growth were inhibited. The result of AnnexinⅤ and TUNELrevealed that there are apoptosis cells in pcDNA3.1-ERβ group. The result ofimmunohistochemistry revealed that PCNA expression levels in pcDNA3.1-ERβgroup were decreased. Compared with Mock group, PQ group, pcDNA3.1group, theprotein expression levels of ERβ, Bad/Bcl-xl, cleaved caspase9, cleaved caspase3andcleaved PARP in pcDNA3.1-ERβ group were significantly increased (P<0.05), theprotein expression level of p-Akt were decreased (P<0.05). Compared with Mockgroup, PQ group, pcDNA3.1group, the Akt, Bcl-xl and caspase3mRNA expressionlevels were reduced in pcDNA3.1-ERβ group (P<0.05).
Conclusion:
ERβ was able to induce apoptosis of orthotopic implantation prostatic cancercells in mice, indicate that ERβ is closely related to prostate tumorigenesis. Themechanism of induce apoptosis is ERβ inhibits Akt activity, which lead relieves theAkt inhibition its downstream target genes Bad, the increased activity of pro-apoptoticprotein Bad and/or anti-apoptotic protein Bcl-xl reduced, then activation of caspase9and caspase3cascade, inducd the apoptosis of tumor cells.
引文
[1] Mosselman S, Polman J, Dijkema R.ER beta: identification and characterizationof a novel human estrogen receptor. FEBS Lett.1996;392(1):49-53.
[2] Peach K, Webb P, Kuiper G, et al.Different ligand activation of estrogen receptorsERα and ERβ at AP-1site.Science1997;277:1508-1510.
[3] Onate SA, Boonyaratanakornkit V, Spencer TE, et al.The steroid receptorcoactivator-1contains multiple receptor interacting and activation domains thatcooperatively enhance the activation function1(AF-1) and AF-2domains ofsterodi receptors. Journal of Biological Chemistry1998;22:5011-5020
[4] Barkhem T, Carlsson B, Nilsson Y, et al.Differential response of estrogen receptoralpha and estrogen beta to partial estrogen agonists/antagonists.MolecularPharmacology1998;54:105-112.
[5] Gustafsson, JA.What pharmacologists can learn from recent advances in estrogensignalling.Trends Pharmacol.Sci2003;24:479-485.
[6] Cook LS, Goldoft M, Schwartz SM, et al.Incidence of adenocarcinoma of theprostate in Asian immigrants to the United States and their descendants. J Urol1999;161:152-155.
[7] K. Griffiths, International Prostate Health Council Study Group.Estrogens andprostatic cancer.Prostate2000;45:87-100.
[8] Liu MM, Albanese C, Anderson CM, et al.Opposing action of estrogen receptorsalpha and beta on cyclin D1gene expression.J Biol Chem2002;277:24353-24360.
[9] Shapiro E, Huang H, Masch RJ, et al.Immunolocalization of estrogen receptoralpha and beta in human fetal prostate.J. Urol.2005;174:2051-2053.
[10]Bonkhoff H, Fixemer T, Hunsicker I, et al.Estrogen receptor expression inprostate cancer and premalignant prostatic lesions. Am J Pathol1999;155:641-647.
[11]Leav I, Lau KM, Adams JY, et al.Comparative studies of the estrogen receptoes-aand-b and the androgen receptor in normal human prostate glands, dysplasia, andin primary and metastatic carcinoma. Am J Pathol2001;159:79-92.
[12]Latil KM, Bieche I, Vidaud D, et al.Evaluation of Androgen, Estrogen(ERα andERβ), and Progesterone Receptor Expression in Human Prostate Cancer byReal-Time Quantitative reverse Transcription-Polymerase Chain Reactionassays.Cancer Reaserch2001;61:1919-1926.
[13]Weihua Z, Lathe R, Warner M, et al.An endocrine pathway in the prostate, ER-β,AR,5-α-androstane-3β,17β-diol, and CYP7B1, regulates prsotate growth.ProcNatl Acad Sci USA2002;99:13589-13594.
[14]Pasquali D, Staibano S, Prezioso D, et al.Estrogen receptorβexpression in humanprostate tissue. Molecular and Cellular Endocrinology2001a;178:47-50.
[15]Horvath LG, Henshall SM, Lee CS, et al.Frequent loss of estrogen receptor-bexpression in prostate cancer. Cancer Res2001;61:5331-5335.
[16]Fujimura T, Takahashi S, Urano T, et al.Different expression of estrogen receptorbeta(ERbeta) and its C-terminal truncated splice variant ER betacx as prognosticpredictors in human prostatic cancer.Biochem Biophs Res Commun2001;289:692-699.
[17]Weihua Z, Warner M, Gustafsson JA.Estrogen receptor beta in the prostate.MolCell Endocrinol2002;193:1–5.
[18]Torlakovic E, Lilleby W, Torlakovic G, et al.Prostate carcinoma expression ofestrogen receptor-beta as detected by PPG5/10antibody has positive associationwith primary Gleason grade and Gleason score.Hum Pathol2002;33:646-651.
[19]Fixemer T, Remberger K, Bonkhoff H.Differential expression of the estrogenreceptor beta(ER-b) in human prostate tissue, premalignant changes, and inprimary, metastatic, and recurrent prostatic adenocarcinoma. Prostate2003;54:79-87.
[20]Ji Q, Liu PI, Elshimali Y, et al.Frequent loss of estrogen and progesteronereceptors in human prostatic tumors determined by quantitative real-timePCR.Mol Cell Endocrinol2005;229:103-110.
[21]Yang Guo-sheng, Wang Ying, Wang Ping et al.Expression of oestrogen receptor-αand oestrogen receptor-β in prostate cancer.Chinese Medical Journal2007;120:1611-1615.
[22]Margaret Warner, Jan-Ake Gustafsson.The role of estrogen receptor β(ERβ)inmalignant diseases-A new potential target for antiproliferative drugs in preventionand treatment of cancer.Biochemical and Biophsical Research Communications2010;396:63-65.
[23]Asgai and Morakabat.Estrogen receptor beta expression in prostate adenocarcinoma.Diagnostic Pathology2011;6:61.
