小柴胡汤联合5-氟尿嘧啶诱导Hca-f肝癌小鼠细胞凋亡的实验研究
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摘要
目的:研究经典方剂小柴胡汤(以下简称xcht)联合化疗药物5-氟尿嘧啶(以下简称5-fu)对Hca-f肝癌小鼠的抑瘤作用及其作用机制。
材料与方法:取健康昆明种小鼠40只。在无菌条件下抽取Hca-f肝癌小鼠腹水为瘤源,用生理盐水稀释为4x10~6个瘤细胞/ml,每鼠给予0.2ml于其右侧腋窝皮下接种。24小时后随机分成单纯肿瘤组、xcht组、5-fu组、xcht+5-fu组,给药10天后,处死小鼠,剥离肿瘤,计算抑瘤率;HE染色观察肿瘤实质细胞变化;电镜观察肿瘤细胞凋亡:利用DNA原位末端标记法(TUNEL法)对肿瘤细胞凋亡计数;ELISA法检测血清中TNF-α浓度;TNF-α浓度与细胞凋亡的相关性分析。
结果:
1.各组小鼠一般状态:单纯肿瘤组小鼠毛色灰暗,运动迟缓;xcht组小鼠皮毛较光滑,运动略迟缓;5-fu组小鼠毛色稍暗,运动迟缓;xcht+5-fu组小鼠皮毛光滑,运动灵活。
2.经药物治疗后,Hca-f肝癌小鼠体内肿瘤生长受到不同程度的抑制,其抑瘤率分别为:xcht组31.88%,5-fu组37.50%,而xcht+5-fu组最高,可达50.80%。
3.光镜下可见单纯肿瘤组肿瘤组织排列紊乱,失去正常的结构与层次,肿瘤细胞大小不等,形状不规则,胞核深染,病理性核分裂像多见;xcht组、5-fu组肿瘤组织排列较规则,肿瘤细胞数目少于单纯肿瘤组;xcht+5-fu组肿瘤组织排列较规则,肿瘤细胞大小较一致,病理性核分裂像少见。
4.电镜下单纯肿瘤组可见到凋亡早期表现,肿瘤细胞膜失去连续性,核体积增大,异染色质增多成团块状,并出现边集现象;xcht组、5-fu组及xcht+5-fu组均可见到典型的凋亡小体,其中xcht+5-fu组数目最多。
5.TUNEL法检测凋亡细胞,各组细胞凋亡指数分别为:单纯肿瘤组2.10%,xcht组8.40%,5-fu组6.40%,而xcht+5-fu组最高,可达14.00%。
6.ELISA法测得各用药组的TNF-α浓度分别为:单纯肿瘤组99.91pg/ml,xcht组120.31pg/ml,5-fu组115.70pg/ml,而xcht+5-fu组最高,可达132.75pg/ml。
7.Hca-f肝癌小鼠血清中TNF-α浓度与细胞凋亡指数呈正相关。
结论:
1.小柴胡汤具有提高Hca-f肝癌小鼠生存质量的作用。
2.小柴胡汤具有抑制Hca-f肝癌小鼠肿瘤生长的作用。
3.小柴胡汤具有诱导肿瘤细胞凋亡的作用。
4.小柴胡汤具有促进Hca-f肝癌小鼠TNF-α产生的作用。
5.小柴胡汤提高Hca-f肝癌小鼠生存质量,抑制其肿瘤生长的作用是通过诱导肿瘤细胞凋亡,促进TNF-α产生的作用机制实现的。
6.Hca-f肝癌小鼠血清中TNF-α浓度与细胞凋亡指数呈正相关。
7.小柴胡汤与化疗药物5-fu联合使用,作用优于单独使用化疗药物。
Purpose: Study of classical Chinese medicine prescription of Xiaochaihu tang(later to be called xcht for short)combines with chemotherapy drug 5-fluorouracil (later to be called 5-fu) used in Hca-f liver cancer mouse damps the lump function and the function mechanism.
Material and method: Check 40 health female Kunming mice. For the mice collected in sterile conditions for the Hca-f liver cancer mouse ascites tumor source, diluted with NS for 4x10~6 tumor cells/ml, inoculated 0. 2ml per mouse to the right side of his armpit under the skin. After 24 hours were randomly divided into simple tumor group, xcht group, 5-fu group, xcht+5-fu group, 10 days after treatment, the mice were killed, stripped and weighing tumor weight to calculate the rate of tumor; by HE staining of tumor cells to see their changes; by electron microscope to see apoptosis; by the method of Tunnel to measured the rate of tumor cells' apoptosis; and use method ELISA to assay the serum of TNF-αin blood-serum; The serum of TNF-αand apoptosis in correlation analysis.
Results:
1. The general state of mice in each group: the simple tumor group have gloomy color fur and move slowly; xcht group have bright color fur, mental a little slow moves;5-fu group have a little gray color fur, and move slowly; xcht+5-fu group's mouse have bright and smooth fur, also more flexible move.
2. After drug treatment, Hca-f liver tumor growth in mice by different degrees of inhibition, the inhibition rates were: xcht group 31. 88%,5-fu group 37. 50%, while the xcht+5-fu group's inhibition rate is the highest of all, up to 50. 80%.
