Objective We conducted an update meta-analysis aiming to verify the association between p27-V109G polymorphism and cancer risk, particular for prostate cancer (PCa). Then, we conducted a case-control study of Han Chinese in central China to verify the evidence-based results. Methods Relevant studies were collected from diverse databases up to March 2017. In addition, a hospital-based (H-B) case-control study enrolling 90 PCa patients and 140 healthy controls was included to verify these evidence-based findings. Genetic risk was calculated by odds ratio (OR) with its corresponding 95% confidence interval (CI). The p27-V109G polymorphism was determined by MassARRAY genotyping method. Results Finally, twenty-four published studies comprising 9627 cases and 12,102 controls were enrolled for the current meta-analysis. Overall analysis suggested that p27-V109G polymorphism decreased overall cancer risk in allelic contrast, heterozygote, and dominant models. When stratified analysis was conducted by ethnicity, data revealed that p27-V109G polymorphism was associated with a decreased cancer risk in Caucasians. Highlighted in the subgroup analysis by cancer type, we uncovered a significantly decreased risk of PCa in allelic contrast, dominant, homogeneous, and recessive models. However, in the validation case-control study, we failed to uncover a positive association between p27-V109G polymorphism and PCa risk. In addition, negative results were also identified when subgroup analyses were stratified by age, tumor grade, tumor stage, PSA levels, and other measurements. Conclusion Although evidence-based results suggest that p27-V109G polymorphism plays a protective role in overall cancer risk, particularly for PCa, our case-control study failed to validate any association between this particular polymorphism and PCa risk.