miRNA-214通过靶向β-catenin通路抑制肝癌生长
摘要
肝细胞癌（Hepatocellular carcinoma，HCC）是人类最常见的恶性肿瘤之一。尽管在肝癌的治疗中取得了很大进步，肝癌仍是癌症相关死亡的第三大原因，患者的5年生存率不到5％。调查肝癌发生发展的分子机制，对于开发新的诊断、治疗和预后方法具有重要意义。MicroRNA（miRNA）是一类丰富的非编码的短RNA，在转录后水平通过诱导mRNA的降解或抑制其翻译从而抑制靶基因的表达。最近，越来越多的报告表明，异常表达的miRNA与各种人类癌症，包括肝癌有关，miRNA可以作为肿瘤抑制基因或癌基因参与人类癌症的发生发展。这对肝癌的诊断和治疗应用提供了新的途径。最近的miRNA表达谱研究已经证明了miR-124在肝癌表达下调，然而，其在肝癌发生过程中的潜在的功能及相关机制仍未明确。本实验从肝癌组织标本和细胞系中检测miR-124的表达，然后构建miR-124过表达慢病毒载体在体内外研究其对肝癌生长的抑制作用，并进一步探讨其机制。
研究方法
本课题首先应用实时定量RT-PCR方法检测18对配对的肝癌组织和癌旁组织以及肝癌细胞株和正常肝细胞系LO2中成熟miR-214的表达情况。然后构建miR-214的慢病毒载体，包装慢病毒，并感染HepG2细胞，筛选慢病毒稳定株。检测稳定过表达miR-214对肝癌细胞的生长、增殖、克隆形成能力以及细胞周期的影响；并进行裸鼠成瘤实验，观察miR-214对HepG2细胞成瘤能力的影响。应用生物信息学软件预测miR-214可能作用的靶基因，并采用荧光素酶报告实验、实时定量RT-PCR、Western blot进行靶基因验证。通过β-catenin RNA干扰和恢复表达实验，检测对细胞增殖的影响。检测miR-214对β-catenin通路下游蛋白CyclinD1、c-Myc、TCF-1和LEF-1的表达水平的影响。
研究结果
1.miR-214在肝癌组织和肝癌细胞系Hep3B，SK-HEP1，SMMC-7721，Huh7，HepG2细胞中的表达显著下调。
2.建立了表达miR-214的HepG2细胞稳定株。
3.细胞生长曲线表明miR-214HepG2细胞生长缓慢；MTT实验表明miR-214过表达可抑制HepG2细胞的增殖；miR-214稳定株的HepG2细胞集落生长能力降低；细胞周期检测发现miR-214组HepG2细胞在G1/G0比例为69.28%，高于对照组的54.62%；miR-214组HepG2细胞S期细胞的比例为19.56%，明显较对照组比例33.48%低，结果提示miR-214可通过引起细胞G1期阻滞抑制肝癌细胞增殖。
4.用miR-214稳定株HepG2细胞和对照慢病毒细胞，建立裸鼠肝癌移植瘤模型，结果发现，miR-214组肿瘤生长明显慢于对照组；皮下接种BALB/c裸鼠25d后，miR-214组肿瘤体积为对照组的32%，重量为对照组的37.5%。
5.TargetScan等软件预测CTNNB1可能是miR-214作用的靶基因。荧光素酶报告实验结果表明，转染miR-214表达质粒能够抑制共转染的CTNNB1-3’-UTR载体荧光素酶基因活性，而共转染miR-214表达质粒和CTNNB1-3’-UTR靶位点突变载体，荧光素酶活性未见明显改变。Western blot检测发现miR-214可抑制β-catenin蛋白的表达；实时定量RT-PCR结果显示转染miR-214表达质粒后，β-catenin mRNA的表达量和对照组相比无显著差异。
6.转染shβ-catenin干扰质粒可降低β-catenin表达，抑制HepG2细胞增殖。对稳定表达miR-214的肝癌细胞，转染不含3'-UTR的β-catenin过表达质粒，可恢复β-catenin蛋白表达，逆转miR-214对HepG2细胞增殖的抑制作用。
7.Western blot结果显示miR-214稳定表达株中CyclinD1、c-Myc和TCF-1的表达水平下降，miR-214稳定表达株裸鼠移植肿瘤组织中CyclinD1、c-Myc和LEF-1的表达水平比对照组下降。
研究结论
1.miR-214在肝癌组织和肝癌细胞系中的表达显著下调。
2. miR-214可以在体外和在体内抑制肝癌细胞的生长。
3.CTNNB1是miR-214的靶基因。miR-214可以通过抑制β-catenin通路影响肝癌生长。
Hepatocellular carcinoma （HCC） is one of the most common malignant tumorworldwide. Despite of great progress in the treatment of HCC, it remains the thirdleading cause of cancer-related death, and the5-year survival of HCC patients is lessthan5%. Therefore, it’s very necessary to clarify the molecular mechanisms of HCCfor development of new diagnostic, treatment and prognosis strategy. MicroRNAs（miRNAs） is a class of rich non-coding short RNA inhibiting the expression oftarget genes in post-transcriptional level by inducing mRNA degradation or inhibit itstranslation. Recently, a growing number of reports indicate that abnormal expressionof miRNAs associated with a variety of human cancers, including HCC, and miRNAas a tumor suppressor or oncogene involved in the development of human cancers.This provides a new approach to the diagnosis and treatment of HCC. MicroRNAprofile in recent studies have demonstrated that miR-124is downregulated in HCC,however, its potential function in the process of HCC and related mechanisms are notyet clear.
Methods
We detected the expression lever of mature miR-214in18paired HCC tissue andadjacent tissues,and HCC cell lines including Hep3B,SK-HEP1, SMMC-7721, Huh7,HepG2cells and normal liver cells LO2by quantitative real-time RT-PCR analysis.
Lentiviral expression plasmid of miR-214（Lv-miR-214） or lentiviral control（Lv-control） and packaging systems co-transfected293T cells to produce lentivirus,then infected HepG2cells, and screened cell strains stably expressing miR-214. Cellgrowth curve and MTT assay were employed for HepG2cell proliferation assay;Colony formation assay for single tumor cell colony growth capacity assay; Flowcytometry for cell cycle analysis assay. Nude mice experiments was employed toobserve the impact of the tumorigenic ability of miR-214on HepG2cells.
Bioinformatics softwares were applicated to predict the possible target genes of miR-214, and luciferase reporter experiments, quantitative real-time RT-PCR, Westernblot for target gene validation. β-catenin RNA interference and restore expressionexperiments was employed for testing the effects of β-catenin on cell proliferation.Impact of miR-214on the expression levels of β-catenin pathway downstream proteinof CyclinD1, c-Myc,TCF-1, and LEF-1was detected by Western blot.
Results
1. miR-214is significantly downregulated in HCC tissues and HCC cell lines.
2. HepG2cell strains stably expressing miR-214were established.
3. Cell growth curves showed that HepG2cell strains stably expressing miR-214growed slowly; MTT experiments showed that miR-214overexpression inhibitsHCC cell proliferation; Colony formation experiments showed that overexpression ofmiR-214reduced tumor cell colony growth ability; The flow cytometry discoveredthe proportion of cells in G1/G0phase were54.62%in the control group, while theratio in G1/G0increased to69.28%in the experimental group; the proportion of cellsin S phase was19.56%, lower than the control group, the proportion was33.48%.This suggest that miR-214caused cell cycle G1arrest.
4. HepG2cell strains stably expressing miR-214were applicatied to establish nudemice HCC animal model, compared to the control group, the experimental grouptumors grow slowly; The volume and wight of the tumors in the experimental groupwere32%and37.5%of the control group25d after inoculated subcutaneously withBALB/c nude mice.
