上调XB130表达对人肝癌HepG2细胞凋亡和增殖的影响

目的观察XB130 表达上调后人肝癌细胞HepG2 增殖和凋亡的变化，并探讨其作用机制。方法Western blot及实时荧光定量聚合酶链反应（qRT-PCR）检测肝癌细胞株中XB130表达；构建过表达质粒pCMVEGFP-XB130 转染HepG2 细胞，实验分为pCMV-EGFP-XB130 组（pCMV-EGFP-XB130 转染），空白对照组（EGFP 转染）。以免疫荧光、Western blot法及qRT-PCR 法检测上调效率。Western blot 法检测PI3K/Akt通路相关基因表达；采用MTT法检测转染后细胞增殖，流式细胞术检测细胞凋亡。结果XB130 蛋白及其mRNA在肝癌细胞株Hep3B、Huh7、HepG2 和MHCC97H 中均有表达，在HepG2 细胞株中表达量最低（ F=6.342 和5.424，均P=0.001）。与EGFP 组比较，pCMV-EGFP-XB130 组中p-p21、p-p27、p-FOXO3a 及Pro Capase-9蛋白表达上调（t =4.652，3.558，6.743 和3.021，均P<0.05）；XB130 表达上调后在72 及96 h HepG2 细胞增殖能力增强（ t=4.752 和8.542，均P=0.001）；XB130 表达上调后HepG2 细胞凋亡率降低（t =7.467，P =0.001）。结论XB130 表达上调通过PI3K/Akt信号通路促进肝细胞癌细胞增殖，抑制凋亡。

Effect of XB130 on human hepatocellular carcinoma HepG2 cells

Objective To investigate the effect of XB130 on proliferation and apoptosis of human hepatocellular carcinoma HepG2 cell lines. Methods Western blot and qRT-PCR were utilized to determine expression of XB130 in hepatocellular carcinoma cells. Over expressed plasmid pCMV-EGFP-XB130 was transfected into HepG2 cells. Expression of XB130 was identified by Immunofluorescence, Western blot, and qRT-PCR. PI3K/Akt pathway related proteins were measured by Western blot; proliferation and apoptosis rate were measured by MTT assay and Flow cytometry, respectively. Results XB130 protein and mRNA were detected in different hepatocellular carcinoma cell-lines including Hep3B, Huh7, HepG2 and MHCC97H while HepG2 showed the lowest concentration of XB130 (P = 0.001). p-p21, p-p27, p-FOXO3a and pro-Caspase 9 protein in pCMV-EGFP-XB130 group was up-regulated significantly when compared with EGFP control group(t = 4.652, 3.021, 3.558 and 6.743, P< 0.05); XB130 overexpression increased cellular proliferation (P = 0.001) while decreased apoptosis at the 72nd and 96th ( P= 0.001) in HepG2. Conclusion Up-regulation of XB130 increases proliferation while inhibits apoptosis via modulating PI3K/Akt signal pathway in hepatocellular carcinoma HepG2.