The role of GPRC6A in prostate cancer cell on proliferation and apoptosis / 医学研究生学报

ABSTRACT

Objective　The incidence of prostate cancer (PCa) is increasing year by year in China. G protein coupled receptor family C,group 6,member A (GPRC6A ) is a susceptible gene of PCa found in recent years. To investigate therole of GPRC6A and extracellular signal-regulated kinase (ERK) mediated by GPRC6A in the proliferation and apoptosis of PCaLncapC4-2 cells, and provide new ideas for targeted therapy of PCa.　Methods　The experiment was divided into the LncapC4-2 PCDH group (only the empty vector pCDH was transfected), the LncapC4-2 GPRC6A group (only the pCDH-GPRC6A was transfected) and the LncapC4-2 GPRC6A + U0126 group (pCDH-GPRC6A was transfected and treated with U0126). The proliferation of LncapC4-2 cells was detected by using CCK8 kit. The changes of cell cycle and apoptosis were detected using flow cytometry. Cell cycle and apoptosis-related proteins were evaluated by Western blot.　Results　The result ofWestern blots showed the expression ofGPRC6A and P-ERK in LncapC4-2 GPRC6A group washigher than that in LncapC4-2 PCDH group(P<0.05), and the expression of P-ERKin LncapC4-2 GPRC6A + U0126 group was evidently lower than that in LncapC4-2 GPRC6A group(P<0.05). The result of CCK8 tests showed the cell proliferation ability in LncapC4-2 GPRC6A group was higher than that in LncapC4-2 PCDH group, and the proliferation ability in LncapC4-2 GPRC6A + U0126 group was lower than that in LncapC4-2 GPRC6A group, and there was statistical difference between two groups(P<0.05).Flow cytometry results showed the ratio of G1/（G2+S）in LncapC4-2 GPRC6A groupwas evidently lower than that in LncapC4-2 PCDH group(P<0.05), and the ratio of G1/(G2+S)in LncapC4-2 GPRC6A + U0126 group was evidently higher than that in LncapC4-2 GPRC6A group(P<0.05). Western blot results showed that the expression of CyclinD1 were evidently increased in LncapC4-2 GPRC6A group(0.88±0.04) compared to LncapC4-2 PCDH group(0.66±0.02, P<0.05), the expression CyclinD1 wasevidently reduced in LncapC4-2 GPRC6A +U0126 group(0.71±0.02) compared with LncapC4-2 GPRC6A group (P<0.05). The ratio of apoptosis in LncapC4-2 GPRC6Agroup(3.64%) was evidently reduced than that in LncapC4-2 PCDH group(9.00%) and in LncapC4-2 GPRC6A +U0126 group(19.57%, P<0.05). Meanwhile, western blot results showed that the expression of BCL2 was increased and the expression of Cleaved-caspase3 was lower in LncapC4-2 GPRC6A group compared to LncapC4-2 PCDH group (P<0.05). The expression of BCL2 was reduced and the expression of Cleaved-caspase3 was increased in LncapC4-2 GPRC6A +U0126 group compared with LncapC4-2 GPRC6A group (P<0.05).　Conclusion　GPRC6A may be promote the proliferation and inhibit apoptosis of PCa cells through ERK signaling pathway, thus to be involved in the development of PCa .