Effect of PLC-beta 1/CaM signaling pathway mediated by AT1R on the occurrence and development of hepatocellular carcinoma
CANCER CELL INTERNATIONAL(2021)
Abstract
Objective: To study the roles of AT1R, PLC-beta 1, CaM and other related signal molecules in the formation and development of hepatocellular carcinoma (HCC) and their correlation. Methods: ELISA and immunohistochemistry were used to analyze the expressions of target proteins in serum and liver tissue of HCC patients, and the correlation between AT1R, PLC-beta 1 and CaM and postoperative survival status of patients was followed up and determined. CCK-8 method was used to screen the doses of Ang II and candesartan sensitive to HepG2 and HCCLM3 cells. Transwell experiment was used to observe the effects of different drugs on the migration and invasion activity of HCC cells. Meanwhile, flow cytometry and Western blot were used to detect the expression levels of AT1R, PLC-beta 1 and CaM in the cells. Then PLC-beta 1 siRNA was selected to transfect HCC cells, so as to further clarify the mechanism of the above signal proteins. HepG2 cells were inoculated under the hepatic capsule of mice to induce the formation of HCC in situ. Ang II and candesartan were used to stimulate HCC mice to observe the difference in liver appearance and measure the liver index. Finally, ELISA and immunofluorescence experiments were selected to analyze the levels of target proteins in mouse serum and liver tissue. Results: The expression levels of target proteins in serum and liver tissue of HCC patients were significantly increased, and the postoperative survival time of patients with high expression of AT1R, PLC-beta 1 or CaM was obviously shortened. Ang II and candesartan could significantly promote and inhibit the motility of HCC cells, and had different effects on the levels of AT1R, PLC-beta 1 and CaM in cells. However, in hepatocellular carcinoma cells transfected with PLC-beta 1 siRNA, the intervention ability of drugs was obviously weakened. Ang II could significantly promote the formation and progression of mouse HCC, while candesartan had the opposite effect. Meanwhile, medications could affect the expressions of target proteins in mouse serum and liver tissue. Conclusion: AT1R, PLC-beta 1 and CaM may be risk factors affecting the formation and prognosis of HCC, and the PLC-beta 1/CaM signaling pathway mediated by AT1R is an important way to regulate the migration and invasion activity of HCC cells.
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Key words
Hepatocellular carcinoma, AT1R, PLC-beta 1, CaM, Migration, Invasion
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