The invention discloses shRNA for inhibiting human EDARADD gene expression, a lentiviral vector and a construction method and application of the lentiviral vector. The method comprises the following steps: firstly, synthesizing a positive strand and a negative strand of DNA oligo aiming at an RNA interference target sequence; annealing the positive strand and the negative strand to form double-stranded DNA with a cohesive end; linearizing the lentiviral vector through enzyme digestion, and connecting the linearized vector with double-stranded DNA; transforming escherichia coli competent cellswith a connection product, and carrying out PCR identification on the obtained positive clone; and performing plasmids extraction for virus packaging in the next step. According to the invention, theRNA interference target sequence is designed by taking an EDARADD gene as a template, an shRNA interference sequence is designed according to a selected target sequence, an RNA interference lentiviralvector is constructed through packaging, and after the EDARADD gene is knocked down, proliferation of tongue squamous cell carcinoma cells CAL-27 can be remarkably influenced, the cell clone formingcapacity is influenced, and cell apoptosis is promoted.