PUMA对胃癌SGC-7901细胞增殖的影响

Effect of PUMA on Proliferation of Gastric Cancer SGC-7901 Cells

目的研究外源性p53正向细胞凋亡调控因子(p53up-regulated modulator of apoptosis protein,PUMA)基因对胃癌SGC-7901细胞增殖的影响。方法制备PUMA基因并插入pcDNA 3.1(-)质粒,观察其转染胃癌SGC-7901细胞后PUMA蛋白的表达并用MTT法检测细胞的增殖力[实验设3组,分别为无转染空白胃癌细胞对照(SGC-7901)组,空载质粒转染胃癌细胞对照(SGC-7901-pcDNA3.1)组及重组PUMA质粒实验(SGC-7901-pcDNA3.1-PUMA)组]。结果 PUMA经限制性核酸内切酶切及测序分析,与预期结果吻合。转染质粒后,SGC-7901组与SGC-7901-pcDNA3.1组PUMA表达比较差异无统计学意义(P>0.05,相对表达量分别为0.24±0.01和0.23±0.01),SGC-7901-pcDNA3.1-PUMA组与另2组比较PUMA表达明显增高(P<0.05,相对表达量为0.39±0.01)。SGC-7901、SGC-7901-pcDNA3.1和SGC-7901-pcDNA3.1-PUMA组的A值分别为0.43±0.04、0.46±0.04和0.32±0.03,SGC-7901-pcDNA3.1-PUMA组与另2组比较明显下降(均P<0.01),SGC-7901组与SGC-7901-pcDNA3.1组比较差异无统计学意义(P>0.05)。结论外源性人类PUMA基因对胃癌细胞增殖有抑制作用。

ABSTRACT:Objective To investigate the effect of p53 up-regulated modulator of apoptosis(PU-MA)on the proliferation of gastric cancer SGC-7901 cells.Methods The PUMA gene was cloned and inserted into the eukaryotic expression vector pcDNA 3.1(-).SGC-7901 cells were divided into three groups:no treatment(SGC-7901 group),empty plasmid transfection(SGC-7901-pcD-NA3.1 group),and recombinant PUMA plasmid transfection (SGC-7901-pcDNA3.1-PUMA group).The expression of PUMA protein and the proliferation of SGC-7901 cells were detected after transfection.Results The results of restriction endonuclease digestion and sequencing anal-ysis were consistent with the expected results.The relative expression quantity was 0.24±0.01 in SGC-7901 group,0.23 ± 0.01 in SGC-7901-pcDNA3.1 group and 0.39 ± 0.01 in SGC-7901-pcDNA3.1-PUMA group after plasmid transfection.The A value was 0.43 ±0.04 in SGC-7901 group,0.46±0.04 in SGC-7901-pcDNA3.1 group and 0.32±0.03 in SGC-7901-pcDNA3.1-PU-MA group after plasmid transfection.There were no significant differences in both PUMA ex-pression and A value between SGC-7901 group and SGC-7901-pcDNA3.1 group(P >0.05).Com-pared with SGC-7901 group and SGC-7901-pcDNA3.1 group,PUMA expression increased and A value decreased in SGC-7901-pcDNA3.1-PUMA group(P <0.05 or P <0.01).Conclusion Exog-enous human PUMA gene can inhibit the proliferation of gastric cancer SGC-7901 cells.