[24]Nojima D, Li LC, Dharia A, et al.CpG hypermethylation of the promoter regioninactivates the estrogen receptor-beta gene in patients with prostate carcinoma.Cancer2001;92:2076-2083.
[25]Sasaki M, Tanaka Y, Perinchery G, et al.Methylation and inactivation of estrogen,progesterone, and androgen receptors in prostate cancer. J Natl Cancer Inst2002;94:384-390.
[26]Zhu X, Leav I, Leung YK, et al.Dynamic regulaiton of estrogen receptor-betaexpression by DNA methylation during prostate cancer development andmetasis.Am J Pathol2004;164:2003-2012.
[27]Lau KM, LaSpina M, Long J, et al.Expression of estrogen receptor(ER)-alphaand ER-beta in mormal and malignant prostatic epithelial cells: regulation bymethylation and involvement in growth regulaiton.Cance Res2000;60:3175-3182.
[28]Jarred, RA, Cancilla, B, Prins, GS, et al.Evidence that estrogens directly alterandrogen-regulated prostate development.Endocrinology2000;141:3471-3477.
[29]Weihua Z, Makela S, Andersson LC, et al.A role for estrogen receptor β in theregulation of growth of the ventral prostate.Proc Natl Acad Sci USA2001;98:6330-6335.
[30]Imamov O, Morani A, Shim GJ, et al.Estrogen receptor β regulates epithelialcellular differentiation in the mouse ventral prostate. Proc Natl Acad Sci USA2004;101:9375-9380.
[31]Jennifer Cheng, Eun Jig Lee, Laird D. Madison, et al.Expression of estrogenreceptor β in prostate carcinoma cells inhibits invasion and proliferatioin andtriggers apoptosis.FFBS Letters2004;566:169-172.
[32]Neubauer BL, McNulty AM, Chedid M, et al.The selective estrogen receptormodulator trioxifene (LY133314) inhibits metastasis and extends survival in thePAIII rat prostatic carcinoma model.Cancer Res2003;63:6056-6062.
[33]Montano, M.M., H. Deng, M. Liu, et al.Transcriptional regulation by the estrogenreceptor of antioxidative stress enzymes and its functional implications.Oncogene2004;23:2442-2453.
[34]Guerini V, Sau D, Scaccianose E, et al.The androgen derivative5alpha-androstane-3beta,17beta-diol inhibits prostate cancer cell migration throughactivation of the estrogen receptor beta subtype.Cancer Res2005;65:5445-5453.
[35]Paul Mak, Yuet-Leung, Wan-Yee Tang, et al.Apigenin suppresses cancer cellgrowth through ERβ.Neoplasia2006;8:896-904.
[36]Ellem SJ, Risbridger GP.Treating prostate cancer: A rationale for targeting localoestrogens.Nature Rev Cancer2007;7:621-627.
[37]McPherson SJ, Ellem SJ, Simpson ER, et al.Essential role for estrogen receptorbeta in stromal-epithlial regulation of prostatic hyperplasia.Endocrinology2007;148:566-574.
[38]Pravettoni A, Mornati O, Martini PG, et al.Estrogen receptor beta (ERbeta) andinhibition of prostate cancer cell proliferation: Studies on the possible mechanismof action in DU145cells.Mol Cell Endocrinol2007;263:46-54.
[39]Stettner M, Kaulfuss S, Burfeind P, et al.The relevance of estrogen receptor-betaexpression to the antiproliferative effects observed with hitone deacetylaseinhibitors and phytoestrogens in prostate cancer treatment.Mol Cancer Ther2007;6:2626-2633.
[40]Bonhoff H, Berges R.The evolving role of oestrogens and their receptors in thedevelopment and progression of prostate cancer.Eur Urol2008;55:533-542.
[41]Kentaro Matsumura, Tomokat Tanaka, Hidenori Kawashima, et al.Involvement ofthe estrogen receptor β in Genistein-induced expression of p21waf1/cip1in PC-3prostate cancer cells.ANTICANCER RESEARCH2008;28:709-714.
[42]Walton TJ, Li G, Seth R, et al.DNA Demethylation and histone deacetylationinhibition co-Operate to re-express estrogen receptor beta and induec apoptosis inprostate cancer cell-lines.The Prostate2008;68:210-222.
[43]D.M.A. Attia and A.G..H. Ederveen.Opposing Roles of ERαand ERβ in theGenesis and Progression of Adenocarcinoma in the Rat Ventral Prostate.TheProstate2011.
[44]Takahashi Y, Lavigne JA, Hursting SD, et al.Molecular signatures of soy-derivedphytochemicals in androgen-responsive prostate cancer cells: a comparison studyusing DNA microarray.Mol Carcinog2006;45:943-956.
[45]Ronald L. Shazer, Anjali Jain, Anna V. Galkin, et al.Raloxifene, an oestrogen-receptor-β-targeted therpy, inhibits androgen-independent prostate cancergrowth:results from preclinical studies and a pilot phaseⅡ clinical trial.BJUInternational2006;97:691-697.
[46]Kim IY, Kim BC, Seong DH, et al.Raloxifene a mixed estrogen agonist/antagonist, induces apoptosis in androgen-independent human prostate cancer celllines. Cancer Res2002a;62:5365-5369.
[47]Rossi V, Bellastella G, Rosa CD, et al.Raloxifene induces cell death and inhibitsproliferation through multiple signaling pathways in prostate cancer cells.J. Cell.Physio.2011;226:1334-1339.
[48]Shen JC, Klein RD, Wei Q, et al.Low-dose genistein induces cyclin-dependentkinase inhibitors and G1cell-cycle arrest in human prostate cancer cells.MolCarcinog2000;29:92-102.
[49]Mentor-Marcel R, Lamaritiniere CA, Eltoum IE, et al.Genistein in the dietreduces the incidence of poorly differentiated prostatic adenocarcinoma intransgenic mice(TRAMP).Cancer Res.2001;61(18):6777-6782.
