〔Abstract〕 Objective To study the impact of high expression level of HK2 gene in human prostate cancer on T help 1/T help 2,which are co-cultured with human prostate cancer(PC3 cell line). Methods shRNA was used to construct a stable transgenic cells(sh-HK2) of PC3 cell line with shRNA-infected PC3 cell line induced by lentivirus infection, nonsense sequences was used to construct lentivirus, PC3 cell line was infected to obtain negative control cells(NC),then the result of knocking down was inspected through qPCR and Western blot.Isolated cancer cells to obtain supernatant after culturing PC3 cells(NC and sh-HK2) for 48 hours. Human peripheral blood mononuclear cells(PBMCs) were extracted from healthy persons. TH1 blank group: PBMCs were cultured in 1640 medium for 48 h under TH1 induction conditions, and TH1 experimental group(TH1 sh-HK2 group): PBMCs were co-cultured for 48 h in TH1 induction conditions using the supernatant of the cultured stable transgenic cells.(TH1 NC group): PBMCs were co-cultured for 48 h under the condition of TH1 induction using the supernatant of the cultured negative control cells. The induction conditions of the TH2 blank group, the TH2 experimental group(TH2 sh-HK2 group), and the TH2 negative control group(TH2 NC group) were changed to TH2 induction conditions, and the others were unchanged. After 48 hours, the lactate level in each co-culture system was detected by a spectrophotometer. PBMCs were then extracted and cultured with phytohemagglutinin-M(PHA-M) for another 24 h, then the secretion of IFN-γ of PBMCs in TH1 induction conditions and IL-4 of PBMCs in TH2 induction conditions were inspected through ELISA. Results qPCR and Western blot showed that HK2 gene knockdown was significant.In TH1 experimental group of TH1-inducing conditions,the secretion of IFN-γ of PBMCs raised. In TH2 experimental group of TH2-inducing conditions,the secretion of IL-4 of PBMCs decreased. The level of lactic acid in sh-HK2 group declined compared with NC group. Conclusion The high expression level of HK2 gene in PC3 cell line enhances lactic acid in tumor environment. At the same time, it decreases the secretion of IFN-γ and enhances the secretion of IL-4, which leads to weak cellular immune function,therefore humoral immunity that invalid for tumor immunity is enhanced, resulting in the progress of tumor.