Overexpression of pressure-responsive miRNA-5703 inhibits pressure-induced growth and metastasis of liver cancer

Si Shen; Wenli Zhou; Ji Xuan; Weijun Xu; Huabing Xu; Miaofang Yang; Liang Zhu; Zhuoxin Yang; Benzhao Yang; Bin Shi; Ying Zhao; Fangyu Wang

doi:10.7150/jca.64926

Overexpression of pressure-responsive miRNA-5703 inhibits pressure-induced growth and metastasis of liver cancer

J Cancer. 2022 Jan 1;13(1):325-342. doi: 10.7150/jca.64926. eCollection 2022.

Authors

Si Shen^{1

2}, Wenli Zhou³, Ji Xuan¹, Weijun Xu¹, Huabing Xu¹, Miaofang Yang¹, Liang Zhu², Zhuoxin Yang¹, Benzhao Yang⁴, Bin Shi², Ying Zhao⁵, Fangyu Wang¹

Affiliations

¹ Jinling Hosp Dept of Gastroenterology and Hepatology, Nanjing Univ, Sch Med, Nanjing 210002, P R China.
² Changzheng Hosp Dept of Gastroenterology, Naval Med Univ, Shanghai 200003, P R China.
³ Changzheng Hosp Dept of Oncology, Naval Med Univ, Shanghai 200003, P R China.
⁴ Dept of Cardiology, Naval Medical Center, Naval Med Univ, Shanghai 200005, P R China.
⁵ Changzheng Hosp Dept of Traditional Chinese Medicine, Naval Med Univ, Shanghai 200003, P R China.

Abstract

A vast majority of liver cancers coexist with cirrhosis and/or portal hypertension. A high-pressure tumour microenvironment may lead to malignant progression of liver cancer. Through quantitative reverse transcription-polymerase chain reaction, we found that miRNA-5703 was expressed at low levels in HepG2 and Huh-7 cells and pressure-treated MHCC97H implanted mouse cancer tissues, while its potential target gene, sarcoma gene (SRC), was highly expressed. The expression of miRNA-5703 was higher in liver cancer tissues from Barcelona Clinic Liver Cancer (BCLC) stage A1 patients than those from BCLC stage A2-D patients, whereas SRC showed the opposite expression pattern. Bioinformatics analysis, luciferase reporter assay, and western blotting were performed to verify the relationship between miRNA-5703 and its potential target SRC. Using intravital imaging and immunohistochemistry, we demonstrated that pressure promotes tumour growth in subcutaneous tumourigenesis nude mice, and overexpression of miRNA-5703 significantly downregulated Ki67 and upregulated NM23 in tumour tissues of mice, implying the blockage of tumour growth and metastasis. The activation of proliferation, migration, and invasion of HepG2 and Huh-7 cells by pressure, and inhibition by overexpressing miRNA-5703 were observed by cell counting kit-8 assay, flow cycle assay, transwell assay, and wound healing assay. After the intervention of pressure, inhibitor, and lentivirus to hepatoma cells, SRC, focal adhesion kinase (FAK), phosphatidylinositol 3-kinase (PI3K), serum/glucocorticoid regulated kinase-3 (SGK3), phosphoinositide dependent protein kinase 1 (PDK1), and paxillin were upregulated, and forkhead box O1 (FOXO1) and cyclin dependent kinase inhibitor 1B (P27^Kip1) were downregulated in pressure-loaded hepatoma cells, which could be reversed by overexpression of miRNA-5703 or SRC knockdown. In conclusion, upregulation of miRNA-5703 inhibited pressure-induced growth and metastasis by suppressing the SRC-FAK-FOXO1 axis and SRC-paxillin axis. This novel perspective may be conducive to the mechano-inspired anticancer drugs of liver cancer.

Keywords: liver cancer; metastasis; miRNA-5703; portal hypertension; pressure; proliferation.