摘要
目的应用慢病毒转染技术研究αB-晶状体蛋白(CRYAB)基因沉默对人胃癌SGC-7901细胞增殖和迁移能力的影响。方法构建CRYAB慢病毒载体转染人胃癌细胞株SGC-7901,获得稳定转染的细胞株,制备空载体作为阴性对照组(sh-ctrl组),实验组为CRYAB病毒转染组(sh-CRYAB组)。提取总RNA和蛋白,应用荧光定量PCR和蛋白免疫印迹法检测CRYAB的表达,应用CCK-8、Transwell和细胞划痕实验分别检测CRYAB基因沉默后细胞增殖和迁移能力的变化。结果慢病毒转染SGC-7901细胞后,CRYAB mRNA及蛋白水平显著降低(P<0.01,P<0.001),表明成功构建了稳定转染的CRYAB-shRNA胃癌细胞株。与sh-ctrl组比较,sh-CRYAB组细胞的增殖能力在第4天、第5天显著降低(P<0.05,P<0.01),sh-CRYAB组细胞迁移率显著降低(P<0.01)。结论 CRYAB表达沉默可降低胃癌细胞的增殖和迁移能力,CRYAB有望成为胃癌基因治疗的新靶点。
Objective The effect of silencing alpha B-crystallin （CRYAB） on cell proliferation and migration of human gastric cancer cell line SGC-7901 was investigated by lentiviral transfection. Methods A lentiviral mediated CRYAB-shRNA was constructed and successfully transfected into SGC-7901 cell. The empty vector was used as the negative control group （sh-ctrl group）, the experimental group was CRYAB virus transfection group （sh-CRYAB group）.The expression of CRYAB on mRNA and protein level was detected by Real-time PCR and Westem blot, respectively. Cell proliferation was detected by CCK8 assay. The cell migration was analyzed by Transwell migration assay and cell scratch test. Results After lentivirus was transfected into SGC- 7901 cells, the expression of CRYAB gene was decreased on mRNA and the protein level significantly （P 〈 0.01, P 〈 0.001）. A stable transfected CRYAB-shRNA gastric cancer cell line was successfully constructed. The cell proliferation rate of sh-CRYAB group was significantly lower （P 〈 0.05, P 〈 0.01）, and the cell mi- gration rate of sh-CRYAB group was significantly lower than sh-ctrl group （P 〈 0.01）. Conclusion Silencing CRYAB expression might effectively inhibit cell proliferation and migration in SGC-7901 cell. CRYAB gene might be a promising novel target in gene therapy of gastric cancer.
出处
《兰州大学学报（医学版）》
CAS
2017年第2期1-6,共6页
Journal of Lanzhou University（Medical Sciences）
基金
甘肃省科技支撑计划项目(1204FKCA172)
中央高校基本科研业务费专项资金项目(lzujbky-2016-59
lzujbky-2016-62)
关键词
αB-晶状体蛋白基因
慢病毒
细胞增殖
细胞迁移
胃癌
alpha B-crystallin gene
lentivirus transfection
proliferation
migration
gastric cancer