3. Through the light microscope, simple tumor group can be seen in tumor tissue with tumor group disorder, loss of normal structure and the level of tumor cell size, shape, irregular, deeply stained nuclei, also has pathological karyokinesis; xcht group and 5-fu group can be seen in tumor tissue with tumor group well ragulated, the number of tumor cells is less than the simple group; xcht+5-fu group, the tumor cells' organization with a certain regularity, more or less uniform, the size of tumor cells are more or less the same, pathological karyokinesis is much less than the simple group.
4. Under electron microscope, in the early time of apoptotic shows that, membranes lost continuty, nucleus volume become large, more heterochromatin become ball and in the side of nucleus. Typical apoptotic bodies can be seen in both xcht group, 5-fu group and xcht+5-fu group, the amount of apoptotic bodies which have been seen in xcht+5-fu group is more than other groups.
5. Use the method Tunnel to measured the fraction of tumor cells' apoptosis: The simple tumor group is 2. 10%, xcht group is 8. 40%, 5-fu group is 6. 40%,while the xcht+5-fu group is the highest, up to 14. 00%.
6. Use the method ELISA to measured the treatment group of TNF-αconcentrations: The simple tumor group is 99. 91pg/ml, xcht group is 120. 31pg/ml, 5-fu group is 115. 70pg/ml, while xcht+5-fu group is highest, up to 132. 75pg/ml.
7. Hca-f liver cancer in mice serum concentrations of TNF-αand apoptosis index were positively correlated.
Conclusion:
1. The prescription of Xiaochaihutang plays an important role in improving the Hca-f liver cancer mice's quality of life.
2. The prescription of Xiaochaihutang can inhibit tumor cell's growth.
3. The prescription of Xiaochaihutang has the function of promoting tumor cell apoptosis.
4. The prescription of Xiaochaihutang has promotes the function which the Hca-f liver cancer mouse TNF-αproduces.
5. The prescription of Xiaochaihutang can improve the Hca-f liver cancer mouse survival quality, suppresses the function which its tumor grows is through the induction tumour cell perishes weakly, the promotion TNF-αproduces the function mechanism realizes.
6. Hca-f liver cancer in mice serum concentrations of TNF-αand apoptosis index were positively correlated.
7. Use the prescription of Xiaochaihutang combined with chemotherapy drugs, has better effect than use chemotherapy drugs alone.
材料与方法:取健康昆明种小鼠40只。在无菌条件下抽取Hca-f肝癌小鼠腹水为瘤源,用生理盐水稀释为4x10~6个瘤细胞/ml,每鼠给予0.2ml于其右侧腋窝皮下接种。24小时后随机分成单纯肿瘤组、xcht组、5-fu组、xcht+5-fu组,给药10天后,处死小鼠,剥离肿瘤,计算抑瘤率;HE染色观察肿瘤实质细胞变化;电镜观察肿瘤细胞凋亡:利用DNA原位末端标记法(TUNEL法)对肿瘤细胞凋亡计数;ELISA法检测血清中TNF-α浓度;TNF-α浓度与细胞凋亡的相关性分析。
结果:
1.各组小鼠一般状态:单纯肿瘤组小鼠毛色灰暗,运动迟缓;xcht组小鼠皮毛较光滑,运动略迟缓;5-fu组小鼠毛色稍暗,运动迟缓;xcht+5-fu组小鼠皮毛光滑,运动灵活。
2.经药物治疗后,Hca-f肝癌小鼠体内肿瘤生长受到不同程度的抑制,其抑瘤率分别为:xcht组31.88%,5-fu组37.50%,而xcht+5-fu组最高,可达50.80%。
3.光镜下可见单纯肿瘤组肿瘤组织排列紊乱,失去正常的结构与层次,肿瘤细胞大小不等,形状不规则,胞核深染,病理性核分裂像多见;xcht组、5-fu组肿瘤组织排列较规则,肿瘤细胞数目少于单纯肿瘤组;xcht+5-fu组肿瘤组织排列较规则,肿瘤细胞大小较一致,病理性核分裂像少见。
4.电镜下单纯肿瘤组可见到凋亡早期表现,肿瘤细胞膜失去连续性,核体积增大,异染色质增多成团块状,并出现边集现象;xcht组、5-fu组及xcht+5-fu组均可见到典型的凋亡小体,其中xcht+5-fu组数目最多。
5.TUNEL法检测凋亡细胞,各组细胞凋亡指数分别为:单纯肿瘤组2.10%,xcht组8.40%,5-fu组6.40%,而xcht+5-fu组最高,可达14.00%。
6.ELISA法测得各用药组的TNF-α浓度分别为:单纯肿瘤组99.91pg/ml,xcht组120.31pg/ml,5-fu组115.70pg/ml,而xcht+5-fu组最高,可达132.75pg/ml。
7.Hca-f肝癌小鼠血清中TNF-α浓度与细胞凋亡指数呈正相关。
结论:
1.小柴胡汤具有提高Hca-f肝癌小鼠生存质量的作用。
2.小柴胡汤具有抑制Hca-f肝癌小鼠肿瘤生长的作用。
3.小柴胡汤具有诱导肿瘤细胞凋亡的作用。
4.小柴胡汤具有促进Hca-f肝癌小鼠TNF-α产生的作用。
5.小柴胡汤提高Hca-f肝癌小鼠生存质量,抑制其肿瘤生长的作用是通过诱导肿瘤细胞凋亡,促进TNF-α产生的作用机制实现的。
6.Hca-f肝癌小鼠血清中TNF-α浓度与细胞凋亡指数呈正相关。
7.小柴胡汤与化疗药物5-fu联合使用,作用优于单独使用化疗药物。
Purpose: Study of classical Chinese medicine prescription of Xiaochaihu tang(later to be called xcht for short)combines with chemotherapy drug 5-fluorouracil (later to be called 5-fu) used in Hca-f liver cancer mouse damps the lump function and the function mechanism.
