Dysadherin基因沉默对胰腺癌细胞侵袭移行能力的影响

目的 探讨靶向封闭dysadherin基因对胰腺癌细胞PANC1、BxPC3体外侵袭移行能力的影响.方法 应用脂质体转染方法将小分子RNA(siRNA)转染细胞.实验分为dysadherin-siRNA转染(dysa)组、阴性对照siRNA转染(HK)组、脂质体对照(对照)组、.采用RT-PCR、免疫组化方法检测转染细胞的dysadherin mRNA及蛋白表达;采用Transwell侵袭小室检测转染细胞体外侵袭移行能力.结果 转染dysadherin-siRNA(5 nmol/L)的PANC1和BxPC3细胞的dysadherin mRNA表达较HK组细胞分别下降95.4%、52.1%(P＜0.05);dysadherin蛋白表达亦分别降低91.2%、83.6%(P＜0.01).PANC1细胞的对照组、HK组和dysa组穿膜细胞数分别为163.2±15.5、154.4±17.3和53.6±7.9;BxPC3细胞的对照组、HK组和dysa组穿膜细胞数分别为30.7±3.2、27.5±2.8和4.7±2.4.dysa组显著低于HK组和对照组(P值均＜0.01).结论 应用RNA干扰技术沉默人胰腺癌细胞株PANC1、BxPC3的dysadherin基因可使细胞的侵袭移行能力下降.

Effect of dysadherin gene silencing mediated by RNA interference on metastasis and invasion of pancreatic cancer cells

Objective To investigate the effect of dysadherin gene silencing on metastasis and invasion in pancreatic cancer cell line PANC1,BxPC3 in vitro.Methods PANC1 and BxPC3 cells were divided into dysa group,negative siRNA control group(HK),liposomes control group(control),dysa group and HK group were tranfected by dysadherin siRNA and Negative siRNA,respectively.The expression of dysadherin mRNA and protein were detected by RT-PCR and immunohistochemical method.Transwell test was used to evaluate the invasion ability of pancreatic cancer cells.Results After transfected by dysadherin siRNA,the dysadherin mRNA levels in PANC1 and Bxpc3 cells were decreased by 95.4% and 52.1%.The expression of dysadherin protein was also down-regulated by 91.2% and 83.6%,respectively,when compared with HK groups (P＜0.05 ).The numbers of invasive cells migrated in Transwell in PANC1 cells control group,HK group and dysa group were 163.2±15.5,154.4±17.3 and 53.6±7.9;the numbers of invasive cells in BxPC cells control group,HK group and dysa group were 30.7±3.2,27.5±2.8 and 4.7±2.4,respectively.The numbers in dysa group were significantly lower than those of HK group and control group (P＜0.01 ).Conclusions Silencing the dysadherin gene of PANC1,BxPC3 by RNA interference could significantly inhibit the invasive and migratory ability of canceroas cells.