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目的观察健脾理气方药物血清对肝癌细胞 SMMC-7721端粒酶活性的影响以及诱导肿瘤细胞凋亡的作用,探讨其抗肿瘤治疗的作用机制。方法采用端粒重复序列扩增(TRAP)结合非变性聚丙烯酰胺凝胶电泳银染法,利用健脾理气方中药灌服新西兰兔后制备含药血清,观察健脾理气方药物血清对体外培养的人肝癌细胞株SMMC-7721端粒酶活性的影响,同时利用流式细胞仪检测和荧光显微镜观察对细胞凋亡的影响。结果健脾理气药物血清在 d4开始对肿瘤细胞的端粒酶活性有抑制作用,电泳条带光密度明显降低;并能诱导肿瘤细胞的凋亡,凋亡的比例随作用时间的延长而升高,流式细胞术检测d2,d4,d6的凋亡比例分别为0.81%,5.16%,8.45%。结论对肿瘤细胞端粒酶活性的抑制和诱导肿瘤细胞凋亡是键脾理气中药抗肿瘤治疗作用机制的一部分。
Objective To observe the effect of serum of Jianpi Liqi Prescription on the telomerase activity of hepatocellular carcinoma cell line SMMC-7721 and its role in inducing apoptosis of tumor cells, and to explore the mechanism of antitumor therapy. METHODS: Telomere-repeated sequence amplification (TRAP) combined with non-denaturing polyacrylamide gel electrophoresis silver-staining method was used to inoculate New Zealand rabbits with Chinese traditional medicine of Jianpi Liqi Decoction. The effects of telomerase activity on human hepatocellular carcinoma cell line SMMC-7721 were examined. Flow cytometry and fluorescence microscopy were used to observe the effect on cell apoptosis. Results Spleen and qi drugs serum inhibited the telomerase activity of tumor cells at the beginning of d4, the optical density of electrophoresis bands decreased significantly, and the apoptosis of tumor cells could be induced. The proportion of apoptosis increased with the extension of the action time. The percentages of apoptosis of d2, d4 and d6 detected by flow cytometry were 0.81%, 5.16% and 8.45%, respectively. Conclusion The inhibition of telomerase activity and the induction of tumor cell apoptosis in tumor cells are part of the mechanism of the antitumor effects of the key Spleen and Qi Chinese herbs.