Huang K, Chen G, Li Y, Liu J K, Wang Z Y, Zhou G C
Department of Urology, Clinical Medical College of Yangzhou University, Yangzhou 225001, China.
Zhonghua Yi Xue Za Zhi. 2016 Nov 1;96(40):3227-3231. doi: 10.3760/cma.j.issn.0376-2491.2016.40.006.
To investigate the expression of p21-activated kinase 1 (PAK1) in bladder cancer and its biological influence on invasion ability of bladder cancer cells. A total of 54 paraffin-embedded bladder cancer tissue samples and 12 normal bladder tissue specimens were retrieved in Jinshan Hospital Affiliated to Fudan University between January 2009 and December 2012. The expression of PAK1 in these tissues was detected by immunohistochemical staining. The PAK1 mRNA and protein levels were measured in bladder cancer cell lines and human normal bladder epithelial cell line using real-time, fluorescence-based quantitative polymerase chain reaction (RT-PCR) and Western blot, respectively. A stable PAK1 gene silencing bladder cancer cell strain 5637 were successfully constructed. After treatment with PAK1 RNA interference(RNAi), the ability of migration and invasion of the 5637 cell was evaluated using a Transwell system. The expression of PAK1 proteins was significantly higher in bladder cancer tissues than in normal bladder tissues (28/54 vs 1/12, <0.05). The overexpression of PAK1 was positively correlated with high histological grade, lymph node metastasis, and tumor size (all <0.05). The mRNA level and protein level of PAK1 was much higher in bladder cancer cell lines T24, 5637 than human normal urothelial cell line SV-HUC-1.PCR and Western blot showed satisfactory inhibitory effect of PAK1 short hairpin RNA (shRNA) on PAK1 expression in 5637 bladder cancer cells. The number of PAK1 RNAi-treated 5637 cells traversed the membrane was decreased compared with the control group in migration and invasion assays. Overexpression of PAK1 in bladder cancer tissues may be an important feature of bladder cancer and related with the metastasis and invasion of bladder cancer. The molecular mechanisms involved in regulation of PAK1 expression needs further research.
探讨p21激活激酶1(PAK1)在膀胱癌中的表达及其对膀胱癌细胞侵袭能力的生物学影响。2009年1月至2012年12月期间,复旦大学附属金山医院共收集了54例石蜡包埋的膀胱癌组织样本和12例正常膀胱组织标本。采用免疫组织化学染色法检测这些组织中PAK1的表达。分别使用实时荧光定量聚合酶链反应(RT-PCR)和蛋白质免疫印迹法检测膀胱癌细胞系和人正常膀胱上皮细胞系中PAK1的mRNA和蛋白质水平。成功构建了稳定的PAK1基因沉默膀胱癌细胞株5637。用PAK1 RNA干扰(RNAi)处理后,使用Transwell系统评估5637细胞的迁移和侵袭能力。PAK1蛋白在膀胱癌组织中的表达明显高于正常膀胱组织(28/54 vs 1/12,<0.05)。PAK1的过表达与高组织学分级、淋巴结转移和肿瘤大小呈正相关(均<0.05)。膀胱癌细胞系T24、5637中PAK1的mRNA水平和蛋白质水平远高于人正常膀胱上皮细胞系SV-HUC-1。PCR和蛋白质免疫印迹显示PAK1短发夹RNA(shRNA)对5637膀胱癌细胞中PAK1的表达有良好的抑制作用。在迁移和侵袭实验中,与对照组相比,PAK1 RNAi处理的5637细胞穿过膜的数量减少。PAK1在膀胱癌组织中的过表达可能是膀胱癌的一个重要特征,并与膀胱癌的转移和侵袭有关。PAK1表达调控的分子机制有待进一步研究。