[50]Mak P. Leung YK, Tang WY, Harwood C, et al.Apigenin supresses cancer cellgrowth through ERbeta.Neoplasia2006;8:896-904.
[51]Skliris GP, Munot K, Bell SM, et al.Reduced expression of estrogen receptor β ininvasive breast cancer and its reexpression using DNA methyltransferaseinhibitors in a cell line mode.J Pathol2003;201:213
[52]Rody A, Holtrich U, Solbach C, et al.Methylation of estrogen receptor β promotercorrelates with loss of ERβ expression in mammary carcinoma and is an earlyindication marker in premalignant lesions.Endoc Relat Cancer2005;12:903.
[53]Zhao C, Lam EW, Sunters A, et al.Expression of estrogen receptor beta isforms innormal breast epithelial cells and breast cancer: regulaiton by methylation.Oncogene2003;22:7600-7606.
[54]Omoto Y, Inoue S, Ogawa S, et al.Clinical value of the wild-type estrogenreceptor β expression in breast cancer.Cancer Letters2001;163:207-212.
[55]Lindberg K, Strom A, Lock J G, et al.Expression of estrogen receptor β increasesintegrin α1and integrin β1levels and enhances adhesion of breast cancer cells. JCell Physiol2010;222:156-167.
[56]Lindberg et al. Estrogen receptor β represses Akt signaling in breast cancer cellsvia downregulation of HER2/HER3and upregulation of PTEN: implications fortamoxifen sensitivity. Breast Cancer Research2011;13:R43
[57]Lindgen P, Cajiander S, Backstrom T, et al.Estrogen and progesterone andprogeterone recptors in epithelial ovarian tumors.Mol Cell Endocrinol2004;22:97-104.
[58]Suzuki F, Akahira J, Miura I, et al.Loss of estrogen receptor β isoform expressionand its correlation with aberrant DNA methylation of the5’-untranslated regionin human epithelial ovarian carcinoma.Cancer Sci2008;99:2365-2372.
[59]Treeck O, Pfeiler G, Mitter D, et al.Estrogen receptor β exerts anti-tumoral effectson SK-OV-3ovarian cancer cells.Endocrinol2007;193:421.
[60]Lazennec G.Estrogen recptor beta, a possible tumor suppressor involved inovarian carcinogenesis.Cancer Lett2006;231:151-157.
[61]Campbell-Thompson M, Lynch J&Bhardwaj B.Expression of estrogenreceptor(ER) subtypes ER beta isforms in colon cancer. Cancer Research2001;61:632-640.
[62]Jassam N, Bell SM, Speirs V, et al.Loss of expression of oestrogen receptor betain colon cancer and its association with Dukes’ staging.Oncol Rep2005;14:17-21.
[63]Omoto Y, Eguchi H, Yamamoto-Yamaguchi Y,et al.Estrogen receptor(ER)β1andERβcx/β2inhibits ERα function differently in breast cancer cell lineMCF7.Oncogene2003;22:5011-5020.
[64]Paruthiyil S, Parmar H, Kerekatte V, et al.Estrogen receptor beta inhibits humanbreast cancer cell proliferation and tumor formation by causing a G2cell cyclearrest.Cancer Research2004;64:423-428.
[65]McPherson SJ, Wang H, Jones ME, et al.Elevated androgens and prolactin inaromatase-deficient mice cause enlargement, but not malignancy, of the prostategland.Endocrinology2001;142:2458-2467.
[66]Kypianou N, English HF, Davidson NE, et al.Programmed cell death duringregression of the MCF-7human breast cancer following estrogen ablation.CancerResearch1991;51:162-166.
[67]Perillo B, Sasso A, Abbondanza C, et al.17β-Estradiol inhibits apoptosis inMCF-7cells, inducing bcl-2expression via two estrogen-responsive elementspresent in the coding sequence.Molecular and Cellular Biology2000;20:2890-2901.
[68]Choi KC, Kang SK, Tai CJ, et al.Estrodiol up-regulates antiapoptotic Bcl-2messenger ribonucleic acid and protein in tumorigenic ovarian surface epitheliumcells.Endocrinology2001;142:2351-2360.
[69]Nilsen J, Mor G&Naftolin F.Estrogen-regulated developmental neuronalapoptosis is determined by estrogen receptor subtype and the Fas/Fas ligandsystem.Journal of Neurobiology2000;43:64-78.
[70]Bardin A, Hoffmann P, Boulle N, et al.Involvement of estrogen receptor β inovarian carcinogenesis. Cancer Research2004;64:5861-9.
[71]Qiu Y, Waters CE, Lewis AE, et al.Qestrogen-induced apoptosis in colonocytesexpression oestrogen receptor beta.J Endocrinol2002;174:369-77.
[72]Hsu HH, Cheng SF, Wu CC, et al.Apoptotic effects of over-expressed estrogenreceptor-beta on LoVo colon cancer cell is mediated by p53signaling in aligand-dependent manner.Chin J Physiol2006;49:110-116.
[73]Lazennec G, Bresson D, Lucas A, et al.ERβ inhibits proliferation and invasion ofbreast cancer cells.Endocrinology2001;142:4120-30.
[74]Treeck O, Pfeiler G, Mitter D, et al.Estrogen receptor β1exerts antitumoral effectson SK-OV-3ovarian cancer cells.J Endocrinol2007;193:421-433.
[75]苗丽君,王静. Akt与肿瘤的研究进展[J].国外医学.生理、病理科学与临床分册.2004;24:406-409.
[76]Milas I, Komaki R, Hachiya T, et al.Epidermal growth factor receptor,Cyelooxyenase-2, and BAX expression in the Primary nonsmall cell lung cancerand brain metastases.Clin Cancer Res2003;9:1070-1076.
[77]Liao Y, Grobholz R, Abel U, et al.Increase of AKT/PKB expression corrlates withgleason pattern in human prostate cancer.Int J Cancer2003;107:676-680.
[78]Ayala G, Thompson T, Yang G, et al.High level of phosphory-lated form of Akt-1in prostate cancer and non-neoplastic prostate tissues are strong predictors ofbiochemical recurrence. Clin Can-cer Res2004;10(4):6572-6578.
