[摘 要] 目的：探讨环磷腺苷效应元件结合蛋白（CREB）对前列腺癌细胞恶性生物学行为的影响。方法：常规培养前列腺癌 细胞PC3，用转染试剂将过表达对照质粒，CREB过表达质粒（CREB-oe)、敲减对照序列（si-NC）和si-CREB序列转染至PC3细胞， 分为vector，CREB-oe、si-NC和si-CREB组。用划痕愈合实验、Transwell小室实验和流式细胞术分别检测各组细胞的迁移、侵袭 能力和细胞周期情况。用CRISPR/cas9技术构建CREB敲除的PC3细胞，用PC3细胞移植瘤实验检测CREB敲除对移植瘤生长 的影响。结果：在PC3细胞中成功地敲减或过表达了CREB（均P < 0.01），过表达或敲减CREB均可明显促进或抑制PC3细胞 的迁移、侵袭能力（均P < 0.01），过表达CREB可促使PC3细胞进入S期，而敲减CREB表达则使PC3细胞阻滞于G1期（均 P < 0.01）。成功地构建了CREB敲除PC3细胞，敲除CREB可明显抑制PC3细胞移植瘤的生长（P < 0.01）。结论：敲减或敲除 CREB能够抑制PC3细胞的迁移和侵袭，并使其阻滞于G1期，进而抑制移植瘤的生长。

[Abstract] Objective: To investigate the effects of cyclic adenosine monophosphate response element binding protein (CREB) on the malignant biological behaviors of prostate cancer PC3 cells. Methods: Prostate cancer PC3 cells were routinely cultured. The overexpression control plasmid (vector), CREB overexpression plasmid (CREB-oe), knockdown control sequence (si-NC), and si-CREB sequence were transfected into PC3 cells using transfection reagents, namely vector, CREB-oe, si-NC, and si-CREB groups. Scratch wound healing assay, Transwell chamber assay, and flow cytometry were performed to evaluate cell migration, invasion, and cell cycle distribution, respectively. PC3 cells with CREB knockout were constructed using CRISPR/Cas9 technology, and a xenograft tumor model was employed to evaluate the impact of CREB knockout on tumor growth in vivo. Results: CREB was successfully knocked down or overexpressed in PC3 cells (all P < 0.01). CREB overexpression significantly promoted, while its knockdown significantly inhibited the migration and invasion of PC3 cells (all P < 0.01). Overexpression of CREB promoted the transition of PC3 cells into the S phase, whereas knockdown of CREB induced G1 phase arrest (all P < 0.01). PC3 cells with CREB knockout were successfully constructed, and CREB knockout significantly inhibited the growth of PC3 cell-transplanted tumors (P < 0.01). Conclusion: Knockdown or knockout of CREB inhibits migration and invasion of PC3 cells and induces G1 phase arrest, thereby suppressing tumor growth.

国家自然科学基金（No. 81372747）