Objective To discuss the correlation of the p53 overexpression and human papilloma virus (HPV) infection with the occurrence, development of the vulvar squamous cell carcinoma in elderly women. Methods Samples from 68 patients with vulvar squamous cell carcinoma and samples of formalin-fixed paraffin-embedded blocks from 38 patients with vulvar dystrophy were determined by immunohistochemistry. HPV infection was examined with monoclonal or polyclonal antibodies. Fifteen normal vulvar skin samples were used as controls. Results Positive expression rate of p53 and HPV in vulvar squamous cell carcinoma samples were 54.4% and 45.6% respectively, in the dystrophy samples were 18.4% and 55.3% respectively. All showed significant difference between the two groups. None was positive in the controls. Conclusions There are high expression of p53 and HPV in the elderly women with vulvar squamous cell carcinoma. p53 mutation and high risk HPV infection play an important role in the development of vulvar squamous cell carcinoma.