Objective To explore the effect of transcription factor B1,mitochondrial,(TFB1M) on cell survival and migration of hepatocellular carcinoma cells and related molecular mechanisms. Methods TFB1M gene was overexpressed by transfection of eukaryotic recombinant plasmid. Cell survival and cell cycle distribution were detected by CCK-8 and flow cytometry. Cell migration and invasion were assessed by scratch healing and transwell chamber assays. Results The expression of TFB1M was significantly decreased in HCC, and the expression of TFB1M was more lower in female HCC patients than that tin male HCC patients. The mRNA expression of TFB1M was significantly increased after transfection of recombinant plasmid. Moreover, our results showed that overexpression of TFB1M had no effect on the growth and cell cycle distribution of HepG2 and Huh7 cells. However, overexpession of TFB1M remarkably attenuated the migration and invasion activity of HCC cells(all P<0.05). The results from the bioinformatics presented that the low expression of TFB1M might be related to the extracellular matrix assembly gene set in HCC. Conclusions TFB1M regulates the cell metastasis and invasion through influencing extracellular matrix assembly in HCC.