论文部分内容阅读
目的 探讨富含亮氨酸重复序列免疫球蛋白样蛋白1(LRIG1)基因表达下调后对膀胱癌T24细胞增殖和凋亡的影响,并探讨其可能的作用机制.方法 利用LRIG1 siRNA转染人膀胱癌T24细胞,并应用RT-PCR和Western blot技术检测转染LRIG1 siRNA后膀胱癌T24细胞中LRIG1的表达.利用CCK 8试剂盒和流式细胞术分别检测下调LRIG1基因的表达对T24细胞增殖和凋亡的影响,进一步应用RT-PCR和Western blot检测与增殖和凋亡相关的Bax和Bcl-2基因的表达.结果 转染LRIG1 siRNA后,LRIG1基因在膀胱癌124细胞中的表达明显下调(P＜0.05).与未处理组和对照组相比,实验组在转染24、48、72、96 h后T24细胞的增殖率均显著高于未处理组和对照组(96h:2.621 ±0.059 vs 2.145 ±0.066,2.201 ±0.064;F=51.049,P=0.000).转染48 h后,实验组细胞早期凋亡率为(5.713 ±0.761)％,显著低于未处理组和对照组的早期凋亡率[(8.412 ±0.848)％,(8.631 ±0.715)％;F=13.137,P=0.006].此外,LRIG1表达的下调能引起Bax基因表达的降低(P＜0.05),Bcl-2基因表达的升高(P＜0.05).结论 LRIG1基因可能在膀胱癌T24细胞的增殖和凋亡中发挥重要作用.“,”Objective To investigate the effects of down-regulation of leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) expression on cell proliferation and apoptosis in bladder cancer cell line T24 and to explore the possible molecular mechanisms involved.Methods Small interfering RNA (siRNA) against the LRIG1 gene was transfected into T24 cells using LipofectamineTM 2000.After transfection,LRIG1 mRNA and protein expressions were detected by reverse transcription and polymerase chain reaction (RT-PCR) and Western blot,res pectively.Cell proliferation was determined using cell counting kit8 (CCK-8).Cell apoptosis was measured by flow cytometry.Furthermore,the mRNA and protein expressions of Bax and Bcl-2 were detected by RT-PCR and Western blot,respectively.Results LRIG1 mRNA and protein expressions were significantly down-regulated after transfection with LRIG1 siRNA (P ＜0.05).Compared to untransfected cells and cells transfected with control siRNA,the proliferation of T24 cells was significantly faster after transfection with LRIG1 siRNA for 24,48,72,and 96 h (96 h:2.621 ±0.059 vs 2.145 ±0.066,2.201 ±0.064; F =51.049,P =0.000),and the early apoptosis rate of T24 cells transfected with LRIG1 siRNA was significantly lower [(5.713 ± 0.761) ％ vs (8.412 ± 0.848) ％,(8.631 ± 0.715) ％ ; F =13.137,P =0.006].In addition,LRIG1-knockdown down-regulated the Bax expression,but up-regulated the Bcl-2 expression in T24 cells (all P ＜ 0.05).Conclusions LRIG1 plays an important role in the proliferation and apoptosis of bladder cancer T24 cells.