5.TargetScan predicted CTNNB1may be a target genes of miR-214. Luciferasereporter experimental results show that luciferase gene activity can be suppressed byco-transfection of miR-214expression plasmid and CTNNB1-3'-UTR, but not byco-transfection of miR-214expression plasmid and CTNNB1-3'-UTR-mt. Westernblot analysis showed that miR-214can inhibit the expression of β-catenin protein;qRT-PCR results showed that miR-214didn’t influrenced β-catenin mRNA expression.
6.The shβ-catenin reduceed β-catenin expression, inhibited HCC cell proliferation.Transfection of β-catenin overexpression plasmid not containing the3'-UTR ofCTNNB1restored β-catenin protein expression, reversed the inhibition of miR-214on cell proliferation.
7.Western blot showed that CyclinD1, c-Myc,LEF-1, and TCF-1expression levelsdecreased in HepG2cell strains stably expressing miR-214and nude micetransplanted tumor tissue.
Conclusions
1.miR-214is significantly downregulated in HCC tissues and HCC cell lines.
2.miR-214inhibition of the growth of HCC cells in vitro and in vivo.
3. CTNNB1is a target gene of miR-214. miR-214can affect HCC cells growth by theβ-catenin pathway.
研究方法
本课题首先应用实时定量RT-PCR方法检测18对配对的肝癌组织和癌旁组织以及肝癌细胞株和正常肝细胞系LO2中成熟miR-214的表达情况。然后构建miR-214的慢病毒载体，包装慢病毒，并感染HepG2细胞，筛选慢病毒稳定株。检测稳定过表达miR-214对肝癌细胞的生长、增殖、克隆形成能力以及细胞周期的影响；并进行裸鼠成瘤实验，观察miR-214对HepG2细胞成瘤能力的影响。应用生物信息学软件预测miR-214可能作用的靶基因，并采用荧光素酶报告实验、实时定量RT-PCR、Western blot进行靶基因验证。通过β-catenin RNA干扰和恢复表达实验，检测对细胞增殖的影响。检测miR-214对β-catenin通路下游蛋白CyclinD1、c-Myc、TCF-1和LEF-1的表达水平的影响。
研究结果
1.miR-214在肝癌组织和肝癌细胞系Hep3B，SK-HEP1，SMMC-7721，Huh7，HepG2细胞中的表达显著下调。
2.建立了表达miR-214的HepG2细胞稳定株。
3.细胞生长曲线表明miR-214HepG2细胞生长缓慢；MTT实验表明miR-214过表达可抑制HepG2细胞的增殖；miR-214稳定株的HepG2细胞集落生长能力降低；细胞周期检测发现miR-214组HepG2细胞在G1/G0比例为69.28%，高于对照组的54.62%；miR-214组HepG2细胞S期细胞的比例为19.56%，明显较对照组比例33.48%低，结果提示miR-214可通过引起细胞G1期阻滞抑制肝癌细胞增殖。
4.用miR-214稳定株HepG2细胞和对照慢病毒细胞，建立裸鼠肝癌移植瘤模型，结果发现，miR-214组肿瘤生长明显慢于对照组；皮下接种BALB/c裸鼠25d后，miR-214组肿瘤体积为对照组的32%，重量为对照组的37.5%。
5.TargetScan等软件预测CTNNB1可能是miR-214作用的靶基因。荧光素酶报告实验结果表明，转染miR-214表达质粒能够抑制共转染的CTNNB1-3’-UTR载体荧光素酶基因活性，而共转染miR-214表达质粒和CTNNB1-3’-UTR靶位点突变载体，荧光素酶活性未见明显改变。Western blot检测发现miR-214可抑制β-catenin蛋白的表达；实时定量RT-PCR结果显示转染miR-214表达质粒后，β-catenin mRNA的表达量和对照组相比无显著差异。
6.转染shβ-catenin干扰质粒可降低β-catenin表达，抑制HepG2细胞增殖。对稳定表达miR-214的肝癌细胞，转染不含3'-UTR的β-catenin过表达质粒，可恢复β-catenin蛋白表达，逆转miR-214对HepG2细胞增殖的抑制作用。
7.Western blot结果显示miR-214稳定表达株中CyclinD1、c-Myc和TCF-1的表达水平下降，miR-214稳定表达株裸鼠移植肿瘤组织中CyclinD1、c-Myc和LEF-1的表达水平比对照组下降。
研究结论
1.miR-214在肝癌组织和肝癌细胞系中的表达显著下调。
2. miR-214可以在体外和在体内抑制肝癌细胞的生长。
3.CTNNB1是miR-214的靶基因。miR-214可以通过抑制β-catenin通路影响肝癌生长。
Hepatocellular carcinoma （HCC） is one of the most common malignant tumorworldwide. Despite of great progress in the treatment of HCC, it remains the thirdleading cause of cancer-related death, and the5-year survival of HCC patients is lessthan5%. Therefore, it’s very necessary to clarify the molecular mechanisms of HCCfor development of new diagnostic, treatment and prognosis strategy. MicroRNAs（miRNAs） is a class of rich non-coding short RNA inhibiting the expression oftarget genes in post-transcriptional level by inducing mRNA degradation or inhibit itstranslation. Recently, a growing number of reports indicate that abnormal expressionof miRNAs associated with a variety of human cancers, including HCC, and miRNAas a tumor suppressor or oncogene involved in the development of human cancers.This provides a new approach to the diagnosis and treatment of HCC. MicroRNAprofile in recent studies have demonstrated that miR-124is downregulated in HCC,however, its potential function in the process of HCC and related mechanisms are notyet clear.
Methods
We detected the expression lever of mature miR-214in18paired HCC tissue andadjacent tissues,and HCC cell lines including Hep3B,SK-HEP1, SMMC-7721, Huh7,HepG2cells and normal liver cells LO2by quantitative real-time RT-PCR analysis.
Lentiviral expression plasmid of miR-214（Lv-miR-214） or lentiviral control（Lv-control） and packaging systems co-transfected293T cells to produce lentivirus,then infected HepG2cells, and screened cell strains stably expressing miR-214. Cellgrowth curve and MTT assay were employed for HepG2cell proliferation assay;Colony formation assay for single tumor cell colony growth capacity assay; Flowcytometry for cell cycle analysis assay. Nude mice experiments was employed toobserve the impact of the tumorigenic ability of miR-214on HepG2cells.
Bioinformatics softwares were applicated to predict the possible target genes of miR-214, and luciferase reporter experiments, quantitative real-time RT-PCR, Westernblot for target gene validation. β-catenin RNA interference and restore expressionexperiments was employed for testing the effects of β-catenin on cell proliferation.Impact of miR-214on the expression levels of β-catenin pathway downstream proteinof CyclinD1, c-Myc,TCF-1, and LEF-1was detected by Western blot.
Results
1. miR-214is significantly downregulated in HCC tissues and HCC cell lines.
2. HepG2cell strains stably expressing miR-214were established.
3. Cell growth curves showed that HepG2cell strains stably expressing miR-214growed slowly; MTT experiments showed that miR-214overexpression inhibitsHCC cell proliferation; Colony formation experiments showed that overexpression ofmiR-214reduced tumor cell colony growth ability; The flow cytometry discoveredthe proportion of cells in G1/G0phase were54.62%in the control group, while theratio in G1/G0increased to69.28%in the experimental group; the proportion of cellsin S phase was19.56%, lower than the control group, the proportion was33.48%.This suggest that miR-214caused cell cycle G1arrest.
4. HepG2cell strains stably expressing miR-214were applicatied to establish nudemice HCC animal model, compared to the control group, the experimental grouptumors grow slowly; The volume and wight of the tumors in the experimental groupwere32%and37.5%of the control group25d after inoculated subcutaneously withBALB/c nude mice.
