Department of Hepato-Pancreato-Biliary Surgery, Zhongnan Hospital of Wuhan University, Wuhan, China.
College of Life Science, Wuhan University, Wuhan, China.
J Cell Mol Med. 2018 May;22(5):2569-2579. doi: 10.1111/jcmm.13499. Epub 2018 Mar 8.
This study aimed to explore the effects of CLIC1 gene silencing on proliferation, migration, invasion and apoptosis of human gallbladder cancer (GBC). GBC and normal gallbladder tissues were extracted for the detection of mRNA and protein expressions of CLIC1. GBC-SD and NOZ cells in the logarithmic growth phase were selected to conduct the experiment. Three different siRNA recombined expression vectors were established using CLIC1 as a target at different sites. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blotting were, respectively, used to detect the CLIC1 mRNA and protein expressions. MTT assay was performed to detect the cell proliferation. Flow cytometry was applied to measure the cell apoptosis and cell cycle distribution. The variations of cell migration and invasion were evaluated using Transwell assay. GBC tissues showed higher CLIC1 mRNA and protein expressions than normal gallbladder tissues. The CLIC1 mRNA and protein expressions in the CLIC1 siRNA group were significantly lower than those in the NC and blank groups. Compared with the NC and blank groups, the CLIC1 siRNA group showed a significant decrease in cell proliferation but an obvious increase in apoptosis rate in GBC cells. Besides, in the CLIC1 siRNA group, cell percentage in G0/G1 and G2/M phase was gradually increased but decreased in S phases. The migration and invasion abilities in GBC cells were significantly lower than those in the NC and blank groups. Our study demonstrates that CLIC1 gene silencing could promote apoptosis and inhibit proliferation migration and invasion of GBC cells.
本研究旨在探讨 CLIC1 基因沉默对人胆囊癌（GBC）增殖、迁移、侵袭和凋亡的影响。提取 GBC 和正常胆囊组织，检测 CLIC1 的 mRNA 和蛋白表达。选择对数生长期的 GBC-SD 和 NOZ 细胞进行实验。以 CLIC1 为靶点，在不同部位构建 3 种不同的 siRNA 重组表达载体。分别采用逆转录定量聚合酶链反应（RT-qPCR）和 Western blot 检测 CLIC1 mRNA 和蛋白表达。MTT 法检测细胞增殖。流式细胞术检测细胞凋亡和细胞周期分布。Transwell 法检测细胞迁移和侵袭的变化。GBC 组织中 CLIC1 mRNA 和蛋白表达均高于正常胆囊组织。CLIC1 siRNA 组的 CLIC1 mRNA 和蛋白表达明显低于 NC 组和空白组。与 NC 组和空白组相比，CLIC1 siRNA 组 GBC 细胞增殖明显减少，凋亡率明显升高。此外，在 CLIC1 siRNA 组中，G0/G1 和 G2/M 期细胞百分比逐渐增加，而 S 期细胞百分比减少。GBC 细胞的迁移和侵袭能力明显低于 NC 组和空白组。本研究表明，CLIC1 基因沉默可促进 GBC 细胞凋亡，抑制增殖、迁移和侵袭。
