摘要
目的探讨RGC-32基因在胰腺癌Bx PC-3细胞增殖、凋亡中的作用。方法常规培养胰腺癌Bx PC-3细胞,构建RGC-32表达质粒pc DNA3.1/myc-His C-RGC-32,并瞬时转染Bx PC-3细胞作为实验组;将转染空质粒pc DNA3.1/myc-His C的Bx PC-3细胞作为对照组;应用实时定量PCR及Western blotting确定转染效率。采用细胞增殖试剂盒CCK-8(cell counting kit-8)法检测转染前后细胞增殖、流式细胞术检测细胞凋亡的变化。结果定量PCR及Western blotting检测结果均显示转染实验组较对照组RGC-32mRNA及蛋白表达显著上调(P<0.05)。细胞增殖实验表明实验组较对照组细胞存活率显著性下降(P<0.05);细胞凋亡实验表明实验组较对照组细胞凋亡率无显著性下降(P>0.05)。结论在胰腺癌Bx PC-3细胞中,过表达RGC-32可抑制细胞增殖,但对细胞凋亡无明显作用。
Objective To investigate the effect of RGC-32 in cell proliferation and apoptosis of pancreatic cancer cell line BxPC-3.Methods Pancreatic cancer cell line BxPC-3 was routinely cultured.RGC-32 expression plasmid pcDNA3.1/myc-His C-RGC-32 was constructed transiently trasfected into BxPC-3 cells to overexpress RGC-32 and used as experimental group.Emptly vector (pcDNA3.1/myc-His C) was used as a negative control.Quantitative reverse transcription-PCR (qRT-PCR) and western blotting was performed to determine the transfection efficiency.Cell counting kit-8 (CCK-8) and flow cytometry were used to determine the influence of RGC-32 on cell proliferation and apoptosis of BxPC-3 cells respectively.Results QRT-PCR and western blotting demonstrated that the expression level of RGC-32 in experimental group was significantly higher than control group at both mRNA and protein levels.CCK-8 showed that the cell proliferative vitality rate in experimental group was significantly lower than that control group.Cell apoptosis detection showed that there was no difference in apoptosis rate between experimental group and control group.Conclusion RGC-32 overexpression inhibits the proliferation of BxPC-3 cells,but has no obvious effect on cell apoptosis.
出处
《胃肠病学和肝病学杂志》
CAS
2014年第11期1347-1350,共4页
Chinese Journal of Gastroenterology and Hepatology
基金
国家自然科学基金(81302152)
关键词
胰腺癌
RGC-32
增殖
凋亡
Pancreatic cancer
RGC-32
Proliferation
Apoptosis