The Fuction And Mechanism Study Of Histamine H4 Receptor In Prostate Cancer

Part Ⅰ Expression of H4 R in prostate cancer Objective: To explore the different expression of H4 R in prostatic hyperplasia tissue and prostate cancer Methods: 10 cases of prostate cancer and 10 cases of prostate tissue were collected from guangxi medical university affiliated tumor hospital surgical specimen archive wax block between April 2014 and April 2015.Immunohistochemistry and Real time PCR were performed to test the expression of H4 R in 10 cases of prostate cancer and 10 cases of prostatic hyperplasia Results: Compared with prostatic hyperplasia,the expression of H4 R was increased obviously in prostate cancer(P<0.05).Conclusions: H4 R is mainly expressed in prostate cancer.Part Ⅱ The study of H4 R on invasion of prostate cancer cells PC3 and LNcap Objective: To explore the effects of H4 R on invasion of prostate cancer cells Methods: The prostate cancer cells PC3 and LNcap were used as the research object.(1)Real time PCR and Immunofluorescence staining were carried out to detect the expression of H4 R in the two kinds of cell lines.(2)The above two kinds of cells respectively disposed by H4 R agonist 4-methylhistamine and4-methylhistamine inhibitor JNJ7777120 were divided into four groups: control group,4-methylhistamine group,JNJ7777120 group,and 4-methylhistamine +JNJ7777120 group.The expression of H4 R in the four group cells were tested by Real time PCR.Real time PCR and western blot were carried out to detect the expression of m RNA and protein of MMP2 and MMP9 in control group,4-methylhistamine group,and 4-methylhistamine + JNJ7777120 group.Results: The expression of H4 R in PC3 cell line was significantly higher than in LNcap cell line(P<0.05).Disposed by 4-methylhistamine,the expression of H4 R in two types cells was increased(P<0.05),while,compared with control group,the expression of H4 R in JNJ7777120 group,and 4-methylhistamine +JNJ7777120 group had no significantly difference(P>0.05).The expression of m RNA and protein of MMP2 and MMP9 in PC3 cell line were significantly higher than in LNcap cell line(P<0.05).Conclusions: H4 R can promote the invasion and metastasis of prostate cancer cellsPart Ⅲ The mechanisms underlying the invasion induced by H4 R in human prostate cancer cells Objective: To explore the mechanisms of invasion induced by H4 R in prostate cancer cells Methods: The prostate cancer cell LNcap was used as the research object,(1)The cell LNcap was transfected with H4 R Si RNA and gained thestable interference model namely H4 R Si RNA group.Real time PCR and western blot were carried out to detect the expression of H4 R in H4 R Si RNA group.(2)Disposed by 4-methylhistamine and JNJ7777120 to gain 4-methylhistamine group and JNJ7777120 group.Real time PCR,Immunofluorescence staining and Immunohistochemistry were carried out to detect the expression of TGF-β1,GD3 S,E-cadherin and Vimentin between control group,4-methylhistamine group and JNJ7777120 group.Results :(1)Experiments in prostate cancer cell line stably-transfected with H4 R Si RNA.The expression of H4 R in H4 R silencing cell was decreased obviously compared with control group(P<0.05).(2)During control group、H4R Si RNA group、4-methylhistamine group and JNJ7777120 group.In the 4-methylhistamine group,expression of TGF-β1 、GD3S and Vimentin were higher than control group,while expression of E-cadherin was decreased(P<0.05).In H4 R Si RNA group,expression of GF-β1、GD3S and Vimentin were lower than control group,while expression of E-cadherin was increased(P<0.05).Conclusions:(1)The cancer cell transfected H4 R by Si RNA and stably expressed interference effects in LNcap cell.H4 R interference inhibit prostatecancer cell invation.(2)Increasing the expression of TGF-β1,GD3 S and Vimentin indicate that H4 R activation can promote the process of EMT.Decreasing the expression of TGF-β1,GD3 S and Vimentin indicate that H4 R inhibition can suppress the process of EMT.