5.TargetScan predicted CTNNB1may be a target genes of miR-214. Luciferasereporter experimental results show that luciferase gene activity can be suppressed byco-transfection of miR-214expression plasmid and CTNNB1-3'-UTR, but not byco-transfection of miR-214expression plasmid and CTNNB1-3'-UTR-mt. Westernblot analysis showed that miR-214can inhibit the expression of β-catenin protein;qRT-PCR results showed that miR-214didn’t influrenced β-catenin mRNA expression.
6.The shβ-catenin reduceed β-catenin expression, inhibited HCC cell proliferation.Transfection of β-catenin overexpression plasmid not containing the3'-UTR ofCTNNB1restored β-catenin protein expression, reversed the inhibition of miR-214on cell proliferation.
7.Western blot showed that CyclinD1, c-Myc,LEF-1, and TCF-1expression levelsdecreased in HepG2cell strains stably expressing miR-214and nude micetransplanted tumor tissue.
Conclusions
1.miR-214is significantly downregulated in HCC tissues and HCC cell lines.
2.miR-214inhibition of the growth of HCC cells in vitro and in vivo.
3. CTNNB1is a target gene of miR-214. miR-214can affect HCC cells growth by theβ-catenin pathway.
引文
[1]Kamangar F, Dores GM, Anderson WF. Patterns of cancer incidence,mortality,andprevalence across five continents:defining priorities to reduce cancer disparities indifferent geographic regions of the world[J].J Clin Oncol,2006,24(14):2137–2150.
[2] Parkin DM, Bray F, Ferlay J, et al. Global cancer statistics,2002[J]. CA Cancer JClin,2005,55(2):74-108.
[3]Kumar MS, Lu J, Mercer KL, et al. Impaired microRNA processing enhancescellular transformation and tumorigenesis[J]. Nat Genet,2007,39(5):673-677.
[4]Pillai RS. MicroRNA function:multiple mechanisms for a tiny RNA?[J].RNA2005,11(12):1753-1761.
[5]Shenouda SK,Alahari SK. MicroRNA function in cancer:oncogene or a tumorsuppressor?[J]. Cancer Metastasis Rev,2009,28(3-4):369-378.
[6]Ueda R, Kohanbash G, Sasaki K,et al. Dicer-regulated microRNAs222and339promote resistance of cancer cells to cytotoxic T-lymphocytes by down-regulationof ICAM-1[J].Proc Natl Acad Sci U S A,2009,106(26):10746-10751.
[7]Ma L,Teruya-Feldstein J,Weinberg RA. Tumour invasion and metastasis initiatedby microRNA-10b in breast cancer.Nature,2007,449(7163):682-688.
[8]Varnholt H.The role of microRNAs in primary liver cancer[J].Annals ofHepatology,2008,7(2):104-113.
[9]Meng F, Henson R, Wehbe-Janek H, et a1. MicroRNA-21regulates expression ofthe PTEN tumor suppressor gene in human hepatocellular cancer[J].Gastroenterology,2007,133(2):647-658.
[10]Ma L,Liu J,Shen J,et a1．Expression of miR-122mediated by adenoviral vectorinduces apoptosis and cell cycle arrest of cancer cells[J]． Cancer BiolTher,2010,9(7):554-561.
[11]Tsai W C, Hsu PW, Lai TC,et a1. MicroRNA-122，a tumor suppressor microRNAthat regulates intrahepatic metastasis of hepatocellu1ar carcinoma[J]. Hepatology，2009,49(5):1571-1582.
[12]Bai S, Nasser MW, Wang B, et a1. MicroRNA-122inhibits tumorigenicproperties of hepatocellular carcinoma cells and sensitizes these cells tosorafenib[J]. J Biol Chem,2009,284(46)：32015-32027．
[13]Fornari F, Milazzo M, Chieco P, et a1. MiR-199a-3p regulates mTOR and c-Metto influence the doxorubicin sensitivity of human hepatocarcinoma cells[J].Cancer Res,2010,70(12)：5184-5193．
[14]Wong CC, Wong CM, Tung EK, et al. The microRNA miR-139suppressesmetastasis and progression of hepatocellular carcinoma by down-regulatingRho-kinase2[J]. Gastroenterology,2011,140(1):322-331.
[15] Wang Y, Lee AT, Ma JZ, et al. Profiling microRNA expression in hepatocellularcarcinoma reveals microRNA-224up-regulation and apoptosis inhibitor-5as amicroRNA-224-specific target[J]. J Biol Chem,2008,283(19):13205-13215.
[16] Yang Z, Chen S, Luan X, et al. MicroRNA-214is aberrantly expressed incervical cancers and inhibits the growth of HeLa cells[J]. IUBMB Life,2009,61:1075-1082.
[17] Derfoul A, Juan AH, Difilippantonio MJ, et al. Decreased microRNA-214levelsin breast cancer cells coincides with increased cell proliferation, invasion andaccumulation of the Polycomb Ezh2methyltransferase[J]. Carcinogenesis,2011,32:1607-1614.
[18]Zhang XJ, Ye H, Zeng CW, et al. Dysregulation of miR-15a and miR-214inhuman pancreatic cancer[J]. Journal of hematology&oncology,2010,3:46.
[19]Yang H, Kong W, He L, et al. MicroRNA expression profiling in human ovariancancer: miR-214induces cell survival and cisplatin resistance by targetingPTEN[J]. Cancer Res,2008,68(2):425–433.
[20]Thompson MD,Monga SP. WNT/beta-catenin signaling in liver health anddisease [J]. Hepatology,2007,45:1298-305.
[21]Clevers H.Wnt/beta-catenin signaling in development and disease[J].Cell.2006,127:469-480.
[22]Cai SR, Wang Z, Chen CQ, et al. Role of silencing phosphatase of regenerationgliver-3expression by microRNA interference in the growth of gastriccancer[J].Chin Med J (Engl),2008,121(24):2534-2538.
[23]Tazawa H, Tsuchiya N, Izumiya M, et al. Tumor-suppressive miR-34a inducessenescence-like growth arrest through modulation of the E2F pathway in humancolon cancer cells[J]. Proc Natl Acad Sci,2007,104(39):15472-15477.
[24] Rossi JJ. New hope for a microRNA therapy for liver cancer[J]. Cell,2009,137(6):990-992.
[25]Kota J, Chivukula RR, O'Donnell KA, et al. Therapeutic miRNA deliverysuppresses tumorigenesis in a murine liver cancer model [J]. Cell,2009,137(6):1005-1017.
[26]张敬磊,金华君,刘辉,等.腺病毒介导microRNA-99a过表达抑制肝癌细胞生长[J].中国肿瘤生物治疗杂志,2012,19(3):267-271.
[27] Connolly E, Melegari M, Landgraf P, et al.Elevated expression of the miR-17-92polycistron and miR-21in hepadnavirus-associated hepatocellular carcinomacontributes to the malignant phenotype[J]. Am J Pathol.2008,173(3):856-864.
[28]Xu XR, Huang J, Xu ZG, et al. Insight into hepatocellular carcinogenesis attranscriptome level by comparing gene expression profiles of hepatocellularcarcinoma with those of corresponding noncancerous liver[J]. Proc Natl Acad SciUSA,2001,98(26):15089-15094.
[29]Satoh S, Daigo Y, Furukawa Y, et al. AXIN1mutations in hepatocellularcarcinomas, and growth suppression in cancer cells by virus-mediated transfer ofAXIN1[J]. Nat Genet,2000,24(3):245-250.
[30]Ding Q, Xia W, Liu JC, et al. Erk associates with and primes GSK-3beta for itsinactivation resulting in upregulation of beta-catenin[J]. Mol Cell,2005,19(2):159-170.
