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目的研究雷帕霉素联合顺铂诱导肝癌细胞凋亡、自噬及其对细胞侵袭转移能力的影响。方法将雷帕霉素以及顺铂分别或联合作用于HepG-2细胞,采用MTT法检测HepG-2细胞增殖抑制情况,采用流式细胞仪检测HepG-2细胞凋亡,采用MDC染色检测细胞自噬,用Transwell小室测定HepG-2细胞对细胞外基质的侵袭力。结果与对照组相比,单独应用雷帕霉素组未出现明显的细胞凋亡情况;单独应用顺铂组细胞凋亡情况较明显,凋亡率达到(21.27±3.65)%;顺铂联合应用雷帕霉素组细胞凋亡率最高,达到(43.33±5.64)%。雷帕霉素组、顺铂组和顺铂联合应用雷帕霉素组HepG-2细胞均出现点状的自噬泡,顺铂联合应用雷帕霉素组自噬泡数量显著高于雷帕霉素组和顺铂组。结论顺铂可直接诱导HepG-2细胞凋亡,雷帕霉素本身不诱导HepG-2细胞凋亡,但其可显著促进顺铂诱导的HepG-2细胞凋亡;体外侵袭实验结果表明顺铂联合应用雷帕霉素可显著降低HepG-2细胞的体外侵袭能力。
Objective To study the effects of rapamycin combined with cisplatin on the apoptosis, autophagy and cell invasion and metastasis of hepatocellular carcinoma. Methods HepG-2 cells were treated with rapamycin and cisplatin respectively. The inhibition of HepG-2 cell proliferation was detected by MTT assay. The apoptosis of HepG-2 cells was detected by flow cytometry. The transfection chamber was used to determine the invasiveness of HepG-2 cells to the extracellular matrix. Results Compared with the control group, there was no significant apoptosis in the rapamycin alone group. The apoptosis of the cells treated with cisplatin alone was more obvious than that of the control group (21.27 ± 3.65%). The combination of cisplatin Rapamycin group had the highest apoptosis rate (43.33 ± 5.64)%. Rapamycin group, cisplatin group and cisplatin combination of rapamycin group HepG-2 cells were spotted autophagic vacuoles, cisplatin combined with rapamycin group was significantly higher than the number of autophagy bubble ADM group and cisplatin group. Conclusion Cisplatin can directly induce HepG-2 cell apoptosis. Rapamycin does not induce HepG-2 cell apoptosis, but it can significantly promote the apoptosis of HepG-2 cells induced by cisplatin. In vitro invasion assay showed that cisplatin The combination of rapamycin can significantly reduce HepG-2 cells in vitro invasive ability.