化疗药物对人肝癌BEL-7404细胞端粒的作用

Effects of chemotherapeutic drugs on telomer of human liver cancer BEL-7404 cell

目的观察几种常用化疗药物对人肝癌BEL-7404细胞端粒的作用。方法利用MTT法、端粒重复扩增实验(TRAP)法、RT-PCR、SouthernBlot和荧光染色法,分别测定经过不同浓度和作用时间多种化疗药物(顺铂、表柔比星、丝裂霉素、长春新碱)作用后BEL-7404端粒的变化。结果化疗药浓度越高和作用时间越长,端粒酶活性抑制、hTERT表达下调和端粒平均长度缩短越显著,其中顺铂和丝裂霉素作用最强。高浓度顺铂在48h(活性=0.18±0.14,t=85.42,P=0.000)、丝裂霉素在72h(活性=0.15±0.02,t=15.2,P=0.004)后端粒酶活性完全抑制,其余各组均未达完全抑制。中低浓度长春新碱对端粒长度无明显抑制(t=3.883,P=0.06;t=4.171,P=0.053),其余各组均有显著抑制。结论化疗药对BEL-7404细胞端粒有抑制作用,抑制作用强弱与药物种类、浓度和作用时间有关。

OBJECTIVE：To investigate the effects of several chemotherapeutic drugs on telomer of human liver cancer BEL-7404 cells. METHODS： By MTT assay, telomerase repeat amplification protocol （TRAP）, RT-PCR, Southern Blot and immuno-fluorescence, the changes of telomer of human liver cancer cell line BEL-7404 were detected, after being given cisplatin, vincristine, doxorubicin, mitomycin C at different concentrations and over different periods. RESULTS： The higher concentration and longer period of drugs were given, the stronger inhibition of telomerase activity, down regulation of bTERT and shortening of telomer would appeared. Cisplatin and mitomycin C showed the strongest activity. Cisplatin and mitomycin C of the high concentration group inhibited activity of telomere completely after 48 and 72 hours separately （activity=0.18± 0.14,t=85,42,P=0.000; activity = 0.15 ± 0. 02 , t = 15.2, P= 0. 004）, while no complete inhibition was seen in other groups. Vincristine of moderate and low concentration group showed no significant effect on shortening telomer （t 3.883, P=0.06; t=4.171, P= 0. 053）, while the rest groups revealed obvious effects. CONCLUSION： Chemotherapeutic drugs can suppress telomer of human liver cancer BEL-7404 cell, which correlates with category, concentration and period of drug.