抑制CIP2A表达对肝癌细胞HepG2生物学特性影响的体内外研究

In vitro and in vivo studies on the effect of inhibiting the expression of CIP2A on the biological characteristics of HepG2 cells

目的探讨抑制CIP2A表达对肝癌细胞HepG2生物学特性的影响。方法运用前期实验已构建成功的重组腺相关病毒载体rAAV2-EGFP-CIP2A shRNA转染肝癌细胞系HepG2,MTT法检测细胞增殖生长情况,流式细胞仪测定细胞凋亡,Transwell小室侵袭实验检测细胞侵袭力。建立裸鼠皮下种植瘤模型,观察裸鼠移植瘤生长,30 d时处死裸鼠并检测肿瘤生长体积及质量,免疫组化法检测移植瘤组织内CIP2A蛋白的表达。结果 MTT及流式细胞仪结果显示,下调CIP2A表达后肝癌细胞HepG2增殖受到抑制,细胞凋亡增加;Transwell小室侵袭实验提示细胞侵袭能力下降。裸鼠皮下种植瘤结果发现,实验干扰组裸鼠皮下种植瘤生长速度、最终体积及质量明显小于空白对照组及rAAV2对照组,移植瘤组织内CIP2A蛋白表达明显降低。结论 CIP2A促进肝癌细胞HepG2增殖、侵袭及体内肿瘤生长。

Objective To investigate the effect of inhibiting CIP2A expression on the biological characteristics of HepG2 cells. Methods The recombinant adeno-associated viral vector rAAV2-EGFP-CIP2A shRNA was successfully transfected into HepG2 cell lines by using previous experiments.Cell proliferation was detected by MTT assay,cell apoptosis was measured by flow cytometry,and cell invasion ability was detected by Transwell assay.A model of subcutaneous implant tumor in nude mice was established to observe the growth of xenograft tumor in nude mice.The volume and quality of tumor growth were detected in nude mice after 30 days,and the expression of CIP2A protein in the transplanted tumor tissues was detected by immunohistochemistry. Results The results of MTT and flow cytometry showed that the proliferation and apoptosis of HepG2 cells were inhibited after downregulation of CIP2A expression.The Transwell invasion experiment indicated a decrease in cell invasion ability.The result of subcutaneously growing tumor in the nude mice found in the interference experiment group of nude mice,the subcutaneous tumor growth rate,the final volume and weight were much smaller than those in blank control group and no-load virus group,CIP2A protein expression in transplanted tumor tissues was significantly lower. Conclusion CIP2A promotes the proliferation,invasion of HepG2 cells and malignant growth in vivo.