Material and method: Check 40 health female Kunming mice. For the mice collected in sterile conditions for the Hca-f liver cancer mouse ascites tumor source, diluted with NS for 4x10~6 tumor cells/ml, inoculated 0. 2ml per mouse to the right side of his armpit under the skin. After 24 hours were randomly divided into simple tumor group, xcht group, 5-fu group, xcht+5-fu group, 10 days after treatment, the mice were killed, stripped and weighing tumor weight to calculate the rate of tumor; by HE staining of tumor cells to see their changes; by electron microscope to see apoptosis; by the method of Tunnel to measured the rate of tumor cells' apoptosis; and use method ELISA to assay the serum of TNF-αin blood-serum; The serum of TNF-αand apoptosis in correlation analysis.
Results:
1. The general state of mice in each group: the simple tumor group have gloomy color fur and move slowly; xcht group have bright color fur, mental a little slow moves;5-fu group have a little gray color fur, and move slowly; xcht+5-fu group's mouse have bright and smooth fur, also more flexible move.
2. After drug treatment, Hca-f liver tumor growth in mice by different degrees of inhibition, the inhibition rates were: xcht group 31. 88%,5-fu group 37. 50%, while the xcht+5-fu group's inhibition rate is the highest of all, up to 50. 80%.
3. Through the light microscope, simple tumor group can be seen in tumor tissue with tumor group disorder, loss of normal structure and the level of tumor cell size, shape, irregular, deeply stained nuclei, also has pathological karyokinesis; xcht group and 5-fu group can be seen in tumor tissue with tumor group well ragulated, the number of tumor cells is less than the simple group; xcht+5-fu group, the tumor cells' organization with a certain regularity, more or less uniform, the size of tumor cells are more or less the same, pathological karyokinesis is much less than the simple group.
4. Under electron microscope, in the early time of apoptotic shows that, membranes lost continuty, nucleus volume become large, more heterochromatin become ball and in the side of nucleus. Typical apoptotic bodies can be seen in both xcht group, 5-fu group and xcht+5-fu group, the amount of apoptotic bodies which have been seen in xcht+5-fu group is more than other groups.
5. Use the method Tunnel to measured the fraction of tumor cells' apoptosis: The simple tumor group is 2. 10%, xcht group is 8. 40%, 5-fu group is 6. 40%,while the xcht+5-fu group is the highest, up to 14. 00%.
6. Use the method ELISA to measured the treatment group of TNF-αconcentrations: The simple tumor group is 99. 91pg/ml, xcht group is 120. 31pg/ml, 5-fu group is 115. 70pg/ml, while xcht+5-fu group is highest, up to 132. 75pg/ml.
7. Hca-f liver cancer in mice serum concentrations of TNF-αand apoptosis index were positively correlated.
Conclusion:
1. The prescription of Xiaochaihutang plays an important role in improving the Hca-f liver cancer mice's quality of life.
2. The prescription of Xiaochaihutang can inhibit tumor cell's growth.
3. The prescription of Xiaochaihutang has the function of promoting tumor cell apoptosis.
4. The prescription of Xiaochaihutang has promotes the function which the Hca-f liver cancer mouse TNF-αproduces.
5. The prescription of Xiaochaihutang can improve the Hca-f liver cancer mouse survival quality, suppresses the function which its tumor grows is through the induction tumour cell perishes weakly, the promotion TNF-αproduces the function mechanism realizes.
6. Hca-f liver cancer in mice serum concentrations of TNF-αand apoptosis index were positively correlated.
7. Use the prescription of Xiaochaihutang combined with chemotherapy drugs, has better effect than use chemotherapy drugs alone.
