The Effect Of E2F3in The Progression Of Castration Resistant Prostate Cancer

Objective:To detect the expression of E2F3protein between androgen dependent prostate cancer cell line(LNCAP) and androgen independent prostate cancer cell line (C4-2,PC-3,DU145), to detect the role of E2F3in the progression of prostate cancer to androgen independence and explore the mechanism of the progression of casrtation resistant prostate cancer.Methods:The expression of E2F3protein in prostate cancer cell lines (LNCAP,C4-2,PC-3,DU145) was detected by westernblot;The plasmid pcDNA3.1-HA-E2F3-GFP was used to enforce overexpression of E2F3in LNCAP cell.The stable overexpression cells were screened out,named LNCAP-E2F3.LNCAP cell line and LNCAP-E2F3cell line were maintained in RPMI1640supplemented with10%fetal calf serum and with10%Charcoal-stripped FBS.96hours later.the cell survival and proliferation was determined by MTT study.Cell apoptosis index was detected by flowcytometry.Result:1.The expression of E2F3protein are various in prostate cancer cell lines. The relative gray scale values of LNCAP, C4-2,PC-3and DU145cells were1.1095±0.0113,2.0492±0.0192,2.4395±0.0854,2.7095±0.0501,p<0.05,the differences were statistically significant.2.The E2F3protein of LNCAP cell were significantly increased after transfected by pcDNA3.1-HA-E2F3-GFP.We successful constructed LNCAP-E2F3cell line,in which the expression of E2F3protein are sigificantly higher compared to LNCAP cell line.3.The rate of survial of LNCAP-E2F3,LNCAP and empty plasmid group cells were0.804±0.013,0.807±0.0150,1.593±0.031when maintained in RPMI1640supplemented with10%Charcoal-stripped FBS. The rate of survial of LNCAP-E2F3cell line are higher than LNCAP cell line when both are maintained in RPMI1640supplemented with10%Charcoal-stripped FBS.4. The apoptosis index of LNCAP-E2F3,LNCAP and empty plasmid group cells were (24.47±1.10)%,(47.46±0.60)%,(45.56±0.61)%when maintained in RPMI1640supplemented with10%Charcoal-stripped FBS after72hours.Compared to the others,the apotosis index of LNCAP-E2F3cell lines was significantly decreased.Conclusion:E2F3is highly expressed in androgen independent prostate cancer cell line.E2F3play a critical role in the progression of castration resistant prostate cancer.The result suggest that E2F3would be a biomarker candidates for prostate cancer.