Department of Oncology, Dongying People's Hospital, Dongying, China.
Department of Oncology, Yantaishan Hospital-Yantai, China.
Asian Pac J Cancer Prev. 2020 Oct 1;21(10):2853-2857. doi: 10.31557/APJCP.2020.21.10.2853.
To explore the effect of combined Sorafenib/ cisplatinum treatment on the autophagy and proliferation of hepatocellular carcinoma (HepG2) cells in vitro.
HepG2 cells were cultured and treated with different concentrations of Sorafenib, cisplatinum, or a combination of both over a 24-hour period. Cell proliferation was evaluated using a CCK8 assay, and the mRNA expression of the autophagy-related proteins AKT, mTOR, and LC3 were detected using quantitative PCR (qPCR). AKT, pAKT (Ser473), mTOR, pmTOR (Ser2448), LC3I, and LC3II protein expression levels were evaluated by western blot.
We found that the survival rate of HepG2 cells was 47.42% when treated with Sorafenib (10 μmol/L) monotherapy, and 46.04% when treated with cisplatinum (10 mg/L) monotherapy. When Sorafenib(10 μmol/L) was combined with cisplatinum (10 mg/L), the cellular proliferation and survival rate was only 16.71% ( P <0.05). qPCR and western blot revealed that a combination of Sorafenib (10 μmol/L) and cisplatinum (10 mg/L) reduced the transcription and protein expression of autophagy-related AKT and mTOR but increased that of LC3 (P <0.05).
Combining Sorafenib and cisplatinum can effectively induce cell autophagy and reduce cellular proliferation via the PI3K/AKT/mTOR signal pathway.
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探讨索拉非尼/顺铂联合治疗对体外肝癌（HepG2）细胞自噬和增殖的影响。
将 HepG2 细胞培养并分别用不同浓度的索拉非尼、顺铂或两者联合处理 24 小时。采用 CCK8 法评估细胞增殖，采用实时定量 PCR（qPCR）检测自噬相关蛋白 AKT、mTOR 和 LC3 的 mRNA 表达。采用 Western blot 法检测 AKT、pAKT（Ser473）、mTOR、pmTOR（Ser2448）、LC3I 和 LC3II 蛋白表达水平。
我们发现索拉非尼（10 μmol/L）单药治疗时 HepG2 细胞的存活率为 47.42%，顺铂（10 mg/L）单药治疗时为 46.04%。当索拉非尼（10 μmol/L）与顺铂（10 mg/L）联合使用时，细胞增殖和存活率仅为 16.71%（P <0.05）。qPCR 和 Western blot 显示，索拉非尼（10 μmol/L）和顺铂（10 mg/L）联合使用降低了自噬相关 AKT 和 mTOR 的转录和蛋白表达，但增加了 LC3 的表达（P <0.05）。
索拉非尼和顺铂联合应用可通过 PI3K/AKT/mTOR 信号通路有效诱导细胞自噬，降低细胞增殖。
