重组人颗粒酶B对胃癌细胞增殖的影响

Proliferation analysis of human granzyme Bon stomach carcinoma cells

目的探讨重组人颗粒酶B(G rB)对人胃癌细胞株SGC-7901增殖活性的影响。方法重组表达质粒pCDNA 3.1-G rB经限制性内切酶N otⅠ与X baⅠ双酶切以及DNA测序鉴定无误后,转染处于对数生长期的SGC-7901细胞,同时设空白组、pCDNA 3.1转染组作对照。转染后48h,用RT-PCR法鉴定各组质粒的整合和G rB基因mRNA表达水平;同时采用软琼脂集落形成实验和M TT法检测胃癌细胞生长情况。结果与空白对照组(21.00±2.58)个和pCDNA 3.1转染组(18.50±2.64)个相比,pCDNA 3.1-G rB转染组(9.00±0.82)个的集落形成数目明显减少(P<0.05),细胞生长速度明显降低(P<0.05)。结论G rB对人胃癌细胞增殖有明显抑制作用。

Objective ：To explore the effect of human granzyme B （GrB） on the proliferous activity of human stomach carcinoma cell line SGC-7901. Methods： Recombinant expression plasmid pCDNA3.1-GrB was confirmed by Not I and Xba I double-enzyme incision and DNA sequencing. The right pCDNA3.1-GrB was transfected into SGC-7901 cells by Lipofectamine2000. pCDNA3.1 transfected group and blank transfected group served as controls. 48h after transfection, RT-PCR amplification was used to identify the integration and express of the GrB gene in SGC-7901 cells. Soft agar colony forming experiment and MTT were performed to investigate the effect of GrB on SGC-7901 cells proliferation. Results： Soft agar colony forming method found out that the number of colony formation in pCDNA3.1-GrB group was far less than those in control ,groups. Slower cell grow in pCDNA3.1-GrB group had also been found by MTT. Conclusion： GrB can inhibit the stomach carcinoma cell line-SGC7901.