溪黄草对肝癌HepG2细胞基因表达谱的影响

Effect of rabdosia serra (maxim.) hara on gene expression profile of hepatocellular carcinoma HepG2 cells

目的采用表达谱芯片研究溪黄草水提取物对人肝癌细胞HepG2相关基因的影响,探讨溪黄草水提取物对肝癌作用的可能机制。方法体外培养人肝癌细胞HepG2,加入溪黄草水提取物(9.54mg/mL)作用24h后,表达谱芯片检测溪黄草作用后HepG2基因的改变,并用RT-PCR和Western blot验证芯片的结果。结果相差显微镜下观察,发现与阴性对照组相比,溪黄草水提取物作用后HepG2细胞数量明显减少。表达谱芯片结果显示,溪黄草水提取物(9.54mg/mL)作用24h后264个基因比阴性对照组上升2倍以上,194个基因比阴性对照组下降2倍以上。基因本体(GO)和KEGG分析表明溪黄草可上调HepG2细胞DUSPs、IGFBPs家族多个基因,下调MCMs家族多个基因。RT-PCR检测发现与阴性对照组相比,溪黄草处理组DUSP1和IGFBP1升高,FXR和ALDH8A1下降(P<0.01),与表达谱芯片结果一致。Western blot结果发现溪黄草处理组DUSP1蛋白表达也显著高于阴性对照组(P<0.01)。结论表达谱芯片研究表明溪黄草可通过调控多种基因而抑制肝癌细胞增殖。

Objective To adopt the expression profile chip to investigate the effect of Rabdosia serra （Maxim.） hara water extract on related gene of human hepatocellular carcinoma（HCC） HepG2 cells for researching the possible mechanism of its water extract on HCC. Methods The human HCC HepG2 ceils were cultured in vitro. After adding Rabdosia serra （Maxim.） hara water extract （9.54 mg/mL） for 24 h action,the expression profile chip was adopted to detect the HepG2 gene change after Rabdosia ser- ra （Maxim.） hara action and then the chip results were verified by using RT-PCR and Western blot. Results The phase contrast microscope observation found that compared with the negative control group,the number of HepG2 cells was significantly reduced after Rabdosia serra （Maxim.） hara water extract action. The expression profile results showed that after Rabdosia serra （Maxim.） hara water extract action for 24 h, 264 genes were risen to over twice folds compared with the negative control group and 194 genes were decreased by over twice folds compared with the negative control group, The gene ontology（GO） and KEGG analysis indicated that Rabdosia serra （Maxim.） hara could up-regulate multiple genes in DUSPs and IGFBPs family and down-regulate multiple genes in MCMs family. The RT-PCR detection found that compared with the negative control group, DUSP1 and IGFBP1 in the Rabdosia serra （Maxim.） hara treatment group were increased,while FXR and ALDH8A1 were decreased （P〈0. 01） ,which were consistent with the results of expression profile chip. The Western blot results found that the protein expression level of DUSP1 in the Rabdosia serra （Maxim.） hara treatment group was also significantly higher than that in the negative control group （P〈0. 01）. Conclusion The expression profile chip shows that Rabdosia serra （Maxim.） hara can inhibit the proliferation of HCC ceils by regulating multiple genes.