[31] Lian Z, Liu J, Li L, et al. Enhanced cell survival of Hep3B cells by the hepatitis Bx antigen effector, URG11, is associated with upregulation of beta-catenin[J].Hepatology,2006,43(3):415-424.
[32]Murakami H, Sanderson ND, Nagy P, et al.Transgenic mouse model forsynergistic effects of nuclear oncogenes and growth factors in tumorigenesis:interaction of c-myc and transforming growth factor alpha in hepaticoncogenesis[J]. Cancer Res,1993,53(8):1719-1723.
[33]Shachaf CM, Kopelman AM, Arvanitis C, et al. MYC inactivation uncoverspluripotent differentiation and tumour dormancy in hepatocellular cancer[J].Nature,2004,431(7012):1112-1117
[34]Dowdy SF, Hinds PW, Louie K, et al. Physical interaction of the retinoblastomaprotein with human D cyclins[J].Cell,1993,73(3):499-511.
[35]Jiang W, Zhang YJ, Kahn SM, et al. Altered expression of the cyclin D1andretinoblastoma genes in human esophageal cancer[J]. PNAS,1993,90(19):9026-9030.
[36]Nishida N, Fukuda Y, Komeda T, et al. Amplification and overexpression ofthecyclin D1gene in aggressive human hepatocellular carcinoma[J]. Caner Res.1994,54(12):3107-3110.
[37]Wong CM, Ng IO. Molecular pathogenesis of hepatocellular carcinoma[J].Liver Int2008,28(2):160–174.
[38]Polakis P. Wnt signaling and cancer[J].Genes Dev,2000,14(15):1837–1851.
[39]MacDonald BT, Tamai K, He X. Wnt/beta-catenin signaling: components,mechanisms, and diseases[J].Dev Cell,2009,17(1):9–26.
[40]Chen JP, Lin C, Xu CP, et al. Molecular therapy with recombinant antisensec-myc adenovirus for human gastric carcinoma cells in vitro and in vivo[J]. JGastroenterol Hepatol,2001,16(1):22–28.
[41]Wong NA, Morris RG, McCondochie A, et al. Cyclin D1overexpression incolorectal carcinoma in vivo is dependent on beta-catenin protein dysregulation,but not k-ras mutation[J].J Pathol2002;197(1):128–135.
[42]Huang K, Zhang JX, Han L, et al. MicroRNA roles in beta-catenin pathway[J].Mol Cancer,2010,9:252.
[43] Hashimi ST, Fulcher JA, Chang MH, et al. MicroRNA profiling identifiesmiR-34a and miR-21and their target genes JAG1and WNT1in the coordinateregulation of dendritic cell differentiation[J]. Blood,2009,114(2):404–414.
[44]Burk U, Schubert J, Wellner U, et al. A reciprocal repression between ZEB1andmembers of the miR-200family promotes EMT and invasion in cancer cells[J].EMBO Rep,2008,9(6):582–589.
[45]Su J, Zhang A, Shi Z, et al. MicroRNA-200a suppresses the Wnt/β-cateninsignaling pathway by interacting with β-catenin[J]. Int J Oncol.2012,40(4):1162-1170.
[46]Xu J, Zhu X, Wu L, et al. MicroRNA-122suppresses cell proliferation andinduces cell apoptosis in hepatocellular carcinoma by directly targetingWnt/β-catenin pathway[J]. Liver Int,2012,32(5):752-760.
[47] Huang SD, Yuan Y, Zhuang CW, et al. MicroRNA-98and microRNA-214post-transcriptionally regulate enhancer of zeste homolog2and inhibit migrationand invasion in human esophageal squamous cell carcinoma[J]. Mol Cancer,2012,11:51.
[48]Zhao Z, Tan X, Zhao A, et al. microRNA-214-mediated UBC9expression inglioma[J]. BMB Rep,2012,45(11):641-646.
[49] Ueda T, Volinia S, Okumura H, et al. Relation between microRNA expressionand progression and prognosis of gastric cancer: a microRNA expressionanalysis[J]. Lancet Oncol,2010,11(2):136–146.
[50] Penna E, Orso F, Cimino D, et al. MicroRNA-214contributes to melanomatumour progression through suppression of TFAP2C[J]. EMBO J,2011,30(10):1990–2007.
[1] Chen Y, Shen A, Rider PJ, et al. A liver-specific microRNA binds to a highlyconserved RNA sequence of hepatitis B virus and negatively regulates viral geneexpression and replication [J]. FASEB J,2011,25(12):4511-4521.
[2] Zhang Y, Jia Y, Zheng R, et al. Plasma microRNA-122as a biomarker for viral-,alcohol-and chemical-related hepatic diseases [J]. Clin Chem,2010,56(12):1830-1838.
[3]贾音,张毅,费明钰,等.慢性乙肝患者外周血microRNAs的表达变化[J].第二军医大学学报,2010,31(12):1381-1383.
[4] Gui J, Tian Y, Wen X, et al. Serum microRNA characterization identifiesmiR-885-5p as a potential marker for detecting liver pathologies [J]. Clin Sci(Lond).,2011,120(5):183-193.
[5] Xu J, Wu C, Chen X, et al. Circulating microRNAs, miR-21, miR-122, andmiR-223, in patients with hepatocellular carcinoma or chronic hepatitis [J]. MolCarcinog,2011,50(2):136-142.
[6] Ji F, Yang B, Peng X, et al. Circulating microRNAs in hepatitis B virus-infectedpatients[J]. J Viral Hepat,2011,18(7): e242-251.
[7]张静,顾国浩,史进方,等.重型乙型肝炎患者外周血单个核细胞microRNA表达及其意义[J].临床检验杂志,2012,30(3):219-221.
[8] Wu FL, Jin WB, Li JH, et al. Targets for human encoded microRNAs in HBVgenes [J]. Virus Genes,2011,42(2):157-161.
[9] Zhang GL, Li YX, Zheng SQ, et al. Suppression of hepatitis B virus replicationby microRNA-199a-3p and microRNA-210[J]. Antiviral Res,2010,88(2):169-175.
[10]Zhang X, Zhang E, Ma Z, et al. Modulation of hepatitis B virus replication andhepatocyte differentiation by MircoRNA-1[J]. Hepatology,2011,53(5):1476-1485.
[11]郜玉峰,余莉,李家斌,等.靶向ASGPR1的外源性microRNA对HBV表达和复制的抑制作用[J].世界华人消化杂志,2009,17(7):699-704.
[12]李宁,陈明泉,程琦,等.调控慢性乙型肝炎患者I型干扰素信号通路的microRNA的筛选及鉴定[J].中国病毒病杂志,2012,2(2):107-112.
[13]Randall G, Panis M, Cooper JD, et al. Celluar cofactors affecting hepatitis C virusinfection and replication [J]. Proc Natl Acad Sci U S A,2007,104(31):12884-12889.
[14]Narbus CM, Israelow B, Sourisseau M, et al. HepG2cells expressing microRNAmiR-122support the entire hepatitis C virus life cycle [J]. J Virol,2011,85(22):12087-12092.
[15]Bihrer V, Friedrich-Rust M, Kronenberger B, et al. Serum miR-122as abiomarker of necroinflammation in patients with chronic hepatitis C virusinfection [J]. Am J Gastroenterol,2011,106(9):1663-1669.
[16]Sarason FM, Krol J, Markiewicz I, et al. Decreased levels of microRNA-122inindividuals with hepatitis C responding poorly to interferon therapy [J]. Nat Med,2009,15(1):31-33.
[17]Pedersen IM, Cheng G, Wieland S, et al. Interferon modulation of cellularmicroRNAs as an antiviral mechanism [J]. Nature,2007,449(7164):919-922.
