Zhan Bo, Kong Chuize, Zhang Zhe, Dong Xiao, Zhang Naiwen
Department of Urology, The First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.
Exp Ther Med. 2017 May;13(5):2473-2479. doi: 10.3892/etm.2017.4258. Epub 2017 Mar 23.
Kidney cancer is among the most important causes of cancer-associated mortality worldwide. The present study aimed to evaluate protein kinase C α (PKCα) expression in kidney cancer tissues and cell lines, and its significance in apoptosis and migration. Expression of PKCα was analyzed using quantitative polymerase chain reaction and western blotting. In addition, the inhibitor of PKCα (calphostin C and GO6976) was used to treat kidney cancer cells. The ACHN cell line was generated with PKCα-small-interfering RNA (siRNA) and a stable expression of PKCα, in order to facilitate the analysis of apoptosis and migration of PKCα during knockdown and inactivation. Flow cytometry was used to determine the rates of apoptosis. Immunohistochemical staining was used to identify the localization of PKCα in renal clear cell carcinoma and normal sections. PKCα expression in normal tissues was found to be greater than in cancerous tissues. Furthermore, apoptosis was not promoted with PKCα inhibitors or PKCα-siRNA treatment, and a decrease of the migration ability was observed following transfection with PKCα-dominant negative. The results indicated that inhibition of PKCα might not contribute to apoptosis progression in kidney carcinoma.
肾癌是全球癌症相关死亡的最重要原因之一。本研究旨在评估蛋白激酶Cα(PKCα)在肾癌组织和细胞系中的表达及其在细胞凋亡和迁移中的意义。采用定量聚合酶链反应和蛋白质印迹法分析PKCα的表达。此外,使用PKCα抑制剂(钙泊三醇C和GO6976)处理肾癌细胞。用PKCα小干扰RNA(siRNA)构建ACHN细胞系并使其稳定表达PKCα,以便于分析PKCα在敲低和失活过程中的细胞凋亡和迁移情况。采用流式细胞术测定细胞凋亡率。免疫组织化学染色用于鉴定PKCα在肾透明细胞癌和正常切片中的定位。发现PKCα在正常组织中的表达高于癌组织。此外,PKCα抑制剂或PKCα-siRNA处理并未促进细胞凋亡,而转染PKCα显性阴性后观察到迁移能力下降。结果表明,抑制PKCα可能无助于肾癌细胞凋亡的进展。