Guan Wen-Ying, Zhao Sheng, Luo Yun-Na
Myopia Laser Treatment Center, Affiliated Hospital of Inner Mongolia Medical University, Hohhot, China.
Department of Image Diagnostics, Affiliated Hospital of Inner Mongolia Medical University, Hohhot, China.
Cell Biol Int. 2020 Oct;44(10):2107-2119. doi: 10.1002/cbin.11419. Epub 2020 Jul 24.
This study aims to investigate the expression of retinoblastoma binding protein 6 (RBBP6) in prostate cancer (PCa) and its association with the c-Jun N-terminal kinase (JNK) pathway. Immunohistochemistry was used to detect RBBP6 and JNK1/2 expression in PCa and benign prostatic hyperplasia tissues. RBBP6 expression in PCa cells (LNCap, PC3, and DU145) and noncancerous prostate epithelial cells (RWPE-1) was determined by quantitative real-time polymerase chain reaction and western blot analysis. PC3 and DU145 cells were transfected with RBBP6 small interfering RNAs (siRNAs) to examine the biological characteristics. Anisomycin (a JNK activator) with/without RBBP6 siRNA was used to treat PC3 cells for further investigating the ramification of the RBBP6-mediated JNK pathway in PCa. PCa tissues and cells showed higher RBBP6 and JNK1/2 expression. RBBP6 was positively correlated with JNK1/2 in PCa tissues. Besides, RBBP6 expression was correlated to clinical tumor stage, lymph node metastasis, Gleason grade, preoperative prostate-specific antigen level, as well as prognosis of PCa. RBBP6 siRNA reduced cell proliferation, arrested cells at G2/M, and promoted cell apoptosis, and suppressed JNK pathway. In addition, migration and invasion decreased after the RBBP6 siRNA transfection with downregulated matrix metallopeptidase-2 (MMP-2) and MMP-9. Anisomycin promoted the proliferation, invasion, and migration of PC3 cells and inhibited PC3 cell apoptosis, which could be reversed by RBBP6 siRNA. RBBP6 expression was upregulated in PCa tissues and positively correlated with expression level of JNK1/2. With inhibition of RBBP6 expression, the proliferation, invasion, and migration of PCa cells decreased dramatically, while PC3 cell apoptosis increased appreciably, accompanied by the suppression of the JNK pathway.
本研究旨在探讨视网膜母细胞瘤结合蛋白6（RBBP6）在前列腺癌（PCa）中的表达及其与c-Jun氨基末端激酶（JNK）通路的关联。采用免疫组织化学法检测PCa和良性前列腺增生组织中RBBP6和JNK1/2的表达。通过定量实时聚合酶链反应和蛋白质印迹分析确定PCa细胞（LNCap、PC3和DU145）及非癌前列腺上皮细胞（RWPE-1）中RBBP6的表达。用RBBP6小干扰RNA（siRNAs）转染PC3和DU145细胞以检测其生物学特性。用茴香霉素（一种JNK激活剂）联合或不联合RBBP6 siRNA处理PC3细胞，以进一步研究PCa中RBBP6介导的JNK通路的分支情况。PCa组织和细胞中RBBP6和JNK1/2表达较高。在PCa组织中，RBBP6与JNK1/2呈正相关。此外，RBBP6表达与临床肿瘤分期、淋巴结转移、Gleason分级、术前前列腺特异性抗原水平以及PCa的预后相关。RBBP6 siRNA可降低细胞增殖，使细胞停滞于G2/M期，促进细胞凋亡，并抑制JNK通路。此外，RBBP6 siRNA转染后，基质金属蛋白酶-2（MMP-2）和MMP-9表达下调，细胞迁移和侵袭能力降低。茴香霉素可促进PC3细胞的增殖、侵袭和迁移，并抑制PC3细胞凋亡，而RBBP6 siRNA可逆转这种作用。PCa组织中RBBP6表达上调，且与JNK1/2表达水平呈正相关。抑制RBBP6表达后，PCa细胞的增殖、侵袭和迁移能力显著降低，而PC3细胞凋亡明显增加，同时JNK通路受到抑制。
