甲胎蛋白下调肝癌细胞对奥沙利铂化疗敏感性的作用

Effect of alpha fetoprotein down regulation on chemotherapeutic sensitivity of hepatocellular carcinoma cells to oxaliplatin

目的探讨甲胎蛋白（AFP）下调对肝癌细胞化疗敏感性的研究。方法选取2012-012014-12华北理工大学附属医院收治的经病理诊断确诊的HCC患者56例,采用电化学发光分析法检测肝癌患者化疗前血清中AFP水平;培养正常肝脏细胞L02细胞及肝癌细胞Hep G2细胞,提取细胞蛋白后Western blot检测L02细胞及Hep G2细胞中AFP的表达情况;siRNA技术敲低Hep G2细胞AFP的表达,Western blot检测奥沙利铂诱导敲低AFP表达肝癌细胞的凋亡情况;MTT法测定AFP对Hep G2细胞增殖的影响。结果肝癌患者经奥沙利铂介入化疗未缓解组血清中AFP的水平[（603.60±237.39）ng/ml]明显高于部分缓解组[（482.21±198.99）ng/ml]及完全缓解组[（358.25±143.22）ng/ml]（P〈0.05）。Western blot检测显示在正常肝脏细胞L02中AFP没有表达,而在Hep G2细胞中AFP明显高表达。肝癌细胞Hep G2成功阻断AFP表达后,加入奥沙利铂作用24 h后,Bcl-2的表达量较之Hep G2明显降低,Caspase-3的表达量显著升高;MTT检测显示干扰AFP表达的Hep G2细胞经奥沙利铂作用后的细胞增殖抑制率明显高于奥沙利铂作用后的Hep G2细胞[（71.06±11.22）%vs（50.56±9.98）%,P〈0.05]。结论 AFP可以显著下调肝癌细胞对奥沙利铂化疗的凋亡敏感性和增殖抑制率。

Objective To explore the down-regulation of alpha fetoprotein（ AFP） on the expression of chemotherapeutic sensitivity in hepatocellular carcinoma（ HCC） cells. Methods Fifty-six patients pathologically diagnosed as HCC were collected in the Affiliated Hospital of North China University of Science and Technology. AFP level in serum was detected with Roche Cobas E602 automatic electrochemical luminescence analyzer. The expression of AFP in L02 cells and Hep G2 cells was detected after protein extraction from normal liver L02 cells and hepatocellular carcinoma Hep G2 cells by Western blot. The sensitivity of Hep G2 cells with or without AFP silencing by siRNA-AFP to chemotherapy was detected by Western blot. The apoptosis of AFP in si-AFP Hep G2 cells induced by chemotherapy was tested by Western blot,and the cell proliferation was determined by MTT assay. Results After chemotherapy,AFP level was significantly lower in partial remission group and complete remission group than in non-remission group before chemotherapy [（ 482. 21 ± 198. 99） ng / ml,（ 358. 25 ± 143. 22） ng / ml vs（ 603. 60 ± 237. 39） ng / ml,P 〈0. 05]. Western blot results showed AFP was not expressed in normal liver cells,while it was highly expressed in Hep G2. After oxaliplatin treatment,Bcl-2 expression was significantly decreased in siRNA-AFP Hep G2 cells,while Caspase-3 expression was significantly increased. After oxaliplatin treatment,the proliferation inhibition rate was significantly higher in si-AFP Hep G2 cells than that of Hep G2 cells[（ 71. 06 ± 11. 22） % vs（ 50. 56 ± 9. 98） %,P 〈0. 05）. Conclusion AFP could reduce the apoptosis induced by oxaliplatin and inhibition of cell proliferation in HCC cells.