[79]Kreisberg J, Malik S, Prihoda T, et al. Phosphorylation of Akt (Ser473) is anexcellent preditor of poor clinical outcome in prostate cancer. Cancer Res2004;64(4):5232-5236.
[80]孙丽梅.乳腺癌信号转到途径中Akt激活及意义。中华肿瘤防治杂志，2006;13:1232-1234.
[81]Yamamoto S, Tomita Y, Hoshida Y, et al.Prognostic significance of activated Aktexpression in pancreatic ductal adenocarcinoma.Clin Cancer Res.2004;10:2846-2850.
[82]Balsara BR, Pei J, Mitsuuchi Y, et al.Frequent activation of AKT in non-small celllung carcinomas and preneoplastic bronchial lesions. Cancinogenesis2004;25:2053-2059.
[83]Robertson CN, Roberson KM, Padilla GM, et al.Induction of apoptosis bydithylstilbestrol in hormone-insensitive prostate cancer cells.J Natl Cancer Inst1996;88:908-917.
[84]Signoretti S, Loda M.Estrogen receptor beta in prostate cancer: Brake pedal oraccelerator? Am J Pathol2001;159:13-16.
[85]李峰等，hERβ全长基因表达载体EGFP-C1-hERβ的构建及在PC-3M细胞中的表达。《吉林大学学报（医学版）》，2006;32:189-193.
[86]李峰等,雌激素受体2对前列腺癌细胞系PC-3M增殖的影响.中国病理生理学杂志，2009;25:656-660.
[87]Iwao K, Miyoshi Y, Egawa C, et al.Quantitative analysisi of estrogen receptor-βmRNA and its variants in human breast cancers.International Jounal of Cancer2000;88:733-736.
[88]Theys J, Barbe S, Landuyt W, et al. Tumor-specific gene delivery usinggenetically engineened bacteria. Cmr Gene Ther2003;3:207-221.
[89]Ming Zhao, Meng Yang, Huaiyu Ma, et al Targeted Therapy with a SalmonellaTyphimurium Leucine-Arginine Auxotroph Cures Orthotopic Human BreastTumors in Nude Mice. Cancer Res2006;66:7647-7652.
[90]Angelakopoulos H, Hohomann EL.Pilot study of phoP/phoQ-deleted Salmonellaenterica serovar Typhimurium expressiong Helicobacter pylori urease in adultvolunteers.Infect Immun2000;68:213-41.
[91]Ling Zhang, Lifang Gao, Lijuan Zhao, et al.Intratumoral delivery and suppressionof prostate tumor growth by attenuated salmonella enterica serovar typhimuriumcarrying plasmid-based small interfering RNAs.[J].Cancer Res,2007;67(12):5859-5864.
[92]Hartman J, Lindeberg K, Morani A, et al.Estrogen receptor beta inhibitsangiogenesis and growth of T47D breast cancer xeografts.Cancer Res2006;66:11207-13.
[93]Qin XG, Hua Z, Shuang W, et al.Effects of matrine on Hep G2cell proliferationand expression of tumor relevant proteins in vitro.Pharm Biol2010;48:275-281.
[94]赖建生,夏强,张孝斌,等.增殖细胞核抗原和p27在前列腺增生和前列腺癌组织中的表达及意义.中华肿瘤杂志2004;26:476-478.
[95]Graff J, Konicek B, McNulty A, et al.Increased AKT activity contributes toprostate cancer progression by dramatically accelerating prostate tumor growthand diminishing p27Kip1expression. J Biol Chem2000;275(2):24500-24505.
[96]Sastry KS, Smith AJ, Karpova Y, et al.Diverse anti-apoptotic signaling pathwaysactivated by VIP, EGF and PI3K in prostate cancer cells converge on BAD.J BiolChem2006;281:20891-901.
[97]Giulia Pinton, Warren Thomas, Palolo Bellini, et al.Estrogen receptor exsertstumor repressive functions in human malignant pleural mesothelioma EGFRinactivation and affects response to Gefitinib.PloS ONE2010;11:e14110.
[98]Chang F, Lee JT, Navolanic PM, et al.Involement of PI3-K/Akt pathway in cellcycle progression, apoptosis, and neoplastic transformation: a target for cancerchemotherapy. Leukem ia2003;17(3):590-603.
[99]Chin YR, Toker A.Function of Akt/PKB signaling to cell motility, invasion andthe tumor stromer in cancer[J]. Cell Signaling2009;21(2):470-476.
[100]Hu W, Kavanagh JJ.Anticancer therpy targeting the apoptic pathway.LancetOncle2003;4:721-729.
[101]Cheg EH, Wei MC, Weiler S, et al.BCL-2, BCL-X(L) sequesters BH3domin-only molecules preventing BAX-and-BAK-mediated mitochondrialapoptosis. Mol Cell2001;8:70511.
[102]Wei MC, Zong WX, Cheng EH, et al.Proapoptotic BAX and BAK: a requisitegate-way to mitochondrial dysfunction and death. Science2001;292:727-30.
[103]Choi, SI, Joo SS, Yoo YM.Melationin prevents nitric oxide-induced apoptosisby increasing the interaction between14-3-3[beta] and p-Bad in SK-N-MC cells.Journal of Pineal Research2008;44(1):95-100.
[104]Gonzalez-Garcia M, Perez-Ballestro R, Ding L, et al.Bcl-x is the major Mrnafrom expressed during murine development and its product localizes to themitochondrua. Development1994;120:3033.
[105]Simoes Wust AP, Olie RA, Gautschi O, et al.Bcl-xl antisense treatment induesapoptosis in breast carcinoma cells. Int J Cancer,2000;87(4):582-590.
[106]Kirkin V, Joos S, Zorning M.The role of Bcl-2family menbers in tumorigenesis.Biochim.Biphs. Acta2004;1644:229-249.
[107]Stepien A, Izdebska M, Grzanka A.The types of cell death.Postepy Hig MedDosw2007;61:420-428.
[108]Wang ZB, Liu YQ, Cui YF.Pathways to caspase activation.Cell Biol. Int.2005;29:489-496.
