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目的:探讨环磷腺苷效应元件结合蛋白(c AMP-response element binding protein,CREB)及磷酸化CREB(p-CREB)在前列腺癌细胞增殖中的作用。方法:采用组织芯片技术检测人正常前列腺、前列腺增生、不同分级前列腺癌组织中CREB及p-CREB的表达水平;免疫荧光及Western blotting检测人正常前列腺基质永生化细胞WPMY-1、前列腺癌细胞PC3及LNCap中CREB及p-CREB的表达水平;构建重组人源CREB质粒,并转染PC3及LNCap细胞,Western blotting及CCK-8试剂盒检测转染效率及细胞增殖水平的变化。结果:CREB及p-CREB在前列腺癌组织中表达率明显高于正常前列腺(96%vs 50%,88%vs 40%,P<0.05)和前列腺增生组织(96%vs 80%,88%vs 60%,P<0.05);CREB及p-CREB蛋白水平在前列腺癌PC3及LNCap细胞中表达量显著高于正常前列腺基质永生化细胞WPMY-1[PC3细胞:(5.10±0.62)vs(2.31±0.40)、(4.31±0.54)vs(2.02±0.38);LNCap细胞:(7.5±0.83)vs(2.31±0.40)、(6.15±0.69)vs(2.02±0.38),均P<0.05)];转染重组质粒CREB可上调前列腺癌细胞PC3及LNCap中增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达(均P<0.05),且转染后的PC3及LNCap细胞增殖水平明显增加(P<0.05)。结论:CREB及p-CREB在前列腺癌组织、体外培养前列腺癌细胞PC3及LNCap中高表达,并可上调PC3及LNCap中增殖相关基因PCNA的表达,提高细胞增殖水平。
Objective: To investigate the role of cAMP-response element binding protein (CREB) and phosphorylated CREB (p-CREB) in the proliferation of prostate cancer cells. METHODS: The expression of CREB and p-CREB in human normal prostate, benign prostatic hyperplasia and prostate cancer tissues were detected by tissue microarray. The expressions of WPMY-1 and PC3 in immortalized prostate stromal cells And CREB and p-CREB in LNCap. Recombinant human CREB plasmid was constructed and transfected into PC3 and LNCap cells. The transfection efficiency and cell proliferation were detected by Western blotting and CCK-8 kit. Results: The positive rates of CREB and p-CREB in prostate cancer were significantly higher than those in normal prostate (96% vs 50%, 88% vs 40%, P <0.05) and benign prostatic hyperplasia (96% vs 80%, 88% vs 60%, P <0.05). The expression of CREB and p-CREB protein in prostate cancer PC3 and LNCap cells were significantly higher than those in normal prostatic stromal cell immortalized cells [WPMY-1 [PC3 cells: (5.10 ± 0.62 vs (4.31 ± 0.54) vs (2.02 ± 0.38); LNCap cells: (7.5 ± 0.83) vs (2.31 ± 0.40), (6.15 ± 0.69) vs (2.02 ± 0.38, all P <0.05) The recombinant plasmid CREB upregulated the expression of proliferating cell nuclear antigen (PCNA) in PC3 and LNCap cells (all P <0.05), and the proliferation of PC3 and LNCap cells significantly increased (P <0.05 ). CONCLUSION: CREB and p-CREB are overexpressed in prostate cancer cells, in vitro cultured prostate cancer cells PC3 and LNCap, and can up-regulate the expression of PCNA and PCNA in PC3 and LNCap cells and increase the cell proliferation.