Wu Zhiyong, Kong Xiaoyu, Wang Zhihui
Department of General Surgery, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430014, P.R. China.
Exp Ther Med. 2021 Apr;21(4):309. doi: 10.3892/etm.2021.9740. Epub 2021 Feb 1.
The present study aimed to investigate the effects of integrin α7 (ITGA7) on regulating hepatocellular carcinoma (HCC) progression and endothelial-mesenchymal transition (EMT). ITGA7 mRNA and protein expression in human normal liver epithelial cells and HCC cell lines were determined by reverse transcription-quantitative PCR (RT-qPCR) and western blotting. ITGA7 small interfering RNA [siRNA; ITGA7-knockdown (KD) group] and nonsense siRNA (control group) were transfected into Huh7 cells and SNU449 cells, respectively. ITGA7 mRNA and protein expression (RT-qPCR and western blotting, respectively), cell proliferation (Cell Counting Kit-8 assay), apoptosis (annexin V/propidium iodide assay), migration (wound scratch assay) and invasion (Transwell assay) were then detetected. E-cadherin, α-smooth muscle actin (α-SMA), vimentin and V-cadherin levels (RT-qPCR and western blotting) were also assessed. ITGA7 mRNA and protein expression levels were increased in Li7, Huh7, SKHEP1 and SNU449 cells compared with THLE-3 cells. Following transfection, ITGA7 mRNA and protein expression was lower in the ITGA7-KD group compared with that in the control group in both Huh7 and SNU449 cells, indicating successful transfection. In the ITGA7-KD group, cell proliferation decreased at 48 and 72 h, cell apoptosis rates increased at 48 h, cell migration rate was reduced at 24 h and cell invasion decreased at 24 h compared with the control group. Additionally, increased E-cadherin but decreased α-SMA, vimentin and V-cadherin mRNA and protein expression levels were observed in the ITGA7-KD group compared with the control group at 24 h. In conclusion, ITGA7 knockdown suppressed HCC progression and inhibited EMT in HCC , implying that ITGA7 might be a novel treatment target for HCC.
本研究旨在探讨整合素α7（ITGA7）对调节肝细胞癌（HCC）进展及内皮-间充质转化（EMT）的影响。通过逆转录定量聚合酶链反应（RT-qPCR）和蛋白质印迹法检测人正常肝上皮细胞及HCC细胞系中ITGA7 mRNA和蛋白表达。分别将ITGA7小干扰RNA [siRNA；ITGA7基因敲低（KD）组]和无义siRNA（对照组）转染至Huh7细胞和SNU449细胞。随后检测ITGA7 mRNA和蛋白表达（分别采用RT-qPCR和蛋白质印迹法）、细胞增殖（细胞计数试剂盒-8法）、凋亡（膜联蛋白V/碘化丙啶法）、迁移（划痕实验）和侵袭（Transwell实验）。还评估了E-钙黏蛋白、α-平滑肌肌动蛋白（α-SMA）、波形蛋白和V-钙黏蛋白水平（RT-qPCR和蛋白质印迹法）。与THLE-3细胞相比，Li7、Huh7、SKHEP1和SNU449细胞中ITGA7 mRNA和蛋白表达水平升高。转染后，Huh7和SNU449细胞中ITGA7-KD组的ITGA7 mRNA和蛋白表达均低于对照组，表明转染成功。与对照组相比，ITGA7-KD组在48小时和72小时时细胞增殖减少，48小时时细胞凋亡率增加，24小时时细胞迁移率降低，24小时时细胞侵袭减少。此外，与对照组相比，ITGA7-KD组在24小时时E-钙黏蛋白增加，但α-SMA、波形蛋白和V-钙黏蛋白的mRNA和蛋白表达水平降低。总之，ITGA7基因敲低可抑制HCC进展并抑制HCC中的EMT，这意味着ITGA7可能是HCC的一个新治疗靶点。