[18]Scagnolari C, Zingariello P, Vecchiet J, et al. Differential expression ofinterferon-induced microRNAs in patients with chronic hepatitis C virus infectiontreated with pegylated interferon alpha [J]. Virol J,2010,(7):311.
[19]胡学玲. microRNA在乙型、丙型肝炎中的作用[J].临床肝胆病杂志,2012,28(11):877-880.
[20]Zhang Y, Jia Y, Zheng R, et al. Plasma microRNA-122as a biomarker for viral-,alcohol-, and chemical-related hepatic diseases [J]. Clin Chem,2010,56(12):1830-8.
[21]Wang K, Zhang S, Marzolf B, et al. Circulating microRNAs, potential biomarkersfor drug-induced liver injury [J]. Proc Natl Acad Sci USA,2009,106(11):4402-4407.
[22]Murakami Y, Toyoda H, Tanaka M, et al. The progression of liver fibrosis isrelated with overexpression of the miR-199and200families [J]. PLoS One,2011,6(1):e16081
[23]Roderburg C, Urban GW, Bettermann K, et al. Micro-RNA profiling reveals arole for miR-29in human and murine liver fibrosis [J]. Hepatology,2011,53(1):209-218.
[24]Li WQ, Chen C, Xu MD, et al. The rno-miR-34family is upregulated and targetsACSL1in dimethylnitrosamine-induced hepatic fibrosis in rats [J]. FEBS J.2011,278(9):1522-1532.
[25]Chen C, Wu CQ, Zhang ZQ, et al. Loss of expression of miR-335is implicated inhepatic stellate cell migration and activation. Exp Cell Res.2011,317(12):1714-1725.
[26]Furuta M, Ozawkie KI, Kanaka S, et al. miRNA-124and miRNA-203areepigenetically silenced tumor-suppressive Microfloras in hepatocellularcarcinoma [J]. Carcinogenesis,2010,31(5):766-776.
[27]Ivanovska I, Ball AS, Diaz RL, et al. MicroRNAs in the miR-106b familyregulate p21/CDKN1A and promote cell cycle progression[J]. Mol Cell Biol,2008,28(7):2167-2174.
[28]Fornent F, Lazzo M, Chico P, et al. miRNA-199a-3p regulates mTor and c-Met toinfluence the doxorubicin sensitivity of human adenocarcinomata cells[J]. CancerRes,2010,70(12):5184-5193.
[29]Hou J, Lin L, Zhou W, et al. Identification of miRNomes in human liver andhepatocellular carcinoma reveals miR-199a/b-3p as therapeutic target forhepatocellular carcinoma [J]. Cancer Cell,2011,19(2):232-243.
[30]Jia XQ, Cheng HQ, Qian X, et al. Lentivirus-mediated overexpression ofmicroRNA-199a inhibits cell proliferation of human hepatocellular carcinoma.[J].Cell Biochem Biophys,2012,62(1):237-244.
[31]Henry JC, Park JK, Jiang J, et al. MiR-199a-3p targets CD44and reducesproliferation of CD44positive hepatocellular carcinoma cell lines [J]. BiochemBiophys Res Commun,2010,403:120-125.
[32]Buurman R, Gurlevik E, Schaffer V, et al. Histone deacetylases activatehepatocyte growth factor signaling by repressing microRNA-449inhepatocellular carcinoma cells [J]. Gastroenterology,2012,[Epub ahead ofprint].
[33]Xu J, Zhu X, Wu L, et al. MicroRNA-122suppresses cell proliferation andinduces cell apoptosis in hepatocellular carcinoma by directly targetingWnt/β-catenin pathway [J]. Liver Int,2012,32(5):752-760.
[34]Chung GE, Mung SJ, Lee SH, et al. High expression of microRNA-15b predicts alow risk of tumor recurrence following curative resection of hepatocellularcarcinoma [J]. Colon Report,2010,23(1):113-119.
[35]Guo CJ, Pan Q, Li DG, et al. miR-15b and miR-16are implicated in activation ofthe rat hepatic stellate cell: An essential role for apoptosis [J]. J Hepatol,2009,50(4):766-778.
[36]张俊,罗娜,王勃,等.上调microRNA-150表达对肝癌SMMC7721细胞增殖和凋亡的影响[J].解放军医学杂志,2012,37(10):943-936.
[37]Li D, Liu X, Lin L, et al. MicroRNA-99a inhibits hepatocellular carcinomagrowth and correlates with prognosis of patients with hepatocellular carcinoma[J]. J Biol Chem,2011,286(42):36677-36685.
[38]张敬磊,金华君,刘辉,等.腺病毒介导microRNA-99a过表达抑制肝癌细胞生长[J].中国肿瘤生物治疗杂志,2012,19(3):267-271.
[39]Nohata N, Hanazawa T, Kikkawa N, et al. Tumor suppressive microRNA-375regulates oncogene AEG-1/MTDH in head and neck squamous cell carcinoma(HNSCC)[J]. J Human Genetics,2011,56(8):595-601.
[40]朱秀明. MicroRNAlet-7c在肝细胞癌中的表达、功能及靶点的研究[D].浙江大学,2012.
[41]Yao J, Liang L, Huang S, et al. MicroRNA-30d promotes tumor invasion andmetastasis by targeting Galphai2in hepatocellular carcinoma [J]. Hepatology,2010,51(3):846-856.
[42]Ding J, Huang S, Wu S, et al. Gain of miR-151on chromosome8q24.3facilitatestumour cell migration and spreading through downregulating RhoGDIA [J]. NatCell Biol,2010,12(4):390-399.
[43]Tsai WC, Lai TC, Chau GY, et al. MicroRNA-122，a tumor suppressor microRNAthat regulates intrahepatic metastasis of hepatocellular carcinoma [J]. Hepatology,2009,49(5):1571-1582.
[44]肖海静,王观宇,董庆华.人肝细胞肝癌和癌旁正常组织microRNA表达差异的分析[J].肿瘤防治研究,2012,39(8):947-949.
[45]Sukata T, Sumida K, Kushida M, et al. Circulating miRNAs, possible indicatorsof progress of rat hepatocarcinogenesis from early stages [J]. Toxicol Lett,2011,200(1-2):46-52.
[46]高军平,向邦德,倪航航,等. microRNA-224在肝细胞癌组织和血浆中表达的相关性[J].中国现代医学杂志,2011,32(21):4012-4021.
[47]Li LM, Hu ZB, Zhou ZX, et al. Serum miRNA profiles serve asnovel biomarkersfor HBV infection and diagnosis of HBV-positive hepatocarcinoma[J]. CancerRes,2010,70(23):9798－9807.
[48]Kota J, Chivukula RR, O'Donnell KA, et al. Therapeutic miRNA deliverysuppresses tumorigenesis in a murine liver cancer model [J]. Cell,2009,137(6):1005-1017.
[49]Connolly E, Melegari M, Landgraf P, et al.Elevated expression of the miR-17-92polycistron and miR-21in hepadnavirus-associated hepatocellular carcinomacontributes to the malignant phenotype[J]. Am J Pathol.2008,173(3):856-864.
[50]Zhou C, Liu J, Li Y, et al. microRNA-1274a, a modulator of sorafenib induced adisintegrin and metalloproteinase9(ADAM9) down-regulation in hepatocellularcarcinoma [J]. FEBS Lett,2011,585:1828-1834.
[51]Li J, Wang Y, Yu W, et al. Expression of yserum miRNA-221in humanhepatocellular carcinoma and its prognostic significance [J]. Biochem BiophysRes Commun,2011,406(1):70-73.
