〔Abstract〕 Objective To explore the differential expression of TM6 SF2 in hepatocellular carcinoma and its effect on the biological behavior of hepatocellular carcinoma cells. Methods Immunohistochemistry was used to detect TM6 SF2 expression in 15 cases of hepatocellular carcinoma and adjacent tissues. Western blot was used to detect the expression of TM6 SF2 in hepatocellular carcinoma cell line Hep G2 and normal liver cell line L-02. TM6 SF2 expression and survival curve in hepatocellular carcinoma were analyzed based on TCGA database. A TM6 SF2 overexpression vector was constructed based on the PCDNA 3.1 plasmid. Hep G2 was infected by lentiviral packaging, and stable cell lines were screened. Western blot was used to detect the TM6 SF2 over-expression efficiency. For the blank group, control group and TM6 SF2 overexpression group, CCK-8 and cell cloning method were used to detect cell proliferation activity, Annexin V-FITC/PI double labeling was used to detect cell apoptosis, and Transwell was used to detect cell invasion ability. Based on the clinical samples of hepatocellular carcinoma tissues, q-PCR was used to detect the expression levels of TM6 SF2 and SLC27 A5, and the expression correlation was analyzed. Results TM6 SF2 expression was significantly reduced in hepatocellular carcinoma tissues and cells. The 5-year survival of hepatocellular carcinoma patients with TM6 SF2 low expression was shorter. After successful construction of TM6 SF2 overexpressing stable cell line Hep G2, TM6 SF2 overexpression could significantly reduce Hep G2 cell proliferation activity, invasion ability, and promote apoptosis. There was a positive correlation between TM6 SF2 and SLC27 A5 expression in hepatocellular carcinoma tissues. Conclution TM6 SF2 shows strong potential of tumor suppressor genes and has certain clinical significance. It provides new ideas and theoretical basis for future clinical diagnosis, prognosis judgment and molecular targeted therapy of hepatocellular carcinoma based on TM6 SF2.