摘要目的 观察小干扰RNA(siRNA)抑制REG1A(REG1A)基因的表达对人膀胱癌EJ细胞增殖及侵袭的影响.方法 化学合成针对REG1A基因的3对siRNA,使用脂质体转染EJ细胞.实时定量聚合酶链反应( Real-time PCR)和免疫印迹(Western blot)法分别检测转染细胞REG1A的mRNA及蛋白表达,细胞计数试剂盒(CCK-8)法检测转染细胞的增殖,流式细胞术检测转染细胞的周期,Transwell侵袭小室检测转染细胞的侵袭能力.结果 REG1A siRNA转染组EJ细胞与空白对照Mock组比较,REG1A的mRNA及蛋白表达明显下调,分别下调了(75.58±1.17)％及(51.22±6.63)％ (P ＜0.01)；REG1A siRNA转染组EJ细胞增殖速度较空白对照Mock组及阴性对照NC组明显减慢(P＜0.05),且G0/G1期细胞百分比明显增多,为(43.37±2.15)％(P＜0.01)；REG1AsiRNA转染组EJ细胞穿过Transwell小室的数目为(95.84±6.49)个,明显少于空白对照Mock组的(179.93 ±8.38)个和阴性对照NC组的(186.96±6.28)个(P＜0.01).结论 REG1A siRNA能抑制体外培养的膀胱癌EJ细胞增殖并降低其侵袭能力.

abstractsObjective To explore the effects of silencing regenerating islet-derived 1 alpha (REG1A) gene by small interfering RNA (siRNA) on cell growth and invasion of human bladder cancer EJ cells.Methods Three small interfering RNAs targeting human REG1A mRNA were transfected into EJ cells using a liposome approach.Real-time quantitative polymerase chain reaction (RT-PCR) and Western blotting were used to examine REG1A gene and protein expression in EJ cells respectively.Cell proliferation and cell cycle were evaluated by cell counting Kit-8 (CCK-8) method and flow cytometry,and cell invasion was detected using Transwell chambers.Results The expression of REG1A mRNA level and prorein level in the REG1A siRNA transfected cells was significantly down-regulated (75.58 ± 1.17)％ and (51.22 ± 6.63 ) ％ compared with that of the mock cells respectively ( P ＜ 0.01 ).Cell proliferation was significantly decreased in the REG1A siRNA transfected cells compared with that of the mock and negative control cells (P ＜ 0.01 ).REG1A siRNA caused a striking G0/G1 arrest (P ＜ 0.01 ).The proportion of G0/G1 phase cells was (43.37 ±2.15)％ in the REG1A siRNA transfected cells.The number of cells invading through the Transwell chambers in the REG1A siRNA transfected cells (95.84 ± 6.49 ) was significantly lower than those in the mock cells ( 179.93 ± 8.38) and negative cells ( 186.96 ± 6.28 ) ( P ＜0.01 ).Conclusion Silencing of REG1A inhibits cell proliferation and decreases invasion ability of EJ cells in vitro.