[52]Wang C, Song B, Song W, et al. Unpressed miRNA-199b-5p targetsHypoxia-lnducible Factor-1a in hepatocellular carcinoma and predicts prognosisof hepatocellular carcinoma patients [J]. J Gastroenteroi Hepatol,2011,26(11):1630-1637.
[53]Ji JF, Shi J, Budhu A, et al. miRNA expression, survival, and response tointerferon in liver cancer [J]. N Engl J Med,2009,361(15):1437-1447.
[54]倪航航,向邦德,高军平,等.血浆中microRNA-122与肝癌手术肝损伤的相关性研究[J].实用医学杂志,2012,28(6):932-935.
[55]邓治亮,孙建,区应亮,等.原发性肝癌术后早期复发密切相关microRNA的筛选及应用[J].中山大学学报(医学科学版),2012,33(4):494-498.
[2] Parkin DM, Bray F, Ferlay J, et al. Global cancer statistics,2002[J]. CA Cancer JClin,2005,55(2):74-108.
[3]Kumar MS, Lu J, Mercer KL, et al. Impaired microRNA processing enhancescellular transformation and tumorigenesis[J]. Nat Genet,2007,39(5):673-677.
[4]Pillai RS. MicroRNA function:multiple mechanisms for a tiny RNA?[J].RNA2005,11(12):1753-1761.
[5]Shenouda SK,Alahari SK. MicroRNA function in cancer:oncogene or a tumorsuppressor?[J]. Cancer Metastasis Rev,2009,28(3-4):369-378.
[6]Ueda R, Kohanbash G, Sasaki K,et al. Dicer-regulated microRNAs222and339promote resistance of cancer cells to cytotoxic T-lymphocytes by down-regulationof ICAM-1[J].Proc Natl Acad Sci U S A,2009,106(26):10746-10751.
[7]Ma L,Teruya-Feldstein J,Weinberg RA. Tumour invasion and metastasis initiatedby microRNA-10b in breast cancer.Nature,2007,449(7163):682-688.
[8]Varnholt H.The role of microRNAs in primary liver cancer[J].Annals ofHepatology,2008,7(2):104-113.
[9]Meng F, Henson R, Wehbe-Janek H, et a1. MicroRNA-21regulates expression ofthe PTEN tumor suppressor gene in human hepatocellular cancer[J].Gastroenterology,2007,133(2):647-658.
[10]Ma L,Liu J,Shen J,et a1．Expression of miR-122mediated by adenoviral vectorinduces apoptosis and cell cycle arrest of cancer cells[J]． Cancer BiolTher,2010,9(7):554-561.
[11]Tsai W C, Hsu PW, Lai TC,et a1. MicroRNA-122，a tumor suppressor microRNAthat regulates intrahepatic metastasis of hepatocellu1ar carcinoma[J]. Hepatology，2009,49(5):1571-1582.
[12]Bai S, Nasser MW, Wang B, et a1. MicroRNA-122inhibits tumorigenicproperties of hepatocellular carcinoma cells and sensitizes these cells tosorafenib[J]. J Biol Chem,2009,284(46)：32015-32027．
[13]Fornari F, Milazzo M, Chieco P, et a1. MiR-199a-3p regulates mTOR and c-Metto influence the doxorubicin sensitivity of human hepatocarcinoma cells[J].Cancer Res,2010,70(12)：5184-5193．
[14]Wong CC, Wong CM, Tung EK, et al. The microRNA miR-139suppressesmetastasis and progression of hepatocellular carcinoma by down-regulatingRho-kinase2[J]. Gastroenterology,2011,140(1):322-331.
[15] Wang Y, Lee AT, Ma JZ, et al. Profiling microRNA expression in hepatocellularcarcinoma reveals microRNA-224up-regulation and apoptosis inhibitor-5as amicroRNA-224-specific target[J]. J Biol Chem,2008,283(19):13205-13215.
[16] Yang Z, Chen S, Luan X, et al. MicroRNA-214is aberrantly expressed incervical cancers and inhibits the growth of HeLa cells[J]. IUBMB Life,2009,61:1075-1082.
[17] Derfoul A, Juan AH, Difilippantonio MJ, et al. Decreased microRNA-214levelsin breast cancer cells coincides with increased cell proliferation, invasion andaccumulation of the Polycomb Ezh2methyltransferase[J]. Carcinogenesis,2011,32:1607-1614.
[18]Zhang XJ, Ye H, Zeng CW, et al. Dysregulation of miR-15a and miR-214inhuman pancreatic cancer[J]. Journal of hematology&oncology,2010,3:46.
[19]Yang H, Kong W, He L, et al. MicroRNA expression profiling in human ovariancancer: miR-214induces cell survival and cisplatin resistance by targetingPTEN[J]. Cancer Res,2008,68(2):425–433.
[20]Thompson MD,Monga SP. WNT/beta-catenin signaling in liver health anddisease [J]. Hepatology,2007,45:1298-305.
[21]Clevers H.Wnt/beta-catenin signaling in development and disease[J].Cell.2006,127:469-480.
[22]Cai SR, Wang Z, Chen CQ, et al. Role of silencing phosphatase of regenerationgliver-3expression by microRNA interference in the growth of gastriccancer[J].Chin Med J (Engl),2008,121(24):2534-2538.
[23]Tazawa H, Tsuchiya N, Izumiya M, et al. Tumor-suppressive miR-34a inducessenescence-like growth arrest through modulation of the E2F pathway in humancolon cancer cells[J]. Proc Natl Acad Sci,2007,104(39):15472-15477.
[24] Rossi JJ. New hope for a microRNA therapy for liver cancer[J]. Cell,2009,137(6):990-992.
[25]Kota J, Chivukula RR, O'Donnell KA, et al. Therapeutic miRNA deliverysuppresses tumorigenesis in a murine liver cancer model [J]. Cell,2009,137(6):1005-1017.
[26]张敬磊,金华君,刘辉,等.腺病毒介导microRNA-99a过表达抑制肝癌细胞生长[J].中国肿瘤生物治疗杂志,2012,19(3):267-271.
[27] Connolly E, Melegari M, Landgraf P, et al.Elevated expression of the miR-17-92polycistron and miR-21in hepadnavirus-associated hepatocellular carcinomacontributes to the malignant phenotype[J]. Am J Pathol.2008,173(3):856-864.
[28]Xu XR, Huang J, Xu ZG, et al. Insight into hepatocellular carcinogenesis attranscriptome level by comparing gene expression profiles of hepatocellularcarcinoma with those of corresponding noncancerous liver[J]. Proc Natl Acad SciUSA,2001,98(26):15089-15094.
[29]Satoh S, Daigo Y, Furukawa Y, et al. AXIN1mutations in hepatocellularcarcinomas, and growth suppression in cancer cells by virus-mediated transfer ofAXIN1[J]. Nat Genet,2000,24(3):245-250.
[30]Ding Q, Xia W, Liu JC, et al. Erk associates with and primes GSK-3beta for itsinactivation resulting in upregulation of beta-catenin[J]. Mol Cell,2005,19(2):159-170.
[31] Lian Z, Liu J, Li L, et al. Enhanced cell survival of Hep3B cells by the hepatitis Bx antigen effector, URG11, is associated with upregulation of beta-catenin[J].Hepatology,2006,43(3):415-424.
[32]Murakami H, Sanderson ND, Nagy P, et al.Transgenic mouse model forsynergistic effects of nuclear oncogenes and growth factors in tumorigenesis:interaction of c-myc and transforming growth factor alpha in hepaticoncogenesis[J]. Cancer Res,1993,53(8):1719-1723.
