Promotion of Cell Proliferation through Inhibition of Cell Autophagy Signalling Pathway by Rab3IP is Restrained by MicroRNA-532-3p in Gastric Cancer
Promotion of Cell Proliferation through Inhibition of Cell Autophagy Signalling Pathway by Rab3IP is Restrained by MicroRNA-532-3p in Gastric Cancer
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胃癌中 MicroRNA-532-3p 抑制 Rab3IP 通过抑制细胞自噬信号通路促进细胞增殖
DOI：
10.7150/jca.27533
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发表时间：
2018-01-01
影响因子：
3.9
通讯作者：
Hu, Yanfeng
中科院分区：
文献类型：
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作者：
Guo, Weihong;Chen, Zhaoyu;Hu, Yanfeng
背景：rab3a相互作用蛋白（Rab3IP）被认为与癌症有关；然而，其在胃癌（GC）细胞增殖过程中的功能尚不清楚。因此，本研究旨在探讨Rab3IP在GC中的潜在功能。方法：采用实时荧光定量PCR和免疫组化检测Rab3IP的表达及其临床病理价值。将Rab3IP（敲低和过表达）和轻链3 （LC3）慢病毒转染GC细胞，通过细胞计数试剂盒-8、平板克隆形成、流式细胞术和肿瘤发生试验检测细胞增殖情况。采用共聚焦激光扫描显微镜和免疫印迹法检测细胞自噬。荧光素酶报告基因法分析microRNA-532-3p （miR-532-3p）对Rab3IP的调控。结果：GC样品中Rab3IP的过表达增强了细胞的增殖能力，但减少了自噬体的数量和LC3-II和sequestoome -1 （SQSTM1或p62）标记物的表达。此外，我们发现miR-532-3p可以结合RAB3IP的3′UTR区，抑制GC细胞的增殖能力。miR-532-3p的表达与Rab3IP的表达呈负相关。结论：我们的研究阐明了Rab3IP通过参与自噬在诱导胃癌细胞增殖中的核心作用。miR-532-3p直接靶向Rab3IP并抑制其功能，从而在GC中展示了一种新的调控联系。
Background: RAB3A-interacting protein (Rab3IP) is known to be involved in cancer; however, its function during the proliferation of gastric cancer (GC) cells remains unknown. Therefore, this study aimed to explore the potential function of Rab3IP in GC. Methods: The expression of Rab3IP and its clinical pathology value were determined by quantitative real-time PCR and immunohistochemistry. Rab3IP (knockdown and overexpression) and light chain 3 (LC3) lentiviruses were transfected into GC cells, and cell proliferation was measured using cell counting kit-8, plate clone formation, flow cytometry, and tumorigenesis assays. Cell autophagy was measured using a confocal laser scanning microscope and by western blotting. Luciferase reporter assay was performed to analyse the regulation of Rab3IP by microRNA-532-3p (miR-532-3p). Results: Overexpression of Rab3IP in GC samples enhanced the cell proliferation ability, but decreased the number of autophagosomes and expression of LC3-II and sequestosome-1 (SQSTM1 or p62) markers. Furthermore, we found that miR-532-3p can bind to the 3ʹUTR region of RAB3IP and inhibit the proliferation ability of GC cells. Further, the expression of miR-532-3p negatively correlated with that of Rab3IP. Conclusions: Our study elucidates the central role of Rab3IP in inducing proliferation of GC cells through its involvement in autophagy. miR-532-3p directly targets Rab3IP and represses its function, thereby demonstrating a novel regulatory link in GC.