The purpose of the present study is to detect the association between the levels of testosterone with prostate cancer patients by the evaluation of a tumor marker Prostate-specific antigen (PSA). Prevalence was recorded in Gujrat, Punjab, Pakistan. Blood samples of normal and prostate cancer males were taken from Gujrat, Punjab, Pakistan. Information including age, marital status, smoking, Prostate-specific antigen (PSA), testosterone, and any previous record was obtained with the patient’s consent. Exclusion and inclusion criteria were defined. The samples of control and patients were analyzed through ELISA protocol to check the testosterone and Prostate-specific antigen (PSA). Testosterone significantly affects the prostate-specific antigen (PSA) with (P<0.05). In the cases, the mean testosterone, age, PSA was 38.45±36.66, 66.28±6.386 and 9.655±6.656, respectively while in the control group mean testosterone, age and PSA were 194.1±84.08, 59.35±6.794 and 1.295±0.809 respectively. The present study concludes testosterone levels as negatively affected by prostate specific antigen (PSA). The prevalence of Prostate cancer recorded in Gujrat, Punjab, Pakistan was 2%. Our study may prove feasible for the detection of prostate cancer in aged males. Testosterone replacement therapy may prove to be effective in retrieving the complications induced by prostate cancer.