引文
[1]孙震晓,赵天德,魏育林,等.去甲斑蝥素诱导人肝癌BEL-7402细胞凋亡的研究[J].解剖学报.1999,30(1):65-68
[2]Yan F,Tian XM,Ma XD.Effects of resveratrol on growth inhibition and gap-junctional intercellulax communication of HepG2 cells[J].Nanfang Yike Daxue Xuebao 2006,26:963-966
[3]赵建斌,崔勤,张雪,等.华蟾毒精抗癌作用的体外研究[J].第四军医大学学报.2001,22(16):1504-1506
[4]杨玉秀,徐秋霞,李修玲,等.猪苓多糖脂质体激活肝癌患者腹腔巨噬细胞抗瘤作用[J].中华肝脏病杂志.1998,6(4):220
[5]Shen YM,Zhu X,Zhang KH,Xie Y,Chen J,Dai Y,Ou-Yang CH,Li BM.Effect of ursolic acid on proliferation and apoptosis of hepatic stellate cells in vitro[J].Zhonghua Ganzangbing Zazhi.2008,16:298-301
[6]丁志山,高承贤,陈铌铍,等.姜黄素具有抑制血管生成与诱导肿瘤细胞凋亡双重作用[J].中国药理学通报.2003,19(2):171-173
[7]韩克起,黄传继.扶正抗癌方对大鼠移植性肝癌病理形态及超微结构的影响[J].中西医结合肝病杂志.1999,9(5):25-26
[8]陈林香,戴馨仪,周岱翰,等.清金得生片对小鼠移植性肝癌细胞增生与凋亡的影响[J].肿瘤研究与临床.2003,15(3):153-156
[9]张天娥,骆永珍,闫智勇.鹤莲克癌灵对肝癌H22细胞周期及其细胞凋亡的影响[J].成都中医药大学学报.1999,22(4):22-23
[10]李东涛,孙桂芝,裴迎霞,等.益气活血软坚解毒方含药血清诱导人肝癌细胞系Bel-7402细胞的凋亡[J].世界华人消化杂志.2005,13:2217-2221
[11]刘华,毛小玲,钟海波.晚癌康2号对原发性肝癌细胞凋亡基因bcl-2,Bax和fas表达的影响[J].中医药学刊.2005,23:1409-1410
[12]董青.百福生胶囊诱导小鼠肝癌细胞凋亡及对相关基因表达影响的实验研究[J].河南中医学院学报.2006,21:84
[13]赵晓琴.复方龙葵糖浆诱导小鼠H22肝癌细胞凋亡作用机制的研究[J].辽宁中医杂志.2005,32:974-975
[14]林云峰,明月,李靖涛.西洋参多糖Ⅰ、Ⅱ、Ⅲ对肝癌细胞DNA合成抑制作用的研究[J].临床肝胆杂志.1999,15(4):213-214
[15]司维柯,李鹏,姚婕.苦参碱对HepG2细胞代谢水平和基因水平的影响[J].第三军医大学学报.2002,24(11):1346-1349
[16]罗民,盛辉,王医林.红景天对肝癌细胞增殖的抑制作用[J].吉林医学.2005,26:1285-1286
[17]丁晓明,成肇仁.麝黄消瘤方抑制肝癌细胞侵袭粘附作用的实验研究[J].中西医结合肝病杂志.2006,16(2):96-98
[18]张晨,吕书琴,张亚妮,等.几种中药注射剂体外对肝癌细胞Bel-7042的杀伤作用[J].浙江中医学院学报.2005,29(6):93-95
[19]解方为,欧阳学农,蒋明德.红景天甙对人肝癌细胞的诱导分化作用[J].西南国防医药.2005,15:613-615
[20]曾小莉。涂植光.端粒酶在人参皂甙Rh-2诱导肝癌细胞分化中的作用[J].癌症.2005,23: 879-884
[21]蓝惠玲,王昌俊,刘友章,等.健脾化瘀中药逆转人肝癌细胞恶性表型作用研究[J].中医药通报.2006,12:5651
[22]刘平,周建峰,胡义扬,等.益气益阴诱导SMMC-7721肝癌细胞分化作用与意义[J].中国中医基础医学杂志.2000,6(8):9-34
[23]曾建新,王文亮,王知力,等.砷诱导的肝癌细胞凋亡及端粒酶活性改变[J].卫生毒理学杂志.2002。16(2):88-90
[24]孟志强,于尔辛,宋明志,等.健脾理气中药对肝癌端粒酶活性的影响[J].中国中医基础医学杂志.2000,6(1):23-26
[25]蒋小玲,邓欣,聂广,等.剔毒护肝方及其拆方对人肝癌细胞增殖及端粒酶活性的影响[J].中西医结合肝病杂志.2004,14(1):92-95
[26]邓欣,杨大国,吴其恺,等.正肝方及其拆方影响人肝癌细胞增殖及端粒酶活性的体外实验研究[J].中国中西医结合急救杂志.2005,12:338-340
[27]Singh F,Gao D,Lebwohl MG,Wei H.Shikonin modulates cell proliferation by inhibiting epidermal growth factor receptor signaling in human epidermoid carcinoma cells[J].Cancer Lett.2003,200:l15-121
[28]殷飞,吴新满,高洪生,等.清肝化瘀方含药血清对TGF-α诱导SMMC-7721细胞Raf-MEK-ERK信号传导的影响[J].世界华人消化杂志.2005,13:88-90
