Study On The Role Of GPRC6A In Epithelial-Mesenchymal Transition Of Prostate Cancer Cells

Objective:In recent years,prostate cancer(PCa)is becoming one of the malignant tumor of urinary system which seriously affects the male health of our country.In PCa patients,bone is the most common site of metastasis.Bone metastasis is also the main cause of PCa death.It is necessary to find the markers of PCa metastasis and to provide new ideas and new targets for the effective prevention and control of PCa metastasis.Studies have shown that epithelial-mesenchymal transiton(EMT)is one of the most important phenotypes of tumour metastasis and invasion.G protein-coupled receptor family C group 6 member A(GPRC6A)is widely expressed in many human tissues and organs,and plays an important role in the regulation of energy metabolism and sex hormone.GPRC6 A is highly expressed in PCa cells and can promote the progress of PCa,but the specific mechanism is not clear.This research intends to study the cell and animal level,to clarify the role of GPRC6 A in the PCa and explore the regulation mechanism in process of EMT on PCa cells to further improve the regulatory mechanism of EMT in PCa,provide a new way to explore molecular targets for clinical gene treatment of PCa.Methods:The lentiviral vector with GPRC6 A over expression was constructed to infect LNcapC4-2 cells,then screen of stable expression clones;using the constructed cell lines as the research object,By using QPCR,Western blot,scratches and Transwell.The effect of GPRC6 A on the EMT related phenotypes and metastasis ability of PCa cells was detected preliminarily;Subcutaneous tumor formation experiment,immunohistochemistry and Western blot are used to confirm the influence of GPRC6 A in processof EMT of PCain vivo.followed by TGF?1 stimulated,by using Western blot and immunofluorescence to detect the expression of GPRC6 A and EMT respectively,while using transwell to assay TGF?1 induced cell metastasis mediated by GPRC6 A,to Study on the role of GPRC6 A in epithelial-mesenchymal transition of prostate cancer cells Results:The GPRC6 A overexpressed LNcapC4-2 cell line of PCa was constructed successfully.In vitro,The overexpression of GPRC6 A could increase the transfer ability of LNcapC4-2 in PCa cells,and also change the EMT related phenotypes;in vivo,GPRC6 A can promote tumor formation with changes in the expression of EMT related factors;After induction of TGF?1,it does not cause obvious cell morphologic changes,but changes the expression of EMT related factors and promotes cell metastasis.GPRC6 A can exacerbate this change.Conclusion:GPRC6A may be a new PCa metastasis promoting factor that can promote the EMT process in PCa cells and then affect the development of PCa.