Abstract: Objective  To investigate the effect of chronic receptor potential 8 (TRPM8) on the invasion and metastasis of hepatocellular carcinoma(HCC) cells and its related regulation mechanism. Methods A total of 20 pairs of matched HCC tissues and their adjacent tissues in our hospital from March 2017 to December 2019 were collected, and the expression of TRPM8 in HCC tissues was detected by immunohistochemistry and qRT-PCR. HCC Huh7 cells were transfected with shTRPM8(shTRPM8 group), and set up a negative control group (shcontrol group). The ability of cell migration and invasion were detected by scratch healing assay and Transwell chamber assay, respectively; the expression of E-cadherin and vimentin were detected by immunofluo-rescence experiment; the expression of AFP, TRPM8 and AKT/GSK-3β/Snail signal pathway related proteins were detected by Western blot. The constructed Huh7-LV-shTRPM8 HCC cells were injected into BALB/C nude mice through the tail vein as the experimental group(LV-shTRPM8 group), while the nude mouse model established by the injection of Huh7-LV-shNC cells were used as the control group(LV-shNC group). The lung metastasis of nude mice was observed after injection for 6 weeks. Results TRPM8 was highly expressed in HCC tissues and cell lines(P<0.01). Compared with the shcontrol group, the migration and invasion of Huh7 cells after TRPM8 silence were decreased (P<0.01);the expression of E-cadherin increased(P<0.001), while the expression of vimentin decreased (P<0.001); the expression of TRPM8 and Snail proteins were down-regulated(P<0.001), and the phosphorylation degree of AKT and GSK-3β proteins were decreased (P<0.001). TRPM8 silencing decreased the lung metastasis rate, the number of lung metastatic nodules and the expression of AFP and TRPM8 in lung tissues in nude mice (P<0.01). Conclusion TRPM8 can inhibit the invasion and metastasis of HCC Huh7 cells by regulating the AKT/GSK-3β/Snail signal pathway.

Key words: Hepatocellular carcinoma, TRPM8, AKT/GSK-3β/Snail signaling pathway, Invasion, Metastasis