[109]Zhou HL, Li XM, Meinkoth J, et al.Akt Regulates Cell Survival and Apoptosisat a Postmitochondrial Level. J Cell Biol2000;151(3):483-494.
[110]Hu W, Kavanagh JJ.Anticancer therpy targeting the apoptic pathway.LancetOncle2003;4:721-729.
[111]A Ivana Scovassa, Marc Diederich.Modulation of Poly (ADP-ribosylation) inapoptotic cells.Biochemical pharmacology2004;5:1041-1247.
[112]Coopersmith CM, Chang KC, Swanson PE, et al.Overexpression of bcl-2in theintestinal epthithelium imoroves survival in septic mice. Crit Care Med2002;30:195-201.
[2] Peach K, Webb P, Kuiper G, et al.Different ligand activation of estrogen receptorsERα and ERβ at AP-1site.Science1997;277:1508-1510.
[3] Onate SA, Boonyaratanakornkit V, Spencer TE, et al.The steroid receptorcoactivator-1contains multiple receptor interacting and activation domains thatcooperatively enhance the activation function1(AF-1) and AF-2domains ofsterodi receptors. Journal of Biological Chemistry1998;22:5011-5020
[4] Barkhem T, Carlsson B, Nilsson Y, et al.Differential response of estrogen receptoralpha and estrogen beta to partial estrogen agonists/antagonists.MolecularPharmacology1998;54:105-112.
[5] Gustafsson, JA.What pharmacologists can learn from recent advances in estrogensignalling.Trends Pharmacol.Sci2003;24:479-485.
[6] Cook LS, Goldoft M, Schwartz SM, et al.Incidence of adenocarcinoma of theprostate in Asian immigrants to the United States and their descendants. J Urol1999;161:152-155.
[7] K. Griffiths, International Prostate Health Council Study Group.Estrogens andprostatic cancer.Prostate2000;45:87-100.
[8] Liu MM, Albanese C, Anderson CM, et al.Opposing action of estrogen receptorsalpha and beta on cyclin D1gene expression.J Biol Chem2002;277:24353-24360.
[9] Shapiro E, Huang H, Masch RJ, et al.Immunolocalization of estrogen receptoralpha and beta in human fetal prostate.J. Urol.2005;174:2051-2053.
[10]Bonkhoff H, Fixemer T, Hunsicker I, et al.Estrogen receptor expression inprostate cancer and premalignant prostatic lesions. Am J Pathol1999;155:641-647.
[11]Leav I, Lau KM, Adams JY, et al.Comparative studies of the estrogen receptoes-aand-b and the androgen receptor in normal human prostate glands, dysplasia, andin primary and metastatic carcinoma. Am J Pathol2001;159:79-92.
[12]Latil KM, Bieche I, Vidaud D, et al.Evaluation of Androgen, Estrogen(ERα andERβ), and Progesterone Receptor Expression in Human Prostate Cancer byReal-Time Quantitative reverse Transcription-Polymerase Chain Reactionassays.Cancer Reaserch2001;61:1919-1926.
[13]Weihua Z, Lathe R, Warner M, et al.An endocrine pathway in the prostate, ER-β,AR,5-α-androstane-3β,17β-diol, and CYP7B1, regulates prsotate growth.ProcNatl Acad Sci USA2002;99:13589-13594.
[14]Pasquali D, Staibano S, Prezioso D, et al.Estrogen receptorβexpression in humanprostate tissue. Molecular and Cellular Endocrinology2001a;178:47-50.
[15]Horvath LG, Henshall SM, Lee CS, et al.Frequent loss of estrogen receptor-bexpression in prostate cancer. Cancer Res2001;61:5331-5335.
[16]Fujimura T, Takahashi S, Urano T, et al.Different expression of estrogen receptorbeta(ERbeta) and its C-terminal truncated splice variant ER betacx as prognosticpredictors in human prostatic cancer.Biochem Biophs Res Commun2001;289:692-699.
[17]Weihua Z, Warner M, Gustafsson JA.Estrogen receptor beta in the prostate.MolCell Endocrinol2002;193:1–5.
[18]Torlakovic E, Lilleby W, Torlakovic G, et al.Prostate carcinoma expression ofestrogen receptor-beta as detected by PPG5/10antibody has positive associationwith primary Gleason grade and Gleason score.Hum Pathol2002;33:646-651.
[19]Fixemer T, Remberger K, Bonkhoff H.Differential expression of the estrogenreceptor beta(ER-b) in human prostate tissue, premalignant changes, and inprimary, metastatic, and recurrent prostatic adenocarcinoma. Prostate2003;54:79-87.
[20]Ji Q, Liu PI, Elshimali Y, et al.Frequent loss of estrogen and progesteronereceptors in human prostatic tumors determined by quantitative real-timePCR.Mol Cell Endocrinol2005;229:103-110.
[21]Yang Guo-sheng, Wang Ying, Wang Ping et al.Expression of oestrogen receptor-αand oestrogen receptor-β in prostate cancer.Chinese Medical Journal2007;120:1611-1615.
[22]Margaret Warner, Jan-Ake Gustafsson.The role of estrogen receptor β(ERβ)inmalignant diseases-A new potential target for antiproliferative drugs in preventionand treatment of cancer.Biochemical and Biophsical Research Communications2010;396:63-65.
[23]Asgai and Morakabat.Estrogen receptor beta expression in prostate adenocarcinoma.Diagnostic Pathology2011;6:61.
[24]Nojima D, Li LC, Dharia A, et al.CpG hypermethylation of the promoter regioninactivates the estrogen receptor-beta gene in patients with prostate carcinoma.Cancer2001;92:2076-2083.
[25]Sasaki M, Tanaka Y, Perinchery G, et al.Methylation and inactivation of estrogen,progesterone, and androgen receptors in prostate cancer. J Natl Cancer Inst2002;94:384-390.
[26]Zhu X, Leav I, Leung YK, et al.Dynamic regulaiton of estrogen receptor-betaexpression by DNA methylation during prostate cancer development andmetasis.Am J Pathol2004;164:2003-2012.