[29]许杜娟,陈敏珠.黄芪多糖的抑瘤作用及其机制[J].中国医院药学杂志.2005,25:923-925
[30]陈涛,胡卫,崔邦平,等.珠子参对小鼠H22肝癌抑制作用及机制[J].世界华人消化杂志.2007,15:2597-2601
[31]Yang JZ,Zhuo J,Chen BK,Jin LQ,Lv JX,LiLJ.Regulating effects of Paecilomyces cicadae polysaccharides on immunity of aged rats[J].Zhongguo Zhongyao Zazhi 2008,33:292-295
[32]王大宪,吴晓丰,郑沁荣,等.化徵丸抗小鼠H22肝癌的效应及对淋巴细胞转化率的影响[J].中西医结合肝病杂志.2005,15:97-100
[33]刘明华,章卓,顾立,等.伍氏百叶散对荷瘤小鼠肿瘤生长及免疫功能的影响[J].中国新药杂志.2006,15:201-204
[34]曹文涛,石巍.白藜芦醇抗肿瘤血管生成机制[J].国际肿瘤学杂志.2006,33:42-45
[35]Yi T,Yi Z,Cho SG,Luo J,Pandey MI,Aggarwal BB,Liu M.Gambogic acid inhibits angiogenesis and prostate tumor growth by suppressing vascular endothelial growth factor receptor 2 signaling[J].Cancer Res.2008,68:1843-1850
[36]殷飞,姚树坤,吴新满.清肝化瘀方药、苦参素对大鼠肝癌形成过程中ERK蛋白的影响[J].中国中医基础医学杂志.2006,12:270-275
[37]王庆高,韦艾凌,徐志新.癌痛消胶囊调节小鼠H-22移植性肝癌细胞荷瘤VEGF、p53、p2lras表达的实验研究[J].广西中医药.2005,27:45-48
[38]夏荣龙,陈存平.丹参对肝癌致鼠梗阻性黄疸时VEGF表达的影响[J].第四军医大学学报.2005.26:1673-1675
[39]候华新,黎丹戎,韦长元,等.板兰根高级不饱和脂肪酸对耐药肝癌细胞株BEL-7404逆转作用实验[J].中国现代应用药学.2002,19(5):351-353
[40]魏志霞.川芎嗪对肝癌多药耐药株SMMC-7721/ADM的逆转作用[J].江苏医药.2005,31:371-372
[41]李起,刘作金,张俊,等.中药复方肝癌1号逆转肝癌多药耐药的实验研究[J].消化外科.2006,5:70-73
[42]林丽珠.中医药在原发性肝癌综合治疗中对生存质量维护作用[J].中华肿瘤杂志.2003,25(2):202-203
[43]钱新华,左寿海.健脾增免冲剂的抗癌作用及其对免疫功能的影响[J].中药材.1998,21(7):359-360
[44]许得盛,沈自尹,王文健,等.右归饮、四君子汤、桃红四物汤调节肾虚、脾虚、血瘀证患者免疫功能的观察[J].中国中西医结合杂志.1999,19(12):712-714
[45]黄智芬,黎汉忠,施智严,等.健脾消积口服液结合西药治疗Ⅱ、Ⅲ期肝癌40例的临床观察[J].中国中西医结合杂志.2003,23(3):192-194
[46]纪均,曹杰,夏宝泉.肝动脉介入法联合“金米颗粒冲剂”治疗肝癌及其对细胞免疫功能的影响[J].临床肿瘤学杂志.2001,6(3):265-266.
[47]付于,夏天.消瘤汤治疗肿瘤及调节免疫功能的实验研究[J].中国中西医结合杂志.2004,24(12):1114-1115.
[48]陈长怀,宋莉,杨国风.青龙丸治疗原发性肝癌疗教观察[J].中医杂志.1996,37(3):165
[49]黄立中,田辉原发性肝癌中医治法的临床研究[J].湖南中医学院学报.1996,16(3):14
[50]朱洪恩,宋慧敏,单国英.华虎内功汤及热敷稍瘤散治疗原发性肝癌118例[J].江苏中医.1996,17(7):22
[51]孙开玲,彭立文,李爱玲.膈下逐瘀汤治疗原发性肝癌40例[J].中国民间疗法.1998,28(2):34
[52]王献民,刘敛,周心芳.八珍神农饮治疗原发性肝癌临床观察[J].河南中医药学 刊.1996,11(4):34
[53]王榕平,王莹.消肝瘤合剂治疗原发性肝癌近期疗教观察[J].福建中医药.1997,28(3):1
[54]陈群雄.参三金粉对肝癌介入化疗后的作用[J].中医药研究.1998,14(3):34
[55]吕瑞民.养肝抗癌丸配合肝动脉化疗栓塞治疗原发性肝癌32例临床分析[J].中医药信息.1997,6(2):26
[56]邵中兴,程志刚,尹学恩,等.肝动脉化疗栓塞术联合中药肝癌Ⅰ号、Ⅱ号治疗中晚期肝癌的临床观察[J].中国中西医结合杂志.2001,2l(3):168-170
[57]冯坚.中药和介入治疗原发性肝癌35例[J].广西中医学院学报.2002,5(3):51-52
[58]吴洁清,姚卫华,杨瑞明,等.安络化纤丸联合TACE术治疗原发性肝癌临床观察[J].中国中医药信息杂志.2005,12(8):559-563
[59]韩铭钧,任克,赵仲春,等.油性中药抗癌剂OCC肝动脉栓塞治疗肝癌84例3年随访报告[J].临床放射学杂志.1998,17(2):112-114
