摘要目的 本次实验探讨分泌型丛生蛋白(secret clusterin,sCLU)调控膀胱癌发生发展的分子机制.方法 本文收集了16对膀胱癌组织样本,通过实时定量聚合酶链反应(RT-PCR)检测sCLU mRNA的表达.小干扰RNA(small interfering RNA,si-RNA)转染膀胱癌细胞中,抑制膀胱癌细胞clusterin基因的表达.CCK-8增殖实验和平板克隆实验用于研究sCLU对膀胱癌细胞增殖的影响.蛋白印迹分析(Western Blot)检测MAPK通路蛋白,如JNK/ERK/P38等通路,研究sCLU对膀胱癌细胞的调控机制.结果 16对膀胱癌组织验证表明sCLU在膀胱癌组织中高表达.敲低膀胱癌细胞中sCLU的表达,细胞增殖能力明显受到抑制(P＜0.05).Western Blot结果显示敲低clusterin基因,P38/MKK3/HSP27信号通路相关蛋白磷酸化水平明显降低.结论 sCLU可能通过P38/MAPK通路调控膀胱癌细胞的增殖能力.

abstractsObjectives To explore the molecular mechanism of sCLU in bladder carcinoma.Methods 16 pairs of bladder were collected as cancer tissues samples,and the expression level of sCLU with PT-PCR was identified.Small interfering RNA (si-RNA) was transfected in bladder carcinoma cell lines to knock down the expression of gene clusterin.Cell proliferation and colony formation assay were done to evaluate the changes on the biological characters of tumor cells.Western blot was used to determine the changes of MAKPs signaling,such as JNK,ERK and P38.Results sCLU had a high level expression in human bladder carcinoma tissues.Knocking down the gene clusterin,the bladder carcinoma cell lines had a lower cell proliferation abilities (P ＜ 0.05).Western blot showed that knocking down the clusterin expression declined the phosphorylation levels of P38/MKK3/HSP27.Conclusions Silence of gene clusterin effectively inhibits the proliferation through MAPK/P38 signal pathway.