慢病毒介导的PRL-3shRNA对结肠癌细胞增殖、侵袭、凋亡的影响  

Effects of sh RNA targeting PRL-3 on proliferation, invasion and apoptosis of human colorectal cancer cells

作　　者：王燕[1] 张姣 王会峰[1] 王宁菊[1] 李国琪 WANG Yan;ZHANG Jiao;WANG Hui-feng;WANG Ning-ju(Department of Medical Oncology, General Hospital of Ningxia Medical University, Yinchuan 750004, China)

机构地区：[1]宁夏医科大学总医院肿瘤内科,750004 [2]不详

出　　处：《天津医药》2018年第3期225-229,共5页Tianjin Medical Journal

基　　金：宁夏自然科学基金资助项目(NZ13147);宁夏医科大学总医院校级课题(XM2012009)

摘　　要：目的观察慢病毒介导的sh RNA沉默肝再生磷酸酶-3(PRL-3)基因对结肠癌SW480细胞增殖、侵袭、凋亡的影响。方法实验分组为空白对照组、阴性对照组、转染组。将携带PRL-3 sh RNA的慢病毒载体转染结肠癌SW480细胞,建立稳定沉默PRL-3的细胞株,real-time PCR检测转染后PRL-3 m RNA的相对表达水平。采用MTT法、平板克隆形成实验检测转染后细胞增殖能力;采用Transwell侵袭实验、侵袭小室法检测转染后细胞迁移及侵袭能力;采用流式细胞术检测转染后细胞凋亡率变化。结果稳定沉默PRL-3的细胞株构建成功,转染组PRL-3m RNA的相对表达水平低于空白对照组、阴性对照组(P<0.05),空白对照组、阴性对照组比较差异无统计学意义。PRL-3 sh RNA转染SW480细胞72 h后,转染组与空白对照组、阴性对照组比较,细胞增殖能力受到抑制,转染120 h时最明显(P<0.05)。转染组克隆形成能力较空白对照组、阴性对照组下降(P<0.05)。转染组与空白对照组、阴性对照组比较,细胞迁移、侵袭能力下降,凋亡率增加(P<0.05)。结论结肠癌SW480细胞转染PRL-3 sh RNA可减少PRL-3的表达,有效抑制SW480细胞增殖,促进其凋亡,PRL-3可能成为治疗结肠癌的靶基因。Objective To investigate the effects of PRL-3shRNA mediated by lentivirus on proliferation,invasion and apoptosis of human colorectal cancer SW480cells.Methods There were three experimental groups in this study,which included blank control group,negative control group and transfected group.Colorectal cancer SW480cells were transfected with lentiviral interference vector carrying PRL-3shRNA to build a stable PRL-3-silencing cell line.The expression of PRL-3mRNA was detected by real-time quantitative polymerase chain reaction(real-time PCR).Cell proliferation was analyzed by MTT method and colony formation assay.Invasion and migration were measured by Transwell assay and invasion chamber.Apoptosis rate was performed by flow cytometry.Results The stable PRL-3-silencing cell line was successfully constructed.Compared with the blank control group and negative control group,the relative expression levels of PRL-3mRNA were reduced in transfected group after transfection with PRL-3shRNA(P<0.05),but there was no significant difference between the blank control group and the negative control group.After transfection with PRL-3shRNA for72h,the proliferation of SW480cells was significantly lower in transfected group than that of the blank control group and the negative control group,and the proliferation decreased significantly in120h(P<0.05).Compared with the blank control group and negative control group,the ability of colony formation was also weakened in the transfected group(P<0.05).Compared with the blank control group and negative control group,the migration and invasion ability were decreased in the transfected group(P<0.05),and the apoptosis rate was increased in the transfected group(P<0.05).Conclusion PRL-3shRNA can inhibit the expression of PRL-3and the proliferation,promote the apoptosis of SW480,which indicates that PRL-3may become a target for colorectal carcinoma treatment.

关 键 词：基因沉默 结肠肿瘤 细胞系 肿瘤 细胞增殖 细胞凋亡 肝再生磷酸酶-3 

分 类 号：R735.3[医药卫生—肿瘤]