[60]范忠泽,沈建华,孙钰,等.中药消癌平针剂经肝动脉介入治疗晚期肝癌的临床观察[J].上海医药.1997,5:15-19
[61]王安浦,李捷.大剂量清开灵注射液在肝动脉栓塞中的应用[J].上海中医药杂志.1997,(6):封三
[62]盛淑芬.中西药台用治疗晚期肝癌26侧观察[J].实用中医药杂志.1998,14(8):7
[63]邵世祥,袁海燕,闰惠君,等.连慈饮治疗原发性肝癌临床观察[J].实用中医内科杂志.2005,19(2):157
[64]陈洪国,杜晓军.内外合治中晚期肝癌30例[J].山西中医.2005,21(5):15-16
[65]林志智,陈玲玲.小柴胡汤加味合消症益肝片联合化学药物治疗原发性肝癌13例[J].福建中医药.1996,27(1):8
[66]李永安,魏子祥,王里,等.三维适形放疗联合中药治疗肝癌的疗效观察[J].北京中医药大学学报.2004,11(3):18-20
[67]李增灿,陈志彪,孟庆峰,等.序贯治疗中、晚期肝癌的机理探讨[J].医师进修杂志.1999,22(8):28-29
[68]张成阳.中西医结合治疗晚期肝癌疗效观察[J].现代肿瘤医学,2003,11(5):380-381
[69]陈黠,翟笑枫,蒋栋,等.去甲斑螯素一泊洛沙姆407缓释剂和无水乙醇的肝癌瘤内注射疗效比较[J].第二军医大学学报.2001,22(7):606-608
[70]嵇玉峰.大承气汤滴肛治疗原发性肝癌并发肝性脑病38例临床疗教观察[J].中医药研究.1999,14(4):29
[71]刘毅斌.中药灌肠治疗原发性肝癌并肝性脑病63例[J].广西中医药.1999,20(2):16
[72]金红.泻心汤为主治疗中晚期肝癌上消化道出血40例临床观察[J].中国中西医结合杂志.1996,16(12):743
[73]朱玉明.玉枢丹外救缓解肝癌疼痛临床观察[J].中医外科杂志.1996,5(1):42
[74]王锦鸿.中医在治疗恶性肿瘤中的作用与地位[J].江苏中医.2001,22(11):1-3
[2]Yan F,Tian XM,Ma XD.Effects of resveratrol on growth inhibition and gap-junctional intercellulax communication of HepG2 cells[J].Nanfang Yike Daxue Xuebao 2006,26:963-966
[3]赵建斌,崔勤,张雪,等.华蟾毒精抗癌作用的体外研究[J].第四军医大学学报.2001,22(16):1504-1506
[4]杨玉秀,徐秋霞,李修玲,等.猪苓多糖脂质体激活肝癌患者腹腔巨噬细胞抗瘤作用[J].中华肝脏病杂志.1998,6(4):220
[5]Shen YM,Zhu X,Zhang KH,Xie Y,Chen J,Dai Y,Ou-Yang CH,Li BM.Effect of ursolic acid on proliferation and apoptosis of hepatic stellate cells in vitro[J].Zhonghua Ganzangbing Zazhi.2008,16:298-301
[6]丁志山,高承贤,陈铌铍,等.姜黄素具有抑制血管生成与诱导肿瘤细胞凋亡双重作用[J].中国药理学通报.2003,19(2):171-173
[7]韩克起,黄传继.扶正抗癌方对大鼠移植性肝癌病理形态及超微结构的影响[J].中西医结合肝病杂志.1999,9(5):25-26
[8]陈林香,戴馨仪,周岱翰,等.清金得生片对小鼠移植性肝癌细胞增生与凋亡的影响[J].肿瘤研究与临床.2003,15(3):153-156
[9]张天娥,骆永珍,闫智勇.鹤莲克癌灵对肝癌H22细胞周期及其细胞凋亡的影响[J].成都中医药大学学报.1999,22(4):22-23
[10]李东涛,孙桂芝,裴迎霞,等.益气活血软坚解毒方含药血清诱导人肝癌细胞系Bel-7402细胞的凋亡[J].世界华人消化杂志.2005,13:2217-2221
[11]刘华,毛小玲,钟海波.晚癌康2号对原发性肝癌细胞凋亡基因bcl-2,Bax和fas表达的影响[J].中医药学刊.2005,23:1409-1410
[12]董青.百福生胶囊诱导小鼠肝癌细胞凋亡及对相关基因表达影响的实验研究[J].河南中医学院学报.2006,21:84
[13]赵晓琴.复方龙葵糖浆诱导小鼠H22肝癌细胞凋亡作用机制的研究[J].辽宁中医杂志.2005,32:974-975
[14]林云峰,明月,李靖涛.西洋参多糖Ⅰ、Ⅱ、Ⅲ对肝癌细胞DNA合成抑制作用的研究[J].临床肝胆杂志.1999,15(4):213-214
[15]司维柯,李鹏,姚婕.苦参碱对HepG2细胞代谢水平和基因水平的影响[J].第三军医大学学报.2002,24(11):1346-1349
[16]罗民,盛辉,王医林.红景天对肝癌细胞增殖的抑制作用[J].吉林医学.2005,26:1285-1286
[17]丁晓明,成肇仁.麝黄消瘤方抑制肝癌细胞侵袭粘附作用的实验研究[J].中西医结合肝病杂志.2006,16(2):96-98
[18]张晨,吕书琴,张亚妮,等.几种中药注射剂体外对肝癌细胞Bel-7042的杀伤作用[J].浙江中医学院学报.2005,29(6):93-95
