Na^+,K^+-ATP1b1对肝癌细胞生长侵袭能力影响的研究

Proliferation and Invasion Inhibition of Na^+,K^+-ATP1b1 on Hepatocellular Carcinoma Cell

目的:探讨Na+,K+-ATP1b1(ATP1b1)对肝癌细胞生长及侵袭能力的影响。方法:将ATP1b1表达质粒ATP1b1-CMV-FLAG转染肝癌细胞SMMC-7721,以空质粒转染作对照,通过RT-PCR测定ATP1b1的mR-NA表达,在光镜下观察细胞形态学变化,MTT法检测细胞生长增殖,Transwell细胞侵袭实验分析肝癌细胞的侵袭能力。结果:ATP1b1-CMV-FLAG转染后,SMMC-7721的ATP1b1mRNA表达水平明显增高(P<0.05),转染ATP1b1-CMV-FLAG组、pFLAG-CMV-1组和脂质体组的ATP1b1mRNA表达水平分别为1.159、0.182和0.093;转染ATP1b1-CMV-FLAG的SMMC-7721细胞呈变性改变,其生长受到抑制,转染相同DNA浓度的ATP1b1-CMV-FLAG组与pFLAG-CMV-1组对细胞生长的抑制效应有显著性差异(P<0.05);Transwell细胞侵袭实验显示转染ATP1b1-CMV-FLAG组的SMMC-7721细胞的体外侵袭力明显受抑制(P<0.01)。结论:实验结果说明了促进ATP1b1表达能抑制肝癌细胞的生长及侵袭行为。

Objective：To study the effects of Na^＋,K^＋-ATP1b1（ATP1b1）on proliferation and invasion of hepatocellular carcinoma cell.Methods：Hepatocellular carcinoma SMMC-7721 cells were transfected with ATP1b1-CMV-FLAG plasmid expressing ATP1b1 gene,while others were transfected with mock plasmid as negative control.The ATP1b1 mRNA expression levels of cells were measured by RT-PCR and the morphology of cells was observed by light microscope.The proliferation of cells was detected with MTT method and the invasion of cells was analyzed by Transwell cellular invasion test.Results：ATP1b1 mRNA expression in cells transfected with ATP1b1-CMV-FLAG,pFLAG-CMV-1 and Lipofectamine was 1.159,0.182 and 0.093 respectively,and ATP1b1 mRNA expression of SMMC-7721 transfected with ATP1b1-CMV-FLAG was increased markedly（P〈0.05）.The cell morphology was changed and presented cellular degeneration after transfected with ATP1b1-CMV-FLAG.The proliferation of ATP1b1-CMV-FLAG-transfected cells was significantly inhibited（p〈0.05） and positively correlatied to the ATP1b1-CMV-FLAG concentration,while the growth of cells transfected with pFLAG-CMV-1 was inhibited mildly.In Transwell cell ular invasion experiment,the invasion of ATP1b1-CMV-FLAG-transfected cells was notably decreased,comparing to other control groups（P〈0.01）.Conclusion：These results have demonstrated that promoting ATP1b1 expression on hepatocellular carcinoma cell can inhibit its growth and invasion.