[33]Shachaf CM, Kopelman AM, Arvanitis C, et al. MYC inactivation uncoverspluripotent differentiation and tumour dormancy in hepatocellular cancer[J].Nature,2004,431(7012):1112-1117
[34]Dowdy SF, Hinds PW, Louie K, et al. Physical interaction of the retinoblastomaprotein with human D cyclins[J].Cell,1993,73(3):499-511.
[35]Jiang W, Zhang YJ, Kahn SM, et al. Altered expression of the cyclin D1andretinoblastoma genes in human esophageal cancer[J]. PNAS,1993,90(19):9026-9030.
[36]Nishida N, Fukuda Y, Komeda T, et al. Amplification and overexpression ofthecyclin D1gene in aggressive human hepatocellular carcinoma[J]. Caner Res.1994,54(12):3107-3110.
[37]Wong CM, Ng IO. Molecular pathogenesis of hepatocellular carcinoma[J].Liver Int2008,28(2):160–174.
[38]Polakis P. Wnt signaling and cancer[J].Genes Dev,2000,14(15):1837–1851.
[39]MacDonald BT, Tamai K, He X. Wnt/beta-catenin signaling: components,mechanisms, and diseases[J].Dev Cell,2009,17(1):9–26.
[40]Chen JP, Lin C, Xu CP, et al. Molecular therapy with recombinant antisensec-myc adenovirus for human gastric carcinoma cells in vitro and in vivo[J]. JGastroenterol Hepatol,2001,16(1):22–28.
[41]Wong NA, Morris RG, McCondochie A, et al. Cyclin D1overexpression incolorectal carcinoma in vivo is dependent on beta-catenin protein dysregulation,but not k-ras mutation[J].J Pathol2002;197(1):128–135.
[42]Huang K, Zhang JX, Han L, et al. MicroRNA roles in beta-catenin pathway[J].Mol Cancer,2010,9:252.
[43] Hashimi ST, Fulcher JA, Chang MH, et al. MicroRNA profiling identifiesmiR-34a and miR-21and their target genes JAG1and WNT1in the coordinateregulation of dendritic cell differentiation[J]. Blood,2009,114(2):404–414.
[44]Burk U, Schubert J, Wellner U, et al. A reciprocal repression between ZEB1andmembers of the miR-200family promotes EMT and invasion in cancer cells[J].EMBO Rep,2008,9(6):582–589.
[45]Su J, Zhang A, Shi Z, et al. MicroRNA-200a suppresses the Wnt/β-cateninsignaling pathway by interacting with β-catenin[J]. Int J Oncol.2012,40(4):1162-1170.
[46]Xu J, Zhu X, Wu L, et al. MicroRNA-122suppresses cell proliferation andinduces cell apoptosis in hepatocellular carcinoma by directly targetingWnt/β-catenin pathway[J]. Liver Int,2012,32(5):752-760.
[47] Huang SD, Yuan Y, Zhuang CW, et al. MicroRNA-98and microRNA-214post-transcriptionally regulate enhancer of zeste homolog2and inhibit migrationand invasion in human esophageal squamous cell carcinoma[J]. Mol Cancer,2012,11:51.
[48]Zhao Z, Tan X, Zhao A, et al. microRNA-214-mediated UBC9expression inglioma[J]. BMB Rep,2012,45(11):641-646.
[49] Ueda T, Volinia S, Okumura H, et al. Relation between microRNA expressionand progression and prognosis of gastric cancer: a microRNA expressionanalysis[J]. Lancet Oncol,2010,11(2):136–146.
[50] Penna E, Orso F, Cimino D, et al. MicroRNA-214contributes to melanomatumour progression through suppression of TFAP2C[J]. EMBO J,2011,30(10):1990–2007.
[1] Chen Y, Shen A, Rider PJ, et al. A liver-specific microRNA binds to a highlyconserved RNA sequence of hepatitis B virus and negatively regulates viral geneexpression and replication [J]. FASEB J,2011,25(12):4511-4521.
[2] Zhang Y, Jia Y, Zheng R, et al. Plasma microRNA-122as a biomarker for viral-,alcohol-and chemical-related hepatic diseases [J]. Clin Chem,2010,56(12):1830-1838.
[3]贾音,张毅,费明钰,等.慢性乙肝患者外周血microRNAs的表达变化[J].第二军医大学学报,2010,31(12):1381-1383.
[4] Gui J, Tian Y, Wen X, et al. Serum microRNA characterization identifiesmiR-885-5p as a potential marker for detecting liver pathologies [J]. Clin Sci(Lond).,2011,120(5):183-193.
[5] Xu J, Wu C, Chen X, et al. Circulating microRNAs, miR-21, miR-122, andmiR-223, in patients with hepatocellular carcinoma or chronic hepatitis [J]. MolCarcinog,2011,50(2):136-142.
[6] Ji F, Yang B, Peng X, et al. Circulating microRNAs in hepatitis B virus-infectedpatients[J]. J Viral Hepat,2011,18(7): e242-251.
[7]张静,顾国浩,史进方,等.重型乙型肝炎患者外周血单个核细胞microRNA表达及其意义[J].临床检验杂志,2012,30(3):219-221.
[8] Wu FL, Jin WB, Li JH, et al. Targets for human encoded microRNAs in HBVgenes [J]. Virus Genes,2011,42(2):157-161.
[9] Zhang GL, Li YX, Zheng SQ, et al. Suppression of hepatitis B virus replicationby microRNA-199a-3p and microRNA-210[J]. Antiviral Res,2010,88(2):169-175.
[10]Zhang X, Zhang E, Ma Z, et al. Modulation of hepatitis B virus replication andhepatocyte differentiation by MircoRNA-1[J]. Hepatology,2011,53(5):1476-1485.
[11]郜玉峰,余莉,李家斌,等.靶向ASGPR1的外源性microRNA对HBV表达和复制的抑制作用[J].世界华人消化杂志,2009,17(7):699-704.
[12]李宁,陈明泉,程琦,等.调控慢性乙型肝炎患者I型干扰素信号通路的microRNA的筛选及鉴定[J].中国病毒病杂志,2012,2(2):107-112.
[13]Randall G, Panis M, Cooper JD, et al. Celluar cofactors affecting hepatitis C virusinfection and replication [J]. Proc Natl Acad Sci U S A,2007,104(31):12884-12889.
[14]Narbus CM, Israelow B, Sourisseau M, et al. HepG2cells expressing microRNAmiR-122support the entire hepatitis C virus life cycle [J]. J Virol,2011,85(22):12087-12092.
[15]Bihrer V, Friedrich-Rust M, Kronenberger B, et al. Serum miR-122as abiomarker of necroinflammation in patients with chronic hepatitis C virusinfection [J]. Am J Gastroenterol,2011,106(9):1663-1669.
[16]Sarason FM, Krol J, Markiewicz I, et al. Decreased levels of microRNA-122inindividuals with hepatitis C responding poorly to interferon therapy [J]. Nat Med,2009,15(1):31-33.
[17]Pedersen IM, Cheng G, Wieland S, et al. Interferon modulation of cellularmicroRNAs as an antiviral mechanism [J]. Nature,2007,449(7164):919-922.
[18]Scagnolari C, Zingariello P, Vecchiet J, et al. Differential expression ofinterferon-induced microRNAs in patients with chronic hepatitis C virus infectiontreated with pegylated interferon alpha [J]. Virol J,2010,(7):311.
[19]胡学玲. microRNA在乙型、丙型肝炎中的作用[J].临床肝胆病杂志,2012,28(11):877-880.
[20]Zhang Y, Jia Y, Zheng R, et al. Plasma microRNA-122as a biomarker for viral-,alcohol-, and chemical-related hepatic diseases [J]. Clin Chem,2010,56(12):1830-8.
