Objective This paper is to investigate the effects and mechanisms of methylenetetrahydrofolate dehydrogenase 2(MTHFD2)on the proliferation,migration,and invasion of liver cancer cells.Methods A total of 58 patients with hepatocellular carcinoma(HCC)who underwent liver cancer resection at the First People's Hospital of Fuyang of Hangzhou from April 2020 to August 2023 were selected as the study subjects.The surgically resected HCC tissues and adjacent tissues were collected,and the expression level of MTHFD2 in the tissues was detected by immunohistochemical staining.Normal liver cell line LO2,as well as human liver cancer cell lines HepG2,Hep3B,and SMMC-7721 were selected,and real-time fluorescence quantitative PCR and Western blotting were used to detect the expression level of MTHFD2 in each group of cells.The HepG2 cells were divided into the blank control group(given buffer solution only),the si-NC group(transfected with si-NC),and the si-MTHFD2 group(transfected with si-MTHFD2).In addition,the proliferation,migration,and invasion abilities of each group of cells were compared.Thirty BALB/C-nu/nu male nude mice were selected,and the HepG2 cells in logarithmic growth phase after transfection for 24 hours were implanted in the right axilla of the mice.The tumor volume was measured weekly,and after 4 weeks,the tumor was removed and weighed.Results The positive expression rate of MTHFD2 protein in the HCC tissue is 70.69%higher than that in adjacent tissues(32.76%),with a statistically significant difference(P＜0.05).Compared with LO2 cells,the protein and mRNA expression levels of MTHFD2 are significantly increased in HepG2,Hep3B,and SMMC-7721 cells(P＜0.05),which are the highest in HepG2 cells.Compared with the blank control group and the si-NC group,the protein and mRNA expression levels of MTHFD2 in the si-MTHFD2 group are significantly reduced(P＜0.05).After transfection for 72 and 96 hours,the OD450 values of the si-MTHFD2 group are significantly lower than those of the blank control group and the si-NC group(P＜0.05).Compared with the blank control group and the si-NC group,the si-MTHFD2 group shows a decrease in cell scratch healing rate,a reduction in the number of invasive cells,and a significant decrease in the expression levels of phosphoinositol 3-kinase(PI3K),phosphorylated PI3K(p-PI3K),protein kinase B(AKT),p-AKT,mammalian target of rapamycin(mTOR),and p-mTOR protein in the cells(P＜0.05).After 3 weeks of cultivation,the tumor volume in the si-MTHFD2 group of nude mice is smaller than that in the blank control group and the si-NC group(P＜0.05).After 4 weeks of cultivation,the weighing results show that the tumor mass in the si-MTHFD2 group of nude mice is significantly smaller than that in the blank control group and the si-NC group(P＜0.05).Conclusions Downregulation of MTHFD2 expression can inhibit the proliferation,migration,invasion,and in vivo tumorigenicity of liver cancer cells,whose mechanism may be related to the negative regulation of PI3K/AKT/mTOR signaling pathway transduction.MTHFD2 is expected to become a new target for the treatment of HCC.
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