[27]Lau KM, LaSpina M, Long J, et al.Expression of estrogen receptor(ER)-alphaand ER-beta in mormal and malignant prostatic epithelial cells: regulation bymethylation and involvement in growth regulaiton.Cance Res2000;60:3175-3182.
[28]Jarred, RA, Cancilla, B, Prins, GS, et al.Evidence that estrogens directly alterandrogen-regulated prostate development.Endocrinology2000;141:3471-3477.
[29]Weihua Z, Makela S, Andersson LC, et al.A role for estrogen receptor β in theregulation of growth of the ventral prostate.Proc Natl Acad Sci USA2001;98:6330-6335.
[30]Imamov O, Morani A, Shim GJ, et al.Estrogen receptor β regulates epithelialcellular differentiation in the mouse ventral prostate. Proc Natl Acad Sci USA2004;101:9375-9380.
[31]Jennifer Cheng, Eun Jig Lee, Laird D. Madison, et al.Expression of estrogenreceptor β in prostate carcinoma cells inhibits invasion and proliferatioin andtriggers apoptosis.FFBS Letters2004;566:169-172.
[32]Neubauer BL, McNulty AM, Chedid M, et al.The selective estrogen receptormodulator trioxifene (LY133314) inhibits metastasis and extends survival in thePAIII rat prostatic carcinoma model.Cancer Res2003;63:6056-6062.
[33]Montano, M.M., H. Deng, M. Liu, et al.Transcriptional regulation by the estrogenreceptor of antioxidative stress enzymes and its functional implications.Oncogene2004;23:2442-2453.
[34]Guerini V, Sau D, Scaccianose E, et al.The androgen derivative5alpha-androstane-3beta,17beta-diol inhibits prostate cancer cell migration throughactivation of the estrogen receptor beta subtype.Cancer Res2005;65:5445-5453.
[35]Paul Mak, Yuet-Leung, Wan-Yee Tang, et al.Apigenin suppresses cancer cellgrowth through ERβ.Neoplasia2006;8:896-904.
[36]Ellem SJ, Risbridger GP.Treating prostate cancer: A rationale for targeting localoestrogens.Nature Rev Cancer2007;7:621-627.
[37]McPherson SJ, Ellem SJ, Simpson ER, et al.Essential role for estrogen receptorbeta in stromal-epithlial regulation of prostatic hyperplasia.Endocrinology2007;148:566-574.
[38]Pravettoni A, Mornati O, Martini PG, et al.Estrogen receptor beta (ERbeta) andinhibition of prostate cancer cell proliferation: Studies on the possible mechanismof action in DU145cells.Mol Cell Endocrinol2007;263:46-54.
[39]Stettner M, Kaulfuss S, Burfeind P, et al.The relevance of estrogen receptor-betaexpression to the antiproliferative effects observed with hitone deacetylaseinhibitors and phytoestrogens in prostate cancer treatment.Mol Cancer Ther2007;6:2626-2633.
[40]Bonhoff H, Berges R.The evolving role of oestrogens and their receptors in thedevelopment and progression of prostate cancer.Eur Urol2008;55:533-542.
[41]Kentaro Matsumura, Tomokat Tanaka, Hidenori Kawashima, et al.Involvement ofthe estrogen receptor β in Genistein-induced expression of p21waf1/cip1in PC-3prostate cancer cells.ANTICANCER RESEARCH2008;28:709-714.
[42]Walton TJ, Li G, Seth R, et al.DNA Demethylation and histone deacetylationinhibition co-Operate to re-express estrogen receptor beta and induec apoptosis inprostate cancer cell-lines.The Prostate2008;68:210-222.
[43]D.M.A. Attia and A.G..H. Ederveen.Opposing Roles of ERαand ERβ in theGenesis and Progression of Adenocarcinoma in the Rat Ventral Prostate.TheProstate2011.
[44]Takahashi Y, Lavigne JA, Hursting SD, et al.Molecular signatures of soy-derivedphytochemicals in androgen-responsive prostate cancer cells: a comparison studyusing DNA microarray.Mol Carcinog2006;45:943-956.
[45]Ronald L. Shazer, Anjali Jain, Anna V. Galkin, et al.Raloxifene, an oestrogen-receptor-β-targeted therpy, inhibits androgen-independent prostate cancergrowth:results from preclinical studies and a pilot phaseⅡ clinical trial.BJUInternational2006;97:691-697.
[46]Kim IY, Kim BC, Seong DH, et al.Raloxifene a mixed estrogen agonist/antagonist, induces apoptosis in androgen-independent human prostate cancer celllines. Cancer Res2002a;62:5365-5369.
[47]Rossi V, Bellastella G, Rosa CD, et al.Raloxifene induces cell death and inhibitsproliferation through multiple signaling pathways in prostate cancer cells.J. Cell.Physio.2011;226:1334-1339.
[48]Shen JC, Klein RD, Wei Q, et al.Low-dose genistein induces cyclin-dependentkinase inhibitors and G1cell-cycle arrest in human prostate cancer cells.MolCarcinog2000;29:92-102.
[49]Mentor-Marcel R, Lamaritiniere CA, Eltoum IE, et al.Genistein in the dietreduces the incidence of poorly differentiated prostatic adenocarcinoma intransgenic mice(TRAMP).Cancer Res.2001;61(18):6777-6782.
[50]Mak P. Leung YK, Tang WY, Harwood C, et al.Apigenin supresses cancer cellgrowth through ERbeta.Neoplasia2006;8:896-904.
[51]Skliris GP, Munot K, Bell SM, et al.Reduced expression of estrogen receptor β ininvasive breast cancer and its reexpression using DNA methyltransferaseinhibitors in a cell line mode.J Pathol2003;201:213
[52]Rody A, Holtrich U, Solbach C, et al.Methylation of estrogen receptor β promotercorrelates with loss of ERβ expression in mammary carcinoma and is an earlyindication marker in premalignant lesions.Endoc Relat Cancer2005;12:903.
[53]Zhao C, Lam EW, Sunters A, et al.Expression of estrogen receptor beta isforms innormal breast epithelial cells and breast cancer: regulaiton by methylation.Oncogene2003;22:7600-7606.