[19]解方为,欧阳学农,蒋明德.红景天甙对人肝癌细胞的诱导分化作用[J].西南国防医药.2005,15:613-615
[20]曾小莉。涂植光.端粒酶在人参皂甙Rh-2诱导肝癌细胞分化中的作用[J].癌症.2005,23: 879-884
[21]蓝惠玲,王昌俊,刘友章,等.健脾化瘀中药逆转人肝癌细胞恶性表型作用研究[J].中医药通报.2006,12:5651
[22]刘平,周建峰,胡义扬,等.益气益阴诱导SMMC-7721肝癌细胞分化作用与意义[J].中国中医基础医学杂志.2000,6(8):9-34
[23]曾建新,王文亮,王知力,等.砷诱导的肝癌细胞凋亡及端粒酶活性改变[J].卫生毒理学杂志.2002。16(2):88-90
[24]孟志强,于尔辛,宋明志,等.健脾理气中药对肝癌端粒酶活性的影响[J].中国中医基础医学杂志.2000,6(1):23-26
[25]蒋小玲,邓欣,聂广,等.剔毒护肝方及其拆方对人肝癌细胞增殖及端粒酶活性的影响[J].中西医结合肝病杂志.2004,14(1):92-95
[26]邓欣,杨大国,吴其恺,等.正肝方及其拆方影响人肝癌细胞增殖及端粒酶活性的体外实验研究[J].中国中西医结合急救杂志.2005,12:338-340
[27]Singh F,Gao D,Lebwohl MG,Wei H.Shikonin modulates cell proliferation by inhibiting epidermal growth factor receptor signaling in human epidermoid carcinoma cells[J].Cancer Lett.2003,200:l15-121
[28]殷飞,吴新满,高洪生,等.清肝化瘀方含药血清对TGF-α诱导SMMC-7721细胞Raf-MEK-ERK信号传导的影响[J].世界华人消化杂志.2005,13:88-90
[29]许杜娟,陈敏珠.黄芪多糖的抑瘤作用及其机制[J].中国医院药学杂志.2005,25:923-925
[30]陈涛,胡卫,崔邦平,等.珠子参对小鼠H22肝癌抑制作用及机制[J].世界华人消化杂志.2007,15:2597-2601
[31]Yang JZ,Zhuo J,Chen BK,Jin LQ,Lv JX,LiLJ.Regulating effects of Paecilomyces cicadae polysaccharides on immunity of aged rats[J].Zhongguo Zhongyao Zazhi 2008,33:292-295
[32]王大宪,吴晓丰,郑沁荣,等.化徵丸抗小鼠H22肝癌的效应及对淋巴细胞转化率的影响[J].中西医结合肝病杂志.2005,15:97-100
[33]刘明华,章卓,顾立,等.伍氏百叶散对荷瘤小鼠肿瘤生长及免疫功能的影响[J].中国新药杂志.2006,15:201-204
[34]曹文涛,石巍.白藜芦醇抗肿瘤血管生成机制[J].国际肿瘤学杂志.2006,33:42-45
[35]Yi T,Yi Z,Cho SG,Luo J,Pandey MI,Aggarwal BB,Liu M.Gambogic acid inhibits angiogenesis and prostate tumor growth by suppressing vascular endothelial growth factor receptor 2 signaling[J].Cancer Res.2008,68:1843-1850
[36]殷飞,姚树坤,吴新满.清肝化瘀方药、苦参素对大鼠肝癌形成过程中ERK蛋白的影响[J].中国中医基础医学杂志.2006,12:270-275
[37]王庆高,韦艾凌,徐志新.癌痛消胶囊调节小鼠H-22移植性肝癌细胞荷瘤VEGF、p53、p2lras表达的实验研究[J].广西中医药.2005,27:45-48
[38]夏荣龙,陈存平.丹参对肝癌致鼠梗阻性黄疸时VEGF表达的影响[J].第四军医大学学报.2005.26:1673-1675
[39]候华新,黎丹戎,韦长元,等.板兰根高级不饱和脂肪酸对耐药肝癌细胞株BEL-7404逆转作用实验[J].中国现代应用药学.2002,19(5):351-353
[40]魏志霞.川芎嗪对肝癌多药耐药株SMMC-7721/ADM的逆转作用[J].江苏医药.2005,31:371-372
[41]李起,刘作金,张俊,等.中药复方肝癌1号逆转肝癌多药耐药的实验研究[J].消化外科.2006,5:70-73
[42]林丽珠.中医药在原发性肝癌综合治疗中对生存质量维护作用[J].中华肿瘤杂志.2003,25(2):202-203
[43]钱新华,左寿海.健脾增免冲剂的抗癌作用及其对免疫功能的影响[J].中药材.1998,21(7):359-360
[44]许得盛,沈自尹,王文健,等.右归饮、四君子汤、桃红四物汤调节肾虚、脾虚、血瘀证患者免疫功能的观察[J].中国中西医结合杂志.1999,19(12):712-714
[45]黄智芬,黎汉忠,施智严,等.健脾消积口服液结合西药治疗Ⅱ、Ⅲ期肝癌40例的临床观察[J].中国中西医结合杂志.2003,23(3):192-194
[46]纪均,曹杰,夏宝泉.肝动脉介入法联合“金米颗粒冲剂”治疗肝癌及其对细胞免疫功能的影响[J].临床肿瘤学杂志.2001,6(3):265-266.
