摘要
【目的】通过检测膀胱癌细胞系中酪氨酸激酶受体C(tyrosine kinase receptor C,TrkC)中的表达水平及小干扰RNA(small interfering RNA,siRNA)干涉膀胱癌细胞中TrkC的表达,分析TrkC在膀胱癌中的作用。【方法】采用RT-PCR和Western blotting的方法检测人膀胱癌细胞株5 637、T24、SCaBER中TrkC的表达,根据TrkC基因序列设计并合成siRNA,构建质粒。用脂质体将p Silencer/TrkC和对照空载体p Silencer转染人膀胱癌细胞株SCaBER,通过Western blotting检测转染细胞的TrkC表达情况,绘制细胞生长曲线,克隆形成实验检测TrkC对SCaBER细胞增殖的影响的影响。【结果】RT-PCR和Western blotting结果均表明人膀胱癌细胞株5 637、T24、SCaBER中TrkC的表达水平明显高于人正常膀胱移行上皮细胞SV-HUC-1;成功利用siRNA下调SCaBER细胞中TrkC的表达后,细胞生长曲线及克隆形成实验结果显示SCaBER细胞出现生长抑制。【结论】TrkC在膀胱癌细胞中的表达高于正常膀胱上皮细胞,下调TrkC表达可以抑制膀胱癌细胞SCaBER增殖,TrkC可能成为膀胱癌基因治疗的新靶点。
【Objective】To analyze the effect of tyrosine kinase receptor C（TrkC） on bladder cancer by detecting TrkC expression levels in different bladder cancer cell lines and down-regulating TrkC expression in bladder cancer cells with siRNA.【Methods】TrkC expression levels in human bladder cancer cell lines（5637, T24 and SCaBER） were detected via RT-PCR and Western blotting.According to TrkC gene sequence, siRNA was designed and synthesized. The related plasmid was constructed. Human bladder cancer cell line SCaBER was transfected by p Silencer/TrkC and p Silencer via Lipofect Amine TM2000 reagent. The TrkC expression in transfected cells was detected with western blot. Cell growth curves were mapped. The effect of TrkC on SCaBER proliferation was detected with clone formation assay.【Results】The results of RT-PCR and western blot indicated that the TrkC expression levels in human bladder cancer lines（5637, T24 and SCaBER） were significantly higher than that in human normal bladder transitional epithelium cell line SV-HUC-1. TrkC expression in SCaBER cells was successfully down-regulated via siRNA. Then cell growth curves and clone formation assay showed that the growth of SCaBER cells was inhibited.【Conclusion】The TrkC expression levels in bladder cancer lines are significantly higher than that in normal bladder epithelium cell line. The proliferation of SCaBER cells can be inhibited by down-regulating TrkC expression. Therefore TrkC may become a new target for gene therapy in bladder cancer.
作者
于新路
于垂恭
YU Xin-lu;YU Chui-gong(Department of Urology,Liaoning Provincial Corps Hospital of PAP,Shenyang 110034,China)
出处
《武警后勤学院学报（医学版）》
CAS
2018年第1期19-23,共5页
Journal of Logistics University of PAP（Medical Sciences）
