导入PTEN基因抑制胰腺癌细胞体外生长的研究

The in vitro growth suppression of human pancreatic cancer cell line by transfer of the PTEN gene

目的 研究PTEN基因转染对胰腺癌细胞体外生长的生物学效应。方法 以 pBP、pBP PTEN HA、pBP PTENG 12 9R HA质粒分别转染体外培养胰腺癌细胞株JF 30 5 ,并以未转染组为对照 ,Westernblot分析目的基因的表达 ,噻唑蓝比色法 (MTT)检测细胞活力 ,流式细胞术检测细胞凋亡。结果 westernblot示pBP PTEN HA和 pBP PTENG 12 9R HA转染组有HA的表达 ,并分别有PTEN的过表达和PTENG 12 9R的表达 ;MTT示pBP PTEN HA转染组活细胞数较其他各组均低 (pBP PTEN HA组 0 .6 787± 0 .0 785 ;pBP PTENG 12 9R HA组 0 .9847± 0 .10 0 2 ;pBP组1.0 987± 0 .14 80 ;对照组 1.2 0 40± 0 .15 31,P <0 .0 5 ) ,流式细胞术则显示其凋亡率较其他各组明显增高 ( pBP PTEN HA组 11.6 8% ;pBP PTENG 12 9R HA组 4.45 % ;pBP组 3 .5 1% ;对照组 2 .2 7%。P <0 .0 5或P <0 .0 1)。结论 PTEN基因转染可抑制胰腺癌细胞体外生长 ,而这种抑制作用有赖其磷酸酯酶活性并与其促发肿瘤细胞凋亡有关。

Objective To study whether PTEN gene transfer could inhibit the in vitro growth of pancreatic cancer cell line.Methods The plasmids including pBP,pBP-PTEN-HA and pBP-PTEN G-129R-HA were separately transferred into in vitro cultured pancreatic cancer cell line JF-305.The expression of target genes was detected by Western blot.Cell viability was determined by MTT assay.Apoptosis was determined by flow cytometry.Results On immunoblots for PTEN,the band was broader in groups transfected with PTEN or PTEN G-129R than in others,showing overexpression of PTEN and the expression of PTEN G-129R respectively.The expression of exogenous PTEN or PTEN G-129R was also verified by immunoblotting for the HA-tag.MTT assay showed that the group transfected with PTEN had a much lower cell viability than other ones (P<0.05 ).It also had a higher percentage of apoptotic cells than other groups (P<0.05 or P<0.01).Conclusion PTEN gene transfer can suppress the in vitro growth of pancreatic cancer cells,and the effect is dependent upon its phosphotase activity and related to induction of apoptosis.