摘要
目的从基因水平研究白花蛇舌草对体外培养的人肝癌Bel7402细胞诱导凋亡的作用及其可能的作用机制。方法采用人肝癌Bel7402细胞常规体外培养,随机设定空白对照组、5-氟尿嘧啶(5-Fu)阳性对照组、白花蛇舌草组。倒置显微镜直接观察细胞形态变化。HE染色法光镜下观察细胞形态改变,计算凋亡指数。采用MTT法检测白花蛇舌草对癌细胞的生长抑制作用。RT-PCR半定量法检测Bcl-xl、p53基因mRNA表达的变化。结果光镜下观察到白花蛇舌草组肿瘤细胞脱壁,有细胞出芽现象等凋亡形态改变;凋亡指数略低于5-Fu阳性对照组,但与空白对照组比较有显著差异(P<0.01);细胞抑制率略低于5-Fu阳性对照组,与空白对照组相比有显著差异(P<0.01);上调p53基因mRNA表达,降低Bcl-xl基因mRNA表达,与空白对照组比较均有显著差异。结论白花蛇舌草抑制人肝癌Bel7402细胞增长,诱导细胞凋亡,其分子机制可能与激活抑癌基因p53基因、抑制原癌基因Bcl-xl基因表达有关。
Objective To study the inducing apoptosis of Hedyotis diffusa extract （HDE） on human hepatocarcinoma cell line Bel7402 and its molecular mechanism. Methods Human hepatocarcinoma cell line Bel7402 was used in vitro cell culture, there were 3 groups： control group, HDE group and 5-Fu group, the Bel7402 cells were dealed with them respectively. The apoptosis morphologic changes of human hepatocarcinoma cells were observed by invert microscope. The apoptosis rate was detected with hematoxylin stain by light microscope. Inhibition of Bel7402 cell proliferation was measured by MTT assay. The expression of oncogene Bcl-x1 and anti-oncogene p53 of Bel7402 ceils were observed with RT-PCR assay. Results Compared with the control group, the condensation of nuclear, vacuolar degeneration of mitochondria could be found through light microscope. The apoptosis rate of HDE group was remarkably increased compared with that in control group （P〈0.01）. The inhibition of HDE group was remarkably increase compared with that in control group （P〈0.01）. The expression of the gene p53 raised and Bcl-x1 decreased. Conclusion HDP can induce tumor cells apoptosis, the activation of p53 and the suppression of Bcl-x1 may contribute to the apoptosis mechanism.
出处
《中国中医药信息杂志》
CAS
CSCD
2007年第11期33-35,共3页
Chinese Journal of Information on Traditional Chinese Medicine
基金
黑龙江省卫生厅课题(2600-400)
