The invention belongs to the technical field of non-coding nucleic acids for regulating gene expression, and discloses a microRNA molecule related to 5-fluorouracil resistance and application thereof.The microRNA molecule is MiR-195, and an MiR-195 nucleotide sequence is SEQ ID NO:1. The microRNA molecule which may be involved in hepatocellular carcinoma cell line 5-fluorouracil resistance is predicted by a bioinformatics technology, a lentiviral vector is constructed based on a molecular cloning related technology, clear biological function changes of liver cancer cells infected with microRNA virus are utilized to screen out the microRNA clearly participating in liver cancer cell 5-fluorouracil-resistance, a biological function and mechanism of the screened miR-195 promoting the of livercancer cell drug resistance are further analyzed and verified, and an effective path is provided for effective reversal of liver cancer 5-fluorouracil resistance and improvement of a clinical chemotherapy effect on liver cancer.