[21]Wang K, Zhang S, Marzolf B, et al. Circulating microRNAs, potential biomarkersfor drug-induced liver injury [J]. Proc Natl Acad Sci USA,2009,106(11):4402-4407.
[22]Murakami Y, Toyoda H, Tanaka M, et al. The progression of liver fibrosis isrelated with overexpression of the miR-199and200families [J]. PLoS One,2011,6(1):e16081
[23]Roderburg C, Urban GW, Bettermann K, et al. Micro-RNA profiling reveals arole for miR-29in human and murine liver fibrosis [J]. Hepatology,2011,53(1):209-218.
[24]Li WQ, Chen C, Xu MD, et al. The rno-miR-34family is upregulated and targetsACSL1in dimethylnitrosamine-induced hepatic fibrosis in rats [J]. FEBS J.2011,278(9):1522-1532.
[25]Chen C, Wu CQ, Zhang ZQ, et al. Loss of expression of miR-335is implicated inhepatic stellate cell migration and activation. Exp Cell Res.2011,317(12):1714-1725.
[26]Furuta M, Ozawkie KI, Kanaka S, et al. miRNA-124and miRNA-203areepigenetically silenced tumor-suppressive Microfloras in hepatocellularcarcinoma [J]. Carcinogenesis,2010,31(5):766-776.
[27]Ivanovska I, Ball AS, Diaz RL, et al. MicroRNAs in the miR-106b familyregulate p21/CDKN1A and promote cell cycle progression[J]. Mol Cell Biol,2008,28(7):2167-2174.
[28]Fornent F, Lazzo M, Chico P, et al. miRNA-199a-3p regulates mTor and c-Met toinfluence the doxorubicin sensitivity of human adenocarcinomata cells[J]. CancerRes,2010,70(12):5184-5193.
[29]Hou J, Lin L, Zhou W, et al. Identification of miRNomes in human liver andhepatocellular carcinoma reveals miR-199a/b-3p as therapeutic target forhepatocellular carcinoma [J]. Cancer Cell,2011,19(2):232-243.
[30]Jia XQ, Cheng HQ, Qian X, et al. Lentivirus-mediated overexpression ofmicroRNA-199a inhibits cell proliferation of human hepatocellular carcinoma.[J].Cell Biochem Biophys,2012,62(1):237-244.
[31]Henry JC, Park JK, Jiang J, et al. MiR-199a-3p targets CD44and reducesproliferation of CD44positive hepatocellular carcinoma cell lines [J]. BiochemBiophys Res Commun,2010,403:120-125.
[32]Buurman R, Gurlevik E, Schaffer V, et al. Histone deacetylases activatehepatocyte growth factor signaling by repressing microRNA-449inhepatocellular carcinoma cells [J]. Gastroenterology,2012,[Epub ahead ofprint].
[33]Xu J, Zhu X, Wu L, et al. MicroRNA-122suppresses cell proliferation andinduces cell apoptosis in hepatocellular carcinoma by directly targetingWnt/β-catenin pathway [J]. Liver Int,2012,32(5):752-760.
[34]Chung GE, Mung SJ, Lee SH, et al. High expression of microRNA-15b predicts alow risk of tumor recurrence following curative resection of hepatocellularcarcinoma [J]. Colon Report,2010,23(1):113-119.
[35]Guo CJ, Pan Q, Li DG, et al. miR-15b and miR-16are implicated in activation ofthe rat hepatic stellate cell: An essential role for apoptosis [J]. J Hepatol,2009,50(4):766-778.
[36]张俊,罗娜,王勃,等.上调microRNA-150表达对肝癌SMMC7721细胞增殖和凋亡的影响[J].解放军医学杂志,2012,37(10):943-936.
[37]Li D, Liu X, Lin L, et al. MicroRNA-99a inhibits hepatocellular carcinomagrowth and correlates with prognosis of patients with hepatocellular carcinoma[J]. J Biol Chem,2011,286(42):36677-36685.
[38]张敬磊,金华君,刘辉,等.腺病毒介导microRNA-99a过表达抑制肝癌细胞生长[J].中国肿瘤生物治疗杂志,2012,19(3):267-271.
[39]Nohata N, Hanazawa T, Kikkawa N, et al. Tumor suppressive microRNA-375regulates oncogene AEG-1/MTDH in head and neck squamous cell carcinoma(HNSCC)[J]. J Human Genetics,2011,56(8):595-601.
[40]朱秀明. MicroRNAlet-7c在肝细胞癌中的表达、功能及靶点的研究[D].浙江大学,2012.
[41]Yao J, Liang L, Huang S, et al. MicroRNA-30d promotes tumor invasion andmetastasis by targeting Galphai2in hepatocellular carcinoma [J]. Hepatology,2010,51(3):846-856.
[42]Ding J, Huang S, Wu S, et al. Gain of miR-151on chromosome8q24.3facilitatestumour cell migration and spreading through downregulating RhoGDIA [J]. NatCell Biol,2010,12(4):390-399.
[43]Tsai WC, Lai TC, Chau GY, et al. MicroRNA-122，a tumor suppressor microRNAthat regulates intrahepatic metastasis of hepatocellular carcinoma [J]. Hepatology,2009,49(5):1571-1582.
[44]肖海静,王观宇,董庆华.人肝细胞肝癌和癌旁正常组织microRNA表达差异的分析[J].肿瘤防治研究,2012,39(8):947-949.
[45]Sukata T, Sumida K, Kushida M, et al. Circulating miRNAs, possible indicatorsof progress of rat hepatocarcinogenesis from early stages [J]. Toxicol Lett,2011,200(1-2):46-52.
[46]高军平,向邦德,倪航航,等. microRNA-224在肝细胞癌组织和血浆中表达的相关性[J].中国现代医学杂志,2011,32(21):4012-4021.
[47]Li LM, Hu ZB, Zhou ZX, et al. Serum miRNA profiles serve asnovel biomarkersfor HBV infection and diagnosis of HBV-positive hepatocarcinoma[J]. CancerRes,2010,70(23):9798－9807.
[48]Kota J, Chivukula RR, O'Donnell KA, et al. Therapeutic miRNA deliverysuppresses tumorigenesis in a murine liver cancer model [J]. Cell,2009,137(6):1005-1017.
[49]Connolly E, Melegari M, Landgraf P, et al.Elevated expression of the miR-17-92polycistron and miR-21in hepadnavirus-associated hepatocellular carcinomacontributes to the malignant phenotype[J]. Am J Pathol.2008,173(3):856-864.
[50]Zhou C, Liu J, Li Y, et al. microRNA-1274a, a modulator of sorafenib induced adisintegrin and metalloproteinase9(ADAM9) down-regulation in hepatocellularcarcinoma [J]. FEBS Lett,2011,585:1828-1834.
[51]Li J, Wang Y, Yu W, et al. Expression of yserum miRNA-221in humanhepatocellular carcinoma and its prognostic significance [J]. Biochem BiophysRes Commun,2011,406(1):70-73.
[52]Wang C, Song B, Song W, et al. Unpressed miRNA-199b-5p targetsHypoxia-lnducible Factor-1a in hepatocellular carcinoma and predicts prognosisof hepatocellular carcinoma patients [J]. J Gastroenteroi Hepatol,2011,26(11):1630-1637.
[53]Ji JF, Shi J, Budhu A, et al. miRNA expression, survival, and response tointerferon in liver cancer [J]. N Engl J Med,2009,361(15):1437-1447.
[54]倪航航,向邦德,高军平,等.血浆中microRNA-122与肝癌手术肝损伤的相关性研究[J].实用医学杂志,2012,28(6):932-935.
[55]邓治亮,孙建,区应亮,等.原发性肝癌术后早期复发密切相关microRNA的筛选及应用[J].中山大学学报(医学科学版),2012,33(4):494-498.
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