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PI3K/AKT/m TOR信号通路的激活能引发人体细胞发生癌变,哺乳动物靶标m TOR作为PI3K/AKT信号通路下游的一个效应分子,被视为针对肿瘤发生和形成的关键治疗靶点,因此阻断该信号通路的相关靶点可以作为恶性肿瘤治疗的一个策略。白细胞介素24(interleukin 24,IL24)是一个选择性诱导肿瘤细胞凋亡的重要抑癌基因。该研究分别利用MTT法和实时无标记细胞功能分析仪检测携带IL24基因的溶瘤腺病毒ZD55-IL24与m TOR抑制剂雷帕霉素(rapamycin)单独或联合作用对多种肝癌细胞的体外杀伤效果;倒置显微镜分别观察ZD55-IL24、雷帕霉素单独作用及两者联合作用引起的细胞形态学变化;Hoechst 33342、流式细胞术和TUNEL染色检测各处理组细胞的凋亡情况;Western blot检测IL24、AKT以及凋亡相关蛋白Bax和Bcl-2的蛋白质水平。结果显示,溶瘤腺病毒ZD55-IL24与雷帕霉素联合作用较两者单独作用更显著地抑制了肝癌细胞Hep3B的生长并诱导了肝癌细胞的凋亡;此外,两者联合作用更有效地上调了肝癌细胞Hep3B中的细胞因子IL24和促凋亡蛋白Bax的表达,同时下调了PI3K/AKT/m TOR信号通路关键蛋白AKT和抗凋亡蛋白Bcl-2的表达。该研究结果表明,雷帕霉素很可能在一定程度上促进了ZD55-IL24病毒介导的IL24表达而增强对肝癌细胞的杀伤作用,进而为肝癌治疗研究提供了一条新型有效的方案。
Activation of the PI3K / AKT / m TOR signaling pathway leads to carcinogenesis in human cells. Mammalian target mTOR, as an effector molecule downstream of the PI3K / AKT signaling pathway, is considered as a key therapeutic target for tumorigenesis and development Disruption of the relevant targets of the signaling pathway may serve as a strategy for the treatment of malignant tumors. Interleukin 24 (IL24) is an important tumor suppressor gene that selectively induces tumor cell apoptosis. In this study, MTT assay and real-time marker-free cell function analyzer were used to detect the in vitro killing of various hepatocarcinoma cells, alone or in combination, with the oncogene adenovirus ZD55-IL24 carrying the IL24 gene and the mTOR inhibitor rapamycin Effects of ZD55-IL24, rapamycin and rapamycin on cell morphology were observed by inverted microscope. The apoptosis of cells in each treatment group was detected by Hoechst 33342, flow cytometry and TUNEL staining. Western blot The protein levels of IL24, AKT and apoptosis-related proteins Bax and Bcl-2 were detected. The results showed that the combined effect of oncolytic adenovirus ZD55-IL24 and rapamycin alone significantly inhibited the growth of Hep3B hepatoma cells and induced the apoptosis of hepatocellular carcinoma cells. In addition, the combined effect of the two drugs was more effective Upregulated the expression of cytokines IL24 and proapoptotic protein Bax in Hep3B hepatocarcinoma cells and down-regulated the expression of key AKT and anti-apoptotic protein Bcl-2 in PI3K / AKT / m TOR signaling pathway. The results show that rapamycin is likely to promote ZD55-IL24 virus-mediated IL24 expression to a certain extent and enhance the killing effect on liver cancer cells, and then provide a new and effective scheme for the treatment of liver cancer.