[54]Omoto Y, Inoue S, Ogawa S, et al.Clinical value of the wild-type estrogenreceptor β expression in breast cancer.Cancer Letters2001;163:207-212.
[55]Lindberg K, Strom A, Lock J G, et al.Expression of estrogen receptor β increasesintegrin α1and integrin β1levels and enhances adhesion of breast cancer cells. JCell Physiol2010;222:156-167.
[56]Lindberg et al. Estrogen receptor β represses Akt signaling in breast cancer cellsvia downregulation of HER2/HER3and upregulation of PTEN: implications fortamoxifen sensitivity. Breast Cancer Research2011;13:R43
[57]Lindgen P, Cajiander S, Backstrom T, et al.Estrogen and progesterone andprogeterone recptors in epithelial ovarian tumors.Mol Cell Endocrinol2004;22:97-104.
[58]Suzuki F, Akahira J, Miura I, et al.Loss of estrogen receptor β isoform expressionand its correlation with aberrant DNA methylation of the5’-untranslated regionin human epithelial ovarian carcinoma.Cancer Sci2008;99:2365-2372.
[59]Treeck O, Pfeiler G, Mitter D, et al.Estrogen receptor β exerts anti-tumoral effectson SK-OV-3ovarian cancer cells.Endocrinol2007;193:421.
[60]Lazennec G.Estrogen recptor beta, a possible tumor suppressor involved inovarian carcinogenesis.Cancer Lett2006;231:151-157.
[61]Campbell-Thompson M, Lynch J&Bhardwaj B.Expression of estrogenreceptor(ER) subtypes ER beta isforms in colon cancer. Cancer Research2001;61:632-640.
[62]Jassam N, Bell SM, Speirs V, et al.Loss of expression of oestrogen receptor betain colon cancer and its association with Dukes’ staging.Oncol Rep2005;14:17-21.
[63]Omoto Y, Eguchi H, Yamamoto-Yamaguchi Y,et al.Estrogen receptor(ER)β1andERβcx/β2inhibits ERα function differently in breast cancer cell lineMCF7.Oncogene2003;22:5011-5020.
[64]Paruthiyil S, Parmar H, Kerekatte V, et al.Estrogen receptor beta inhibits humanbreast cancer cell proliferation and tumor formation by causing a G2cell cyclearrest.Cancer Research2004;64:423-428.
[65]McPherson SJ, Wang H, Jones ME, et al.Elevated androgens and prolactin inaromatase-deficient mice cause enlargement, but not malignancy, of the prostategland.Endocrinology2001;142:2458-2467.
[66]Kypianou N, English HF, Davidson NE, et al.Programmed cell death duringregression of the MCF-7human breast cancer following estrogen ablation.CancerResearch1991;51:162-166.
[67]Perillo B, Sasso A, Abbondanza C, et al.17β-Estradiol inhibits apoptosis inMCF-7cells, inducing bcl-2expression via two estrogen-responsive elementspresent in the coding sequence.Molecular and Cellular Biology2000;20:2890-2901.
[68]Choi KC, Kang SK, Tai CJ, et al.Estrodiol up-regulates antiapoptotic Bcl-2messenger ribonucleic acid and protein in tumorigenic ovarian surface epitheliumcells.Endocrinology2001;142:2351-2360.
[69]Nilsen J, Mor G&Naftolin F.Estrogen-regulated developmental neuronalapoptosis is determined by estrogen receptor subtype and the Fas/Fas ligandsystem.Journal of Neurobiology2000;43:64-78.
[70]Bardin A, Hoffmann P, Boulle N, et al.Involvement of estrogen receptor β inovarian carcinogenesis. Cancer Research2004;64:5861-9.
[71]Qiu Y, Waters CE, Lewis AE, et al.Qestrogen-induced apoptosis in colonocytesexpression oestrogen receptor beta.J Endocrinol2002;174:369-77.
[72]Hsu HH, Cheng SF, Wu CC, et al.Apoptotic effects of over-expressed estrogenreceptor-beta on LoVo colon cancer cell is mediated by p53signaling in aligand-dependent manner.Chin J Physiol2006;49:110-116.
[73]Lazennec G, Bresson D, Lucas A, et al.ERβ inhibits proliferation and invasion ofbreast cancer cells.Endocrinology2001;142:4120-30.
[74]Treeck O, Pfeiler G, Mitter D, et al.Estrogen receptor β1exerts antitumoral effectson SK-OV-3ovarian cancer cells.J Endocrinol2007;193:421-433.
[75]苗丽君,王静. Akt与肿瘤的研究进展[J].国外医学.生理、病理科学与临床分册.2004;24:406-409.
[76]Milas I, Komaki R, Hachiya T, et al.Epidermal growth factor receptor,Cyelooxyenase-2, and BAX expression in the Primary nonsmall cell lung cancerand brain metastases.Clin Cancer Res2003;9:1070-1076.
[77]Liao Y, Grobholz R, Abel U, et al.Increase of AKT/PKB expression corrlates withgleason pattern in human prostate cancer.Int J Cancer2003;107:676-680.
[78]Ayala G, Thompson T, Yang G, et al.High level of phosphory-lated form of Akt-1in prostate cancer and non-neoplastic prostate tissues are strong predictors ofbiochemical recurrence. Clin Can-cer Res2004;10(4):6572-6578.
[79]Kreisberg J, Malik S, Prihoda T, et al. Phosphorylation of Akt (Ser473) is anexcellent preditor of poor clinical outcome in prostate cancer. Cancer Res2004;64(4):5232-5236.
[80]孙丽梅.乳腺癌信号转到途径中Akt激活及意义。中华肿瘤防治杂志，2006;13:1232-1234.
[81]Yamamoto S, Tomita Y, Hoshida Y, et al.Prognostic significance of activated Aktexpression in pancreatic ductal adenocarcinoma.Clin Cancer Res.2004;10:2846-2850.
[82]Balsara BR, Pei J, Mitsuuchi Y, et al.Frequent activation of AKT in non-small celllung carcinomas and preneoplastic bronchial lesions. Cancinogenesis2004;25:2053-2059.
