中文摘要：

目的 探讨沉默增殖诱导配体（a proliferation-inducing ligand,APRIL）基因对人结肠癌SW480细胞增殖及细胞周期的影响.方法 将APRIL基因的小干扰RNA质粒载体（siRNA-APRIL）转染结肠癌SW480细胞株,以非特异性序列载体转染组（nontargeting control）及未转染组（nontransfected control）作为对照.Real-time PCR和Western blot评价APRIL沉默效率;CCK-8（cell counting kit-8）法检测细胞增殖情况;流式细胞术检测细胞周期变化;RT-PCR检测细胞周期调控基因p21及p27的表达.结果 与非特异性序列载体转染组及未转染组相比,siRNA-APRIL显著抑制APRIL mRNA及蛋白的表达（P＜0.05）;siRNA-APRIL转染SW480细胞48 h、72 h和96 h后细胞增殖能力明显下降（P＜0.05）;转染48 h,siRNA-APRIL组G0/G1期细胞比例增高,S期及G2/M期细胞比例减少,细胞凋亡的数量增加,同时p21及p27 mRNA的表达上调（P＜0.05）;而上述指标两对照组之间比较差异无统计学意义（P＞0.05）.结论 siRNA-APRIL能特异性抑制结肠癌SW480细胞APRIL的表达,并抑制细胞增殖,使细胞周期出现G0/G1期阻滞,其机制可能与上调p21和p27的表达有关.

英文摘要：

Objective To investigate the effects of a proliferation-inducing ligand（APRIL）gene silencing by small interfering RNA（siRNA）on cell cycle and proliferation of colon carcinoma SW480 cells.Methods The siRNA plasmid vector targeting APRIL gene,named as siRNA-APRIL,was transfected into SW480 cells,transfected with scrambled vector as a nontargeting control and nontransfected group as another control.APRIL mRNA and protein expression were examined by real-time PCR and Western blot,respectively.Cell proliferation activity was analyzed by cell counting kit-8（CK-8）,cell cycle was detected by flow cytometry,and p21 together with p27,two important regulatory genes in cell cycle,were measured by RTPCR.Results Compared with nontargeting control and nontransfected control,APRIL expression was inhibited significantly at both mRNA and protein level by siRNA-APRIL being transfected in SW480 cells（P 〈0.05）.Cell proliferation ability was drastically repressed after siRNA-APRIL being transfected at 48 h,72 h and 96 h（P 〈 0.05）.After transfected 48 h,the percent of Go/G1 phase cell was significantly increased,S and G2/M phase cell were significantly decreased,the number of cell in apoptosis was increased and the expression of p21 and p27 mRNA were up-regulated（P 〈 0.05）.There was no significant difference when compared the two control groups each other（P 〉 0.05）.Conclusion siRNA-APRIL can effectively knockdown the expression of APRIL gene in SW480 cells,moreover,it can inhibit the cell proliferation and induce G0/G1 phase cell cycle arrest,which occurrence may involve in upregulation the mRNA expression of p21 and p27.