沉默RALA基因表达对前列腺癌细胞的增殖、迁移及凋亡的影响  

Effect of Silencing RALA Gene Expression on the Proliferation,Migration and Apoptosis of Prostate Cancer Cells

作　　者：翁吴斌 刘昌明 张家彬 李国敏 薛清平 林宁峰 陈光炳 WENG Wubin;LIU Changming;ZHANG Jiabin;LI Guomin;XUE Qingping;LIN Ningfeng;CHEN Guangbing(Mindong Hospital Affiliated to Fujian Medical University,Fu'an 355000,China;不详)

机构地区：[1]福建医科大学附属闽东医院,福建福安355000

出　　处：《中国医学创新》2023年第7期5-10,共6页Medical Innovation of China

基　　金：福建省自然科学基金项目(2018J01220)。

摘　　要：目的:利用小干扰RNA(siRNA)靶向RALA基因,使其表达受抑制后探讨对前列腺癌细胞增殖、迁移和凋亡的影响及其可能的机制。方法:采用RT-PCR检测前列腺癌DU145、PC-3和LNCap细胞中RALA mRNA的表达水平。通过Lipofectamine^(TM)2000将RALA-siRNA转染体位培养的DU145细胞株,采用MTT法检测RALA-siRNA对DU145细胞株增殖的影响;Transwell实验检测RALA-siRNA对DU145细胞株迁移能力的影响;流式细胞术检测RALA-siRNA对DU145细胞株凋亡影响。应用RTPCR及Western blot方法检测沉默RALA基因后DU145细胞株中RALA的表达水平。结果:RT-PCR检测结果提示DU145细胞株中RALA mRNA的表达水平高于PC-3、LNCap细胞株,其相对表达水平分别为(0.83±0.02)、(0.37±0.07)和(0.41±0.01),差异均有统计学意义(P<0.05)。MTT检测结果显示,转染RALA-siRNA后DU145细胞株的增殖明显受抑制,与空白对照组及阴性对照组比较差异有统计学意义(P<0.05);Transwell实验结果显示,与空白对照组及阴性对照组相比较,RALA-siRNA组穿膜细胞数低于两者(P<0.05)。流式细胞术检测结果显示,RALA-si RNA组细胞凋亡率明显高于空白对照组及阴性对照组(P<0.05)。Western blot检测结果显示比较阴性对照组、空白对照组,RALA-si RNA组的RALA蛋白的表达明显受抑制(P<0.05)。结论:RALA基因在前列腺癌DU145细胞株中的表达水平较高,沉默RALA基因可使DU145细胞增殖、迁移能力受限,提示在前列腺癌治疗上,RALA可作为一个潜在的靶点。Objective:To investigate the effects of small interfering RNA(siRNA)inhibition of RALA gene expression on the proliferation,migration and regulation of prostate cancer cells and its possible mechanisms.Method:RT-PCR was used to detect the expression levels of RALA mRNA,DU145,PC-3 and LNCap cellsin prostate cancer.The RALA-siRNA was transfected into somatic cultured DU145 cell lines by Lipofectamine^(TM)2000,and the effect of RALA-siRNA on the proliferation of DU145 cell lines was detected by MTT assay;the effect of RALA-siRNA on the migration ability of DU145 cell lines was detected by Transwell assay;the flow cytometry was used to assay the effect of RALA-siRNA on apoptosis of DU145 cell line.The expression levels of RALA gene and protein in DU145 cell line after silencing RALA gene were detected by RT-PCR and Western blot.Result:The RT-PCR results suggested that the expression levels of RALA mRNA in DU145 cell line were higher than those in PC-3 and LNCap cell lines(P<0.05),and their relative expression levels were(0.83±0.02),(0.37±0.07)and(0.41±0.01).MTT assay results showed that the expression levels of RALA mRNA after transfection with RALA-siRNA significantly inhibited the proliferation of DU145 cell line,the difference was statistically significant compared with the negative-siRNA and blank control groups(P<0.05);the results of Transwell assay showed that the number of membrane penetrating cells in the RALA-siRNA group was lower than those in the negative-siRNA and blank control groups(P<0.05).The results of flow cytometry showed that the apoptosis rate in the RALA-siRNA group was significantly higher than those in the negative-siRNA and blank control groups(P<0.05).Western blot assay results suggested that the expression of RALA protein in the RALA-siRNA group was significantly inhibited compared with those in the negative-siRNA and blank control groups(P<0.05).Conclusion:The expression level of RALA gene is higher in prostate cancer DU145 cell line,and silencing RALA gene can restrict the proliferatio

关 键 词：RALA 前列腺癌 增殖 凋亡 迁移 小干扰RNA 

分 类 号：R737.25[医药卫生—肿瘤]