[47]付于,夏天.消瘤汤治疗肿瘤及调节免疫功能的实验研究[J].中国中西医结合杂志.2004,24(12):1114-1115.
[48]陈长怀,宋莉,杨国风.青龙丸治疗原发性肝癌疗教观察[J].中医杂志.1996,37(3):165
[49]黄立中,田辉原发性肝癌中医治法的临床研究[J].湖南中医学院学报.1996,16(3):14
[50]朱洪恩,宋慧敏,单国英.华虎内功汤及热敷稍瘤散治疗原发性肝癌118例[J].江苏中医.1996,17(7):22
[51]孙开玲,彭立文,李爱玲.膈下逐瘀汤治疗原发性肝癌40例[J].中国民间疗法.1998,28(2):34
[52]王献民,刘敛,周心芳.八珍神农饮治疗原发性肝癌临床观察[J].河南中医药学 刊.1996,11(4):34
[53]王榕平,王莹.消肝瘤合剂治疗原发性肝癌近期疗教观察[J].福建中医药.1997,28(3):1
[54]陈群雄.参三金粉对肝癌介入化疗后的作用[J].中医药研究.1998,14(3):34
[55]吕瑞民.养肝抗癌丸配合肝动脉化疗栓塞治疗原发性肝癌32例临床分析[J].中医药信息.1997,6(2):26
[56]邵中兴,程志刚,尹学恩,等.肝动脉化疗栓塞术联合中药肝癌Ⅰ号、Ⅱ号治疗中晚期肝癌的临床观察[J].中国中西医结合杂志.2001,2l(3):168-170
[57]冯坚.中药和介入治疗原发性肝癌35例[J].广西中医学院学报.2002,5(3):51-52
[58]吴洁清,姚卫华,杨瑞明,等.安络化纤丸联合TACE术治疗原发性肝癌临床观察[J].中国中医药信息杂志.2005,12(8):559-563
[59]韩铭钧,任克,赵仲春,等.油性中药抗癌剂OCC肝动脉栓塞治疗肝癌84例3年随访报告[J].临床放射学杂志.1998,17(2):112-114
[60]范忠泽,沈建华,孙钰,等.中药消癌平针剂经肝动脉介入治疗晚期肝癌的临床观察[J].上海医药.1997,5:15-19
[61]王安浦,李捷.大剂量清开灵注射液在肝动脉栓塞中的应用[J].上海中医药杂志.1997,(6):封三
[62]盛淑芬.中西药台用治疗晚期肝癌26侧观察[J].实用中医药杂志.1998,14(8):7
[63]邵世祥,袁海燕,闰惠君,等.连慈饮治疗原发性肝癌临床观察[J].实用中医内科杂志.2005,19(2):157
[64]陈洪国,杜晓军.内外合治中晚期肝癌30例[J].山西中医.2005,21(5):15-16
[65]林志智,陈玲玲.小柴胡汤加味合消症益肝片联合化学药物治疗原发性肝癌13例[J].福建中医药.1996,27(1):8
[66]李永安,魏子祥,王里,等.三维适形放疗联合中药治疗肝癌的疗效观察[J].北京中医药大学学报.2004,11(3):18-20
[67]李增灿,陈志彪,孟庆峰,等.序贯治疗中、晚期肝癌的机理探讨[J].医师进修杂志.1999,22(8):28-29
[68]张成阳.中西医结合治疗晚期肝癌疗效观察[J].现代肿瘤医学,2003,11(5):380-381
[69]陈黠,翟笑枫,蒋栋,等.去甲斑螯素一泊洛沙姆407缓释剂和无水乙醇的肝癌瘤内注射疗效比较[J].第二军医大学学报.2001,22(7):606-608
[70]嵇玉峰.大承气汤滴肛治疗原发性肝癌并发肝性脑病38例临床疗教观察[J].中医药研究.1999,14(4):29
[71]刘毅斌.中药灌肠治疗原发性肝癌并肝性脑病63例[J].广西中医药.1999,20(2):16
[72]金红.泻心汤为主治疗中晚期肝癌上消化道出血40例临床观察[J].中国中西医结合杂志.1996,16(12):743
[73]朱玉明.玉枢丹外救缓解肝癌疼痛临床观察[J].中医外科杂志.1996,5(1):42
[74]王锦鸿.中医在治疗恶性肿瘤中的作用与地位[J].江苏中医.2001,22(11):1-3