Infection with specific high-risk human papillomavirus (HPV) types 16 and 18 have been strongly associated with the genesis of various neoplasms in humans, though such study in lung cancer is limited and the results are controversial. In the present study, we collected and explored 313 fresh lung tumor specimens for the presence of HPV with polymerase chain reaction and non-isotopic in situ hybridization. We found that 44.1% of (138/313) non-small cell lung carcinoma (NSCLC) samples were positive for HPV detection, while 4.2% (4/96) of lung benign controls were positive for HPV 16 and 18 DNA. HPV infection was significant between lung squamous cell carcinoma and adenocarcinoma as well as smoking and non-smoking patients. In HPV-positive lung cancer tissues, abnormal p53 protein accumulation was seen in 97 of the 138 carcinomas (70.3%) and expression of pRb in 54 of the 138 carcinomas (39.1%). There was an obvious relationship between the presence of papilloma viral DNA and abnormal p53 protein accumulation and pRb depletion. Cell proliferation and apoptosis were correlated with HPV infection in NSCLC samples. Our data confirm the high prevalence of HPV in lung carcinomas in the central part of China and suggest the possible mechanism of the carcinogenic role of HPV in these carcinomas.