摘要
目的探讨富含脯氨酸小蛋白2A(small proline-rich protein 2A,SPRR2A)在人胃癌细胞株中的表达水平及其对胃癌细胞增殖和迁移能力的影响。方法用实时荧光定量PCR和Western blot法检测SPRR2A在胃癌细胞中的表达水平;转染靶向SPRR2A的siRNA片段或表达质粒以构建沉默或过表达SPRR2A的胃癌细胞株,并用Western blot法检测转染效率。通过实时无标记细胞分析技术(real time cellular analysis,RTCA)和CCK-8实验以及细胞迁移实验检测SPRR2A对胃癌细胞的增殖和迁移能力的影响。结果SPRR2A mRNA在胃癌细胞株BGC-823、MKN-28和SGC-7901中的相对表达量分别为(849.22±0.04)、(77.17±0.26)和(1.73±0.07),均高于正常胃黏膜细胞GES-1的(1.05±0.06),差异均具有统计学意义(均P<0.05)。SPRR2A蛋白在胃癌细胞株BGC-823、MKN-28和SGC-7901中表达水平均高于正常胃黏膜细胞GES-1(均P<0.05)。在BGC-823细胞中沉默SPRR2A 36 h后,细胞增殖能力和迁移能力均降低(均P<0.05)。在SGC-7901细胞中上调SPRR2A的表达36 h后,细胞增殖能力和迁移能力均增加(均P<0.05)。结论SPRR2A可促进胃癌细胞增殖和迁移能力。
Objective To explore the expression level of small proline-rich protein 2A(SPRR2A)in human gastric cancer cell lines and its effect on the proliferation and migration of gastric cancer cells.Methods Real-time fluorescent quantitative PCR and Western blot were used to detect the expression level of SPRR2A in gastric cancer cells;SPRR2A-targeted siRNA fragments or expression plasmids were transfected to construct gastric cancer cell lines that silence or overexpress SPRR2A,and Western blot was used to detect the transfection effectiveness.The effects of SPRR2A on the proliferation and migration of gastric cancer cells were detected by real time cellular analysis(RTCA),CCK-8 experiments and cell migration experiments.Results The relative expressions of SPRR2A mRNA in gastric cancer cell lines BGC-823,MKN-28 and SGC-7901 were(849.22±0.04),(77.17±0.26)and(1.73±0.07),respectively,which were higher than that of normal gastric mucosal cell line GES-1 which was(1.05±0.06).The differences were all statistically significant(all P<0.05).The expression levels of SPRR2A protein in gastric cancer cell lines BGC-823,MKN-28 and SGC-7901 were higher than that of normal gastric mucosal cell line GES-1(all P<0.05).After silencing SPRR2A in BGC-823 cells for 36 h,cell proliferation and migration ability were reduced(both P<0.05).After up-regulating the expression of SPRR2A in SGC-7901 cells for 36 h,the cell proliferation and migration ability increased(both P<0.05).Conclusion SPRR2A can promote the proliferation and migration of gastric cancer cells.
作者
许晓明
董学妍
陈岳明
余道军
王贤军
Xu Xiaoming;Dong Xueyan;Chen Yueming;Yu Daojun;Wang Xianjun(Department of Pathology,the Second Affi liated Hospital of Zhejiang University School of Medicine,Hangzhou 310009,China;Department of Clinical Laboratory,Affiliated Hangzhou First People’s Hospital of Zhejiang University School of Medicine,Hangzhou 310006,China)
出处
《实用肿瘤杂志》
CAS
2020年第6期511-517,共7页
Journal of Practical Oncology
基金
杭州市医药卫生科技计划项目(2015Z01)
浙江省中医药科技计划项目(2016ZA158)
浙江省医药卫生科技计划(2020KY688)。