[83]Robertson CN, Roberson KM, Padilla GM, et al.Induction of apoptosis bydithylstilbestrol in hormone-insensitive prostate cancer cells.J Natl Cancer Inst1996;88:908-917.
[84]Signoretti S, Loda M.Estrogen receptor beta in prostate cancer: Brake pedal oraccelerator? Am J Pathol2001;159:13-16.
[85]李峰等，hERβ全长基因表达载体EGFP-C1-hERβ的构建及在PC-3M细胞中的表达。《吉林大学学报（医学版）》，2006;32:189-193.
[86]李峰等,雌激素受体2对前列腺癌细胞系PC-3M增殖的影响.中国病理生理学杂志，2009;25:656-660.
[87]Iwao K, Miyoshi Y, Egawa C, et al.Quantitative analysisi of estrogen receptor-βmRNA and its variants in human breast cancers.International Jounal of Cancer2000;88:733-736.
[88]Theys J, Barbe S, Landuyt W, et al. Tumor-specific gene delivery usinggenetically engineened bacteria. Cmr Gene Ther2003;3:207-221.
[89]Ming Zhao, Meng Yang, Huaiyu Ma, et al Targeted Therapy with a SalmonellaTyphimurium Leucine-Arginine Auxotroph Cures Orthotopic Human BreastTumors in Nude Mice. Cancer Res2006;66:7647-7652.
[90]Angelakopoulos H, Hohomann EL.Pilot study of phoP/phoQ-deleted Salmonellaenterica serovar Typhimurium expressiong Helicobacter pylori urease in adultvolunteers.Infect Immun2000;68:213-41.
[91]Ling Zhang, Lifang Gao, Lijuan Zhao, et al.Intratumoral delivery and suppressionof prostate tumor growth by attenuated salmonella enterica serovar typhimuriumcarrying plasmid-based small interfering RNAs.[J].Cancer Res,2007;67(12):5859-5864.
[92]Hartman J, Lindeberg K, Morani A, et al.Estrogen receptor beta inhibitsangiogenesis and growth of T47D breast cancer xeografts.Cancer Res2006;66:11207-13.
[93]Qin XG, Hua Z, Shuang W, et al.Effects of matrine on Hep G2cell proliferationand expression of tumor relevant proteins in vitro.Pharm Biol2010;48:275-281.
[94]赖建生,夏强,张孝斌,等.增殖细胞核抗原和p27在前列腺增生和前列腺癌组织中的表达及意义.中华肿瘤杂志2004;26:476-478.
[95]Graff J, Konicek B, McNulty A, et al.Increased AKT activity contributes toprostate cancer progression by dramatically accelerating prostate tumor growthand diminishing p27Kip1expression. J Biol Chem2000;275(2):24500-24505.
[96]Sastry KS, Smith AJ, Karpova Y, et al.Diverse anti-apoptotic signaling pathwaysactivated by VIP, EGF and PI3K in prostate cancer cells converge on BAD.J BiolChem2006;281:20891-901.
[97]Giulia Pinton, Warren Thomas, Palolo Bellini, et al.Estrogen receptor exsertstumor repressive functions in human malignant pleural mesothelioma EGFRinactivation and affects response to Gefitinib.PloS ONE2010;11:e14110.
[98]Chang F, Lee JT, Navolanic PM, et al.Involement of PI3-K/Akt pathway in cellcycle progression, apoptosis, and neoplastic transformation: a target for cancerchemotherapy. Leukem ia2003;17(3):590-603.
[99]Chin YR, Toker A.Function of Akt/PKB signaling to cell motility, invasion andthe tumor stromer in cancer[J]. Cell Signaling2009;21(2):470-476.
[100]Hu W, Kavanagh JJ.Anticancer therpy targeting the apoptic pathway.LancetOncle2003;4:721-729.
[101]Cheg EH, Wei MC, Weiler S, et al.BCL-2, BCL-X(L) sequesters BH3domin-only molecules preventing BAX-and-BAK-mediated mitochondrialapoptosis. Mol Cell2001;8:70511.
[102]Wei MC, Zong WX, Cheng EH, et al.Proapoptotic BAX and BAK: a requisitegate-way to mitochondrial dysfunction and death. Science2001;292:727-30.
[103]Choi, SI, Joo SS, Yoo YM.Melationin prevents nitric oxide-induced apoptosisby increasing the interaction between14-3-3[beta] and p-Bad in SK-N-MC cells.Journal of Pineal Research2008;44(1):95-100.
[104]Gonzalez-Garcia M, Perez-Ballestro R, Ding L, et al.Bcl-x is the major Mrnafrom expressed during murine development and its product localizes to themitochondrua. Development1994;120:3033.
[105]Simoes Wust AP, Olie RA, Gautschi O, et al.Bcl-xl antisense treatment induesapoptosis in breast carcinoma cells. Int J Cancer,2000;87(4):582-590.
[106]Kirkin V, Joos S, Zorning M.The role of Bcl-2family menbers in tumorigenesis.Biochim.Biphs. Acta2004;1644:229-249.
[107]Stepien A, Izdebska M, Grzanka A.The types of cell death.Postepy Hig MedDosw2007;61:420-428.
[108]Wang ZB, Liu YQ, Cui YF.Pathways to caspase activation.Cell Biol. Int.2005;29:489-496.
[109]Zhou HL, Li XM, Meinkoth J, et al.Akt Regulates Cell Survival and Apoptosisat a Postmitochondrial Level. J Cell Biol2000;151(3):483-494.
[110]Hu W, Kavanagh JJ.Anticancer therpy targeting the apoptic pathway.LancetOncle2003;4:721-729.
[111]A Ivana Scovassa, Marc Diederich.Modulation of Poly (ADP-ribosylation) inapoptotic cells.Biochemical pharmacology2004;5:1041-1247.
[112]Coopersmith CM, Chang KC, Swanson PE, et al.Overexpression of bcl-2in theintestinal epthithelium imoroves survival in septic mice. Crit Care Med2002;30:195-201.