摘要
目的探讨DDX5基因表达下调对胃癌细胞增殖、迁移和侵袭的影响。方法采用慢病毒介导的DDX5-shRNA下调人胃癌HGC-27细胞中DDX5的表达。实时荧光定量PCR和蛋白免疫印迹(Western blot)法验证下调效果。CCK8和平板克隆检测细胞增殖能力,Transwell检测细胞迁移和侵袭能力的改变。结果 DDX5-shRNA可有效下调人胃癌HGC-27细胞中DDX5的mRNA和蛋白质表达。DDX5表达下调后,人胃癌HGC-27细胞的增殖受到抑制,24、48、72 h细胞增殖能力较转染了control-shRNA的细胞分别降低25.0%、33.3%和44.6%;转染了control-shRNA的细胞和转染了DDX5-shRNA的细胞克隆形成的数目分别为(162.6±10.3)个和(68.7±11.2)个,两组间比较,差异有统计学意义(P<0.05);转染了control-shRNA的细胞和转染了DDX5-shRNA的细胞24 h迁移数目分别为(153.7±9.2)个和(78.0±6.7)个,两组间比较,差异有统计学意义(P<0.05);转染了control-shRNA的细胞和转染了DDX5-shRNA的细胞24 h侵袭数目分别为(83.7±12.8)个和(40.2±7.6)个,两组间比较,差异有统计学意义(P<0.05)。结论 DDX5表达下调可明显抑制人胃癌HGC-27细胞的增殖、迁移及侵袭,其有可能成为胃癌的治疗靶点。
Objective To investigate the effects of DDX5 down- regulation on gastric cancer cell proliferation,migration and invasion. Methods Lentivirus mediated shRNA were used to down-regulate the expression of DDX5 in human gastric cancer HGC-27 cells. Real-time fluorescent quantitative PCR and Western blot method were used to verify the down-regulation effect. CCK8 and colony formation assay were employed to examine cell proliferation. Transwell assay was performed to detect cell migration and invasion.Results DDX5-shRNA effectively inhibited both mRNA and protein expression of DDX5. DDX5-shRNA significantly suppressed cell proliferation. Cell proliferation was inhibited by 25. 0%,33. 3% and 44. 6% in 24,48,72 hours,compared with control-shRNA cells.Moreover,colony formation was also inhibited. The number of colonies in 24 hours was( 162. 6 ± 10. 3) and( 68. 7 ± 11. 2) in the control-shRNA and DDX5-shRNA group,respectively,the difference between the two groups had statistical significance( P〈0. 05). Furthermore,cell migration was also inhibited by DDX5 down-regulation. The number of migrated cells in 24 hours was( 153. 7 ± 9. 2) and( 78. 0 ± 6. 7) in the control-shRNA and DDX5-shRNA groups,respectively,the difference between the two groups had statistical significance( P〈0. 05). Finally,cell invasion was also reduced. The number of invaded cells in 24 hours was( 83. 7 ± 12. 8) and( 40. 2 ±7. 6) in the control and DDX5-shRNA groups,respectively,the difference between the two groups had statistically significance( P〈0. 05). Conclusion Down-regulation of DDX5 significantly inhibits the proliferation,migration and invasion of gastric cancer HGC-27 cells.
出处
《临床军医杂志》
CAS
2017年第1期16-19,23,共5页
Clinical Journal of Medical Officers
基金
北京医卫健康公益基金(YWKYQ2001)
军队医学科技青年培育计划(15QNP005)
关键词
DDX5
胃癌
增殖
迁移
侵袭
DDX5
Gastric cancer
Proliferation
Migration
Invasion