索拉非尼对敏感人肝癌细胞株MAPK信号通路基因表达的影响

Gene expression profile of MAPK signal pathway in a human hepatocellular carcinoma cell line sensitive to sorafenib

目的:筛选对索拉非尼敏感的人肝癌细胞株,检测索拉非尼作用于敏感细胞株后MAPK信号通路基因表达谱的变化。方法:索拉非尼作用于人肝癌细胞株Huh7、MHCC97H、HepG2、SMCC7721、HepG2.2.15和PLC,流式细胞术检测肝癌细胞凋亡,CCK-8测定细胞增殖的抑制率,筛选敏感肝癌细胞株;采用实时定量PCR基因芯片,观察索拉非尼作用于敏感细胞株前后MAPK信号通路基因的表达。结果:索拉非尼作用后,流式细胞术的结果显示,凋亡较明显的细胞株为PLC和HepG2.2.15,相对不明显的细胞株为SMCC7721和MHCC97H;CCK-8测定索拉非尼对PLC、HepG2.2.15、Huh7、HepG2、MHCC97H和SMCC7721细胞的IC50值分别为5.25μmol/L、5.30μmol/L、6.80μmol/L、7.01μmol/L、11.7μmol/L和15.0μmol/L。对索拉非尼相对敏感细胞株和不敏感细胞株分别为PLC和SMCC7721。索拉非尼作用敏感细胞株PLC后,MAPK信号通路表达>2.0倍的有2个,≤0.5倍的有6个。结论:PLC为对索拉非尼相对敏感的人肝癌细胞株;索拉非尼主要导致MAPK信号通路中与调控细胞周期和活化转录因子相关的基因表达发生变化。

AIM： To screen a human hepatocellular carcinoma （HCC） cell line that was sensitive to sorafenib and to explore the changes of gene expression profile of MAPK signal pathway in this cell line. METHODS： Human HCC cell lines of Huh7, MHCC97H, HepG2, SMCC7721, HepG2.2.15 and PLC were selected in the study and treated with sorafenib. The apoptotic rate was detected by flow cytometry. The inhibitory effect of sorafenib on cell growth was deter- mined by CCK-8 assay. The changes of gene expression profile of MAPK signal pathway before and after sorafenib treatment in HCC cell lines were measured by real-time gene array. RESULTS： Apoptotic rate was relatively high in PLC and HepG2.2.15 cells, and was relatively low in SMCC7721 and MHCC97H cells. The values of IC50 in PLC, HepG2.2.15, Huh＇/, HepG2, MHCC97H and SMCC7721 cells were 5.25 μmoL/L, 5.30 μmoL/L, 6.80 μmol/L, 7.01 μmol/L, 11.7 μmol/L and 15.0 μmol/L, respectively. The sensitive and resistant HCC cell lines were PLC and SMCC7721, respective- ly. After sorafenib treatment in PLC cells, the gene expression profile of MAPK signal pathway showed that 2 genes were up-regulated to more than 2 times and 6 were down-regulated to below 0.5 times. CONCLUSION： PLC is a sensitive cell line to sorafenib. The changes of gene expression profile after sorafenib treatment are mainly involved in the genes related to cell cycle regulation and transcription factor activation.