目的：构建含TFDP3基因启动子和荧光素酶报告基因的重组载体pGL3-TFDP3-promoter，观察E2F1对TFDP3转录及表达的调控作用以及TFDP3对E2F1诱导肿瘤细胞凋亡的影响。方法：以人前列腺癌PC3细胞系基因组DNA为模板，PCR扩增TFDP3启动子序列并克隆入荧光素酶报告基因载体，与E2F1表达载体pCMV-E2F1-HA瞬时单独或共同转染PC3细胞，测定荧光素酶活性以观察E2F1对TFDP3启动子的调控作用，Western blotting检测pCMV-E2F1-HA转染对PC3细胞内TFDP3表达的影响，流式细胞术检测TFDP3与E2F1相互作用对前列腺癌细胞凋亡的影响。结果：成功构建TFDP3基因启动子重组质粒pGL3-TFDP3-promoter，与E2F1表达载体pCMV-E2F1-HA共转染PC3细胞后，TFDP3启动子诱导的荧光素酶活性较单独转染pGL3-TFDP3-promoter显著升高[（1.14±0.06）vs（0.61±0.05）, P<0.05]。转染pCMV-E2F1-HA的PC3细胞的TFDP3蛋白表达是未转染细胞的2.7倍[（0.24±0.03）vs（0.09±0.02）, P<0.05]。pCMV-E2F1-HA转染后PC3细胞凋亡率较未转染组显著上升[（7.10±0.003）% vs（2.66±0.001)%，P<0.05]，而pCMV-E2F1-HA与pcDNA3.1-TFDP3共转染后细胞凋亡率较pCMV-E2F1-HA组显著下降[（4.92±0.002）% vs（7.10±0.003）%，P<0.05]。结论：E2F1可增强TFDP3启动子的活性，增加TFDP3蛋白的表达，其可能通过此机制抑制E2F1诱导的前列腺癌细胞凋亡。

Objective : To evaluate the regulatory effects of E2F1 on transcription factor dimerization partner-3 ( TFDP3 ) expression and apoptosis in prostate cancer cells in vitro. Methods: A luciferase reporter construct driven by the human TFDP3 gene promoter, pGL3-TFDP3-promoter, was made through PCR and subcloning using the total DNA extracted from human prostate cancer PC3 cells. PC3 cells were transfected with pGL3-TFDP3-promoter and pCMV-E2F1-HA E2F1 expression vector, either alone or together. TFDP3 promoter activity and TFDP3 protein content in the transfectants were determined by dual luciferase assays and Western blotting analysis, respectively, 48 h after transfection, while apoptosis was analyzed by flow cytometry 24 h after transfection. Results: The luciferase activity was significantly higher in PC3 cells co-transfected with pGL3-TFDP3-promoter and pCMV-E2F1-HA than PC3 cells transfected with pGL3-TFDP3-promoter alone (1.14 vs 0.61, P<0.05). TFDP3 protein content in PC3 cells transfected with pCMV-E2F1-HA was 2.7 times higher than that in non-transfected cells ([0.24±0.03] vs [0.089±0.02], P<0.05). The proportion of apoptotic cells PC3 cells transfected with pCMV-E2F1-HA (7.1±0.003)% was significantly higher than that both in non-transfected PC3 cells ([2.66±0.001]% P<0.05) and in PC3 cells co-transfected with pcDNA3-TFDP3-promoter and pCMV-E2F1-HA ([4.92±0.002]% vs [7.1±0.003]%，P<0.05). Conclusion: E2F1 may enhance the TFDP3 promoter activity and upregulate TFDP3expression in prostate cancer cells. This finding suggests that E2F1 and TFDP3 may play a role in the survival/apoptosis in prostate cancer cells.

国家自然科学基金资助项目（No.81272619）