[摘 要] 目的：探讨长链非编码RNA 01694（LINC01694）调节miR-128-3p/端粒重复结合因子1（TERF1）轴对前列腺癌（PC）细 胞恶性生物学行为的影响。方法：收集2023年1月至2024年1月间在荆州市中心医院泌尿外科手术切除的20例PC组织及相应 癌旁组织，常规培养人PC细胞PC-3、DU145、LNCaP、C4-2和正常人前列腺上皮细胞RWPE-1。用Lipo6000TM转染试剂将 sh-LINC01694、sh-NC、miR-128-3p inhibitor、inhibitor-NC、miR-128-3pmimics、pcDNA和pcDNA-LINC01694转染LNCaP细胞，分 为Ctrl、sh-NC、sh-LINC01694、sh-LINC01694 + NC inhibitor、sh-LINC01694 + miR-128-3p inhibitor、pcDNA和pcDNA LINC01694 组。用qPCR法检测PC组织和细胞，以及各组LNCap细胞中LINC01694、miR-128-3p和TERF1 mRNA的表达，WB法检测各组 LNCaP细胞中TERF1、caspase-3、cyclin D1、E-cadherin、N-cadherin蛋白的表达，克隆形成实验、流式细胞术和Transwell小室实验 分别检测各组LNCaP细胞的增殖、迁移、侵袭能力，以及细胞凋亡情况。双萤光素酶报告基因实验、RNA pull-down实验和RNA 免疫共沉淀（RIP）实验验证LINC01694与miR-128-3p和TERF1与miR-128-3p的靶向结合关系。裸鼠LNCaP细胞移植瘤实验检 测敲减LINC01694对其移植瘤生长的影响。结果：LINC01694在PC组织、细胞中呈高表达（均P < 0.05），在LNCaP细胞中敲减 LINC01694可促进miR-128-3p、caspase-3、E-cadherin 蛋白的表达，抑制LINC01694、TERF1、cyclin D1、N-cadherin蛋白的表达， 抑制LNCaP细胞的增殖、迁移和侵袭能力，并促进其凋亡（均P < 0.05），这些作用均可被miR-128-3p inhibitor部分逆转（均 P < 0.05）。LINC01694可直接与miR-128-3p结合（P < 0.05），miR-128-3p可直接与TERF1mRNA结合（P < 0.05），说明LINC01694 可调控miR-128-3p/TERF1轴。敲减LINC01694可明显抑制裸鼠LNCaP细胞移植瘤的生长（P < 0.05）。结论：LINC01694通过 调节miR-128-3p/TERF1轴抑制LNCaP细胞的恶性生物学行为。

[Abstract] Objective: To investigate the effects of long non-coding RNA 01694 (LINC01694) regulating the microRNA-128-3p (miR-128-3p)/telomeric repeat binding factor 1 (TERF1) axis on the malignant biological behaviors of prostate cancer (PC) cells. Methods: Cancer tissues and corresponding adjacent tissues from 20 PC patients undergoing surgery at the Department of Urology, Jingzhou Central Hospital, between January 2023 and January 2024 were collected. Human PC cell lines (PC-3, DU145, LNCaP, C4-2) and normal human prostate epithelial RWPE-1 cells were routinely cultured. LNCaP cells were transfected with sh-LINC01694, sh-NC, miR-128-3p inhibitor, inhibitor-NC, miR-128-3pmimics, pcDNA, and pcDNA-LINC01694 using Lipo6000? transfection reagent. Cells were divided into the following groups: Ctrl, sh-NC, sh-LINC01694, sh-LINC01694 + NC inhibitor, sh-LINC01694 + miR-128-3p inhibitor, pcDNA, and pcDNA LINC01694 groups. The mRNA expression of LINC01694, miR-128-3p, and TERF1 in PC tissues and cells, as well as LNCap cells in each group, was detected by qPCR. The protein expression of TERF1, caspase-3, cyclin D1, E-cadherin, and N-cadherin in LNCaP cells of each group was detected by WB method. Clone formation assay, flow cytometry, and Transwell chamber assay were applied to detect proliferation, apoptosis, migration, and invasion of LNCaP cells, respectively. Dual luciferase reporter gene assay, RNA pull-down assay, and RNA-binding protein immunoprecipitation (RIP) assay were applied to verify the targeting binding relationship between LINC01694 and miR-128-3p as well as between TERF1 and miR-128-3p. Nude mouse LNCaP cell xenograft experiment was conducted to assess the effect of LINC01694 knockdown on tumor growth. Results: LINC01694 was highly expressed in PC tissues and cells (all P<0.05). Knockdown of LINC01694 in LNCaP cells promoted the protein expression of miR-128-3p, caspase-3, and E-cadherin, inhibited the protein expression of LINC01694, TERF1, cyclin D1, and N-cadherin, reduced cell proliferation, migration, and invasion, and promoted apoptosis (all P < 0.05). All these effects were partially reversed by the miR 128-3p inhibitor (all P < 0.05). LINC01694 could directly bind to miR-128-3p (P < 0.05), while miR-128-3p could directly bind to TERF1 mRNA (P < 0.05), indicating that LINC01694 regulates the miR-128-3p/TERF1 axis. Knockdown of LINC01694 significantly inhibited the growth of LNCaP cell xenografts in nude mice (P < 0.05). Conclusion: LINC01694 regulates the malignant biological behaviors of LNCaP cells through the miR-128-3p/TERF1 axis.

湖北省卫生健康委员会资助项目（No. WJ2019Q020）