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12-脂肪氧化酶抑制剂诱导人肝癌细胞凋亡及其对survivin基因表达的影响

赵继学1, 王广义2, 杨志明1   

  1. (1.吉林大学第一医院儿外科,吉林 长春 130021; 2.吉林大学第一医院普通外科,吉林 长春 130021)
  • 收稿日期:2007-03-29 修回日期:1900-01-01 出版日期:2007-11-28 发布日期:2007-11-28
  • 通讯作者: 杨志明

Apoptosis of human hepatocellular carcinoma cell induced by 12-lipoxygenase inhibitor and effect of 12-lipoxygenase inhibitor on expression of survivin gene

ZHAO Ji-xue1,WANG Guang-yi2,YANG Zhi-ming1   

  1. (1.Department of Pediatric Surgery,First Hospital,Jilin University,Changchun,130021,China ;2. Department of General Surgery,First Hospital,Jilin University,Changchun,130021,China)
  • Received:2007-03-29 Revised:1900-01-01 Online:2007-11-28 Published:2007-11-28
  • Contact: YANG Zhi-ming

摘要: 目的: 探讨12-脂肪氧化酶(12-LOX)抑制剂诱导人肝癌细胞凋亡的作用及其对survivin基因表达的影响。 方法: 对人肝癌细胞株HepG2进行细胞培养,取指数生长期细胞设对照组、12-LOX抑制剂组(baicalein 20、40和80 μmol•L-1组),加试剂后继续培养72 h。透射电镜观察抑制剂组和对照组细胞形态。RT-PCR法分别检测实验组和对照组肝癌细胞12-LOX mRNA的表达情况。MTT 法检测细胞增殖情况。DNA琼脂糖凝胶电泳检测细胞凋亡。RT-PCR法检测survivinmRNA的表达。结果: 人肝癌细胞株HepG2中存在12-LOX mRNA的明显表达,baicalein干预后,12-LOXmRNA表达量明显减弱。透射电镜下发现,对照组细胞胞核和细胞器亚微结构清晰,核膜完整;而经baicalein处理后,凋亡细胞增多,而且同一电镜切片中可以观察到不同时期的凋亡形态学改变,并可见坏死细胞及细胞碎片。MTT显示大致呈时间-剂量依赖性抑制细胞增殖,细胞存活率随着baicalein给药后时间的延长和给药浓度的增加而逐渐下降(P<0.05);baicalein 干预后HepG2细胞提取DNA进行琼脂糖凝胶电泳,可见梯形条带,而对照组未出现梯形条带。12-LOX抑制剂baicalein可降低HepG2细胞中survivin表达,并呈浓度-时间依赖性。结论: 12-LOX抑制剂能诱导人肝癌细胞HepG2的细胞凋亡,其诱导细胞凋亡的机制可能与下调抗凋亡基因survivin的表达有关。

关键词: 黄芩甙元, 肝肿瘤, 细胞凋亡, 逆转录聚合酶链反应

Abstract: To investigate the effect of 12-lipoxygenase(12-LOX) inhibitorinducing the apoptosis of human hepatocellular carcinoma cell line HepG2 on the expression of survivin gene. Methods HepG2 cells were cultivated in RPMI-1640 medium.12-LOX mRNA was analyzed by reverse transcription polymerase chain reaction (RT-PCR).The effect of baicalein on the proliferation of the cells was detected by thiazolyl blue tetrazoliumbromide (MTT) method.The subcellular structures were observed under electron microscope.DNA ladder pattern on agarose gel electrophores was used to evaluate the apoptotic index of hepG2 cells.The expression of survivin mRNA was analyzed by RT-PCR. Results12-LOX mRNA was expressed in human HepG2 cells.At concentrations from 20 to 80 μmol•L-1,baicalein inhibited the proliferation of HepG2 cells in a concentration- and time-dependent manner from 24 to 72 h.There were significant differences between any other groups (P<0.05).Morphological changes such as chromatin condensatio,apoptotic bodies were observed after treated with baicalein.Significant apoptosis was also induced by baicalein,and the apoptotic index was detected by agarose gel electrophores which showed eveident DNA fragmentation.12-LOX inhibitor depressed the expression of survivin mRNA in a concentration- and time-dependent manner from 24 to 72 h. Conclusion 12-LOX inhibitor can induce the the apoptosis of human hepatocellular carcinoma cell line HepG2,and the probable mechanism is to depress the expression of anti-apoptosis gene survivin.

Key words: baicalein, hepatic tumour, apoptosis, reverse transcription polymerase chain reaction

中图分类号: 

  • R735.7