Experimental Study
Expression of Six homeobox 4 protein in hepatocellular carcinoma and its effect on malignant biological behavior
Zhou Yancai, Zhou Bing, Wei Xiaoxia, Wu Hongjie, Sun Lihua, Zhu Bin
Published 2021-02-08
Cite as Chin J Exp Surg, 2021, 38(2): 277-280. DOI: 10.3760/cma.j.cn421213-20200609-01197
Abstract
ObjectiveTo investigate the expression of Six homeobox 4(Six4) protein in hepatocellular carcinoma (HCC) and its effects on the proliferation, migration and epithelial mesenchymal transition of HCC cells.
MethodsA total of 169 cases of HCC tissues and corresponding adjacent tissues from June 2018 to June 2019 were selected as the research objects, and the expression levels of Six4 protein in liver cancer tissues and adjacent tissues were analyzed by immunohistochemistry. Six4 knockout cell lines were established in HepG2 cells by clustered regularly interspaced short palindromic repeats-cas9 (CRISPR-Cas9), named as control group and Six4 KO group. The cell proliferation in the control group and Six4 KO group was analyzed by cell counting kit-8 (CCK-8) and 5-ethynyl-2′-deoxyuridine (EdU) staining. The cell migration in the two groups was analyzed by Transwell. The expression of N-cadherin, E-cadherin and vimentin was analyzed by Western blotting.
ResultsCompared with the adjacent tissues (79.43±10.43), the expression level of Six4 protein in HCC tissues (169.59±18.59) was significantly increased (t=4.179, P<0.05). Six4 knockout cell line was successfully constructed by CRISPR-Cas9. Compared with the control group (2.15±0.28), the cell absorbance of Six4 KO group (1.21±0.19) significantly decreased (t=3.441, P<0.05). The positive rate of EdU staining in the control group (76.45±8.90)% was significantly higher than that in the Six4 KO group (31.63±7.43)%, and the difference was statistically significant (t=4.693, P<0.05). Compared with the control group (119.48±12.08), the number of migration cells in the Six4 KO group (65.64±9.99) significantly decreased (t=3.748, P<0.05). Compared with the control group (0.33±0.11), the relative expression level of E-cadherin in the Six4 KO group was significantly increased (1.49±0.21), and the difference was statistically significant (t=5.184, P<0.05). Compared with the relative expression levels of N-cadherin and vimentin in the control group (1.74±0.28, 1.89±0.26), the relative expression levels of N-cadherin and vimentin in the Six4 KO group (0.58±0.11, 0.68±0.23) were significantly decreased (t=4.274, P<0.05;t=4.109, P<0.05).
ConclusionSix4 protein is highly expressed in HCC tissues, which is involved in the proliferation, migration and epithelial mesenchymal transition of HCC cells.
Key words:
Six homeobox 4; Hepatocellular carcinoma; Proliferation; Migration; Epithelial mesenchymal transition
Contributor Information
Zhou Yancai
Department of Infectious Diseases, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang 453100, China
Zhou Bing
Department of Hepatobiliary and Pancreatic Surgery, the First Affiliated Hospital of Xinxiang Medical College, Xinxiang 453100, China
Wei Xiaoxia
Department of Infectious Diseases, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang 453100, China
Wu Hongjie
Department of Infectious Diseases, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang 453100, China
Sun Lihua
Department of Infectious Diseases, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang 453100, China
Zhu Bin
Department of Infectious Diseases, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang 453100, China
