重组质粒pEGFP-N1-NPRL2对人胃癌细胞生物学特性的影响  

Effect of recombinant plasmid pEGFP-N1-NPRL2 on the biological characteristics of human gastric cancer cells

作　　者：胡在敏[1] 王川[1] 季文[1] 刘波[1] 蔡璨[1] 姜政[1] 

机构地区：[1]重庆医科大学附属第一医院消化内科,重庆400016

出　　处：《细胞与分子免疫学杂志》2012年第10期1020-1024,共5页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的:探讨pEGFP-N1-NPRL2真核表达载体对SGC-7901胃癌细胞体外增殖﹑细胞周期、凋亡、侵袭及迁移的影响。方法:构建pEGFP-N1-NPRL2真核表达载体,并进行酶切及测序鉴定。脂质体介导转染入SGC-7901胃癌细胞,应用荧光显微镜、RT-PCR、Western blot法检测NPRL2的基因及蛋白水平情况,CCK8法检测细胞增殖的变化,流式细胞仪分析细胞周期和凋亡率的变化,Transwell实验检测NPRL2对细胞迁移及侵袭能力的影响。结果:成功构建了pEGFP-N1-NPRL2真核表达载体。重组质粒转染SGC-7901细胞后,通过荧光显微镜观察到绿色荧光的表达,RT-PCR和Westernblot法检测到NPRL2 mRNA及蛋白的表达,CCK8法检测发现NPRL2能够明显抑制肿瘤细胞增殖(P<0.05),细胞周期分析显示NPRL2使细胞停在G0～G1期(P<0.05),细胞凋亡结果显示NPRL2能抑制细胞凋亡(P<0.05),Transwell侵袭和迁移实验显示NPRL2使细胞侵袭迁移能力均降低(P<0.05)。结论:NPRL2可抑制肿瘤细胞的增殖、周期、侵袭及迁移,并促进凋亡。AIM： To investigate the effect of eukaryotic expression vector pEGFP-N1-NPRL2 on the proliferation, cell cycle, apoptosis, invasion and migration of human gastric carcinoma SGC-7901 cells in vitro. METHODS： The eukaryotic expression vector pEGFP-N1-NPRL2 was con- structed and confirmed by enzyme digestion and sequencing analysis. Then, it was transfected into SGC-7901 cells via the liposome. The expression of NPRL2 mRNA and protein was detected by RT-PCR, fluorescent microscopy and Western blotting, respectively. The proliferation of SGC- 7901 was tested by CCK8, the cell cycle and apoptosis rate by flow cytometry and the effects of NPRL2 on the cell inva- sion and migration by Transwell （ Boyden Chamber） assay. RESULTS： The eukaryotic expression vector pEGFP-N1- NPRL？, was constructed successfully and transfected into SGC-7901cells. The expression of green fluorescent protein was observed using a fluorescence microscope and both mRNA and protein of NPRL2 was detectable by RT-PCR and Western blotting, respectively. CCK8 revealed that the proliferation of SGC-7901 cells were significantly inhibited by NPRL2 （P 〈 0.0.5 ）. Flow cytometry indicated that the cells were arrested at G0/G1 phase （ P 〈 0.0.5 ） and cell apopto- sis was evidently inhibited （ P 〈 0.05 ）. Transwell chamber experiments showed that the abilities of both invasion and migration of the cells decreased by NPRL2 （ P 〈 0. 05 ）. CONCLUSION： The NPRL2 inhibits the proliferation of human gastric carcinoma SGC-7901 cells, arrests cell cycle at GO/G1 phase, decreases the abilities of invasion and migration and promotes apoptosis.

关 键 词：NPRL2 SGC7901 转染 增殖 侵袭 

分 类 号：R735.2[医药卫生—肿瘤] R392.33[医药卫生—临床医学]