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目的探讨雷帕霉素(rapamycin)对阿霉素(DOX)治疗肝癌效果的影响以及相关机制。方法 DOX单独或与rapamycin联合处理HepG2、Hep3B 2种肝癌细胞48 h,用MTT法测定细胞的增殖情况;DOX、rapamycin以及两者联合处理HepG2、Hep3B细胞24 h,以流式细胞术检测细胞凋亡率,用Western blot法检测PI3K/Akt/mTOR通路相关蛋白(p-Akt、p-mTOR、p21和p53)的表达。结果 DOX能浓度依赖性抑制HepG2和Hep3B细胞的增殖,且对HepG2细胞的抑制作用明显强于Hep3B细胞;与单用DOX比较,DOX与rapamycin联用对2种细胞增殖的抑制作用明显增加,且增加Hep3B细胞对DOX敏感性的作用尤为显著。DOX作用HepG2和Hep3B细胞24 h,可诱导2种细胞发生不同程度的凋亡,rapamycin单独作用的促凋亡作用不明显,但与DOX联用后能明显增加DOX的促凋亡作用,2种细胞的凋亡率均明显增加。Western blot结果显示,rapamycin能明显上调p-Akt、p21和下调p-mTOR在2种细胞的表达水平,轻度上调p53在HepG2中的表达;DOX能轻度上调p-Akt、p21和下调p-mTOR在2种细胞的表达水平,明显上调p53在HepG2中的表达,Hep3B中无p53表达;两药合用对上述蛋白表达的影响有协同作用。结论 rapamycin能增加DOX对不同p53状态肝癌细胞的增殖抑制和促凋亡作用及肝癌对DOX化疗的敏感性,其机制可能与rapamycin激活凋亡的p53和不依赖p53的p21途径有关。
Objective To investigate the effects of rapamycin on the efficacy of doxorubicin (DOX) in the treatment of liver cancer and the related mechanisms. Methods DOX and Hep3B cells were treated with rapamycin alone or in combination with rapamycin for 48 h. The proliferation of HepG2 and Hep3B cells was determined by MTT assay. The apoptosis of HepG2 and Hep3B cells was detected by flow cytometry (DOX), rapamycin The expressions of PI3K / Akt / mTOR pathway related proteins (p-Akt, p-mTOR, p21 and p53) were detected by Western blot. Results DOX could inhibit the proliferation of HepG2 and Hep3B cells in a concentration-dependent manner, and inhibited the proliferation of HepG2 cells in a dose-dependent manner. Compared with DOX alone, the inhibitory effect of DOX and rapamycin on the proliferation of HepG2 cells was significantly increased The effect of increasing the sensitivity of Hep3B cells to DOX is particularly significant. DOX induced HepG2 and Hep3B cells for 24 h, inducing apoptosis of two kinds of cells with different degrees of apoptosis, while the effect of rapamycin alone was not obvious, but DOX could obviously promote the apoptosis of DOX, and 2 Cell apoptosis rates were significantly increased. Western blot results showed that rapamycin could significantly up-regulate the expression of p-Akt, p21 and down-regulate p-mTOR in both cell lines and slightly upregulate the expression of p53 in HepG2. DOX could up-regulate p-Akt, p21 and down-regulate p The expression of -mTOR in HepG2 and Hep3B cells was significantly up-regulated in the two cell lines, but not in Hep3B cells. The combination of these two drugs had synergistic effects on the expression of these proteins. Conclusions rapamycin can increase the proliferation and apoptosis of hepatoma cells with different p53 status and the sensitivity of DOX to DOX chemotherapy. The possible mechanism is that rapamycin may be involved in p53 activation by rapamycin and p53-independent p21 pathway.