Wu Yan, Cai Qin, Li Wei, Cai Zhensheng, Liu Ying, Li Hongfan, Pang Ji, Chen Yongchang
Department of Physiology, School of Medicine, Jiangsu University, Zhenjiang 212013, China
The Fourth Affiliated Hospital of Jiangsu University, Zhenjiang 212001, China.
Biosci Rep. 2019 Aug 13;39(8). doi: 10.1042/BSR20190405. Print 2019 Aug 30.
Despite advances in chemotherapy, ovarian cancer (OC) is still the most lethal gynecologic malignancy. So, it is imperative to explore its mechanism and find novel targets to improve the outcome. Type II cyclic guanosine 3',5'-monophosphate (cGMP)-dependent protein kinase (PKG II) has been recently reported to inhibit proliferation and metastasis in several tumors. The present study is to clarify the effect of PKG II combined with l-arginine (l-Arg) on OC cells. SKOV3 and A2780 cells were infected with adenovirus coding cDNA of PKG II to increase PKG II expression and l-Arg was applied to activate this kinase. CCK8 assay, Transwell migration and TUNEL assay were applied to detect the proliferation, migration and apoptosis of the OC cells, respectively. Western blotting was used to detect the level of total and phosphorylated proteins. Our results showed that co-treatment with PKG II and l-Arg inhibited EGF-induced proliferation and the expression of Proliferating Cell Nuclear Antigen (PCNA), Cyclin E and N-Cadherin, whereas up-regulated the expression of E-Cadherin, abolished the anti-apoptotic effect of EGF, prevented the process of epithelial-to-mesenchymal transition (EMT) as well as blocked EGF-triggered Raf-MEK and phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathways. Our results suggested that PKG II activated by l-Arg could inhibit proliferation and migration and promote the apoptosis of OC cells. Based on the above results and our previous data, it is speculated that PKG II is an inhibitor of cancer with extensive effects.
尽管化疗取得了进展，但卵巢癌（OC）仍然是最致命的妇科恶性肿瘤。因此，探索其发病机制并寻找新的靶点以改善治疗效果势在必行。最近有报道称，II型环磷酸鸟苷（cGMP）依赖性蛋白激酶（PKG II）可抑制多种肿瘤的增殖和转移。本研究旨在阐明PKG II与L-精氨酸（L-Arg）联合应用对OC细胞的影响。用编码PKG II cDNA的腺病毒感染SKOV3和A2780细胞以增加PKG II的表达，并应用L-Arg激活该激酶。分别采用CCK8法、Transwell迁移实验和TUNEL实验检测OC细胞的增殖、迁移和凋亡情况。采用蛋白质免疫印迹法检测总蛋白和磷酸化蛋白水平。我们的结果表明，PKG II与L-Arg联合处理可抑制表皮生长因子（EGF）诱导的增殖以及增殖细胞核抗原（PCNA）、细胞周期蛋白E和N-钙黏蛋白的表达，而上调E-钙黏蛋白的表达，消除EGF的抗凋亡作用，阻止上皮-间质转化（EMT）过程，并阻断EGF触发的Raf-丝裂原活化蛋白激酶/细胞外信号调节激酶（MEK）和磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）信号通路。我们的结果提示，L-Arg激活的PKG II可抑制OC细胞的增殖和迁移并促进其凋亡。基于上述结果和我们之前的数据推测，PKG II是一种具有广泛作用的癌症抑制剂。
