目的：探讨长链非编码RNA（long non-coding RNA，lncRNA）GTSE1-AS1在前列腺癌组织中的表达及其影响LNCaP细胞增殖和侵袭的机制。方法: 收集2017年11月至2018年12月郑州大学附属洛阳中心医院泌尿外科手术切除的68例前列腺癌患者的癌和癌旁组织标本，以及前列腺癌细胞系LNCaP、PC-3、C4-2B、22Rv1、DU-145和正常前列腺滤泡上皮细胞RWPE-1，用qPCR法检测癌组织及细胞系中GTSE1-AS1的表达水平。将GTSE1-AS1过表达质粒（实验组）或阴性对照质粒（对照组）转染至LNCaP细胞，用MTT法、Transwell小室法分别检测过表达GTSE1-AS1对LNCaP细胞增殖和侵袭能力的影响。通过生物信息学方法预测和双荧光素报告基因实验验证GTSE1-AS1与miR-324-3p、F框/WD重复域蛋白7（FBXW7）三者的靶向关系，用qPCR法和WB法检测过表达GTSE1-AS1对下游基因及蛋白表达的影响。结果: GTSE1-AS1在前列腺癌组织中的表达水平显著低于癌旁组织（P<0.01），GTSE1-AS1在前列腺癌细胞系中的表达显著低于RWPE-1细胞（P<0.05或P<0.01）。过表达GTSE1-AS1可显著抑制LNCaP细胞的增殖和侵袭能力（P<0.05 或 P<0.01）。双荧光素报告基因实验证实GTSE1-AS1互补结合miR-324-3p，miR-324-3p互补结合FBXW7。过 表 达 GTSE1-AS1可显著降低LNCaP细胞中miR-324-3p 的表达水平（P<0.01），促进 FBXW7 mRNA和蛋白的表达（均P<0.01）。结论: GTSE1-AS1在前列腺癌组织及细胞系中均低表达，过表达GTSE1-AS1可抑制LNCaP细胞的增殖和侵袭，其作用机制可能是通过抑制miR-324-3p表达从而促进FBXW7基因的表达。

Objective: To explore the expression of long non-coding RNA (lncRNA) GTSE1-AS1 in prostate cancer tissues and the mechanism that affects the proliferation and invasion of LNCaP cells. Methods: From November 2017 to December 2018, 68 pairs of prostate cancer tissue and para-cancerous tissue specimens were resected from prostate cancer patients at the Department of Urology of Luoyang Central Hospital Affiliated to Zhengzhou University; in addition, prostate cancer cell lines LNCaP, PC-3, C4-2B, 22Rv1,DU-145 and normal prostate follicular epithelial RWPE-1 cells were also chosen for this study. qPCR was used to detect the expression level of GTSE1-AS1 in cancer tissues and cell lines. The GTSE1-AS1 over-expression plasmid (experimental group) and negative control plasmid (control group) were respectively transfected into LNCap cells. MTT assay and Transwell chamber method were used to detect the effect of GTSE1-AS1 over-expression on the proliferation and invasion ability of LNCaP cells, respectively. The targeting relationship among GTSE1-AS1 and miR-324-3P as well as FBXW7 (F-frame/WD repeat domain protein 7) was verified by bioinformatics tools and dual-luciferin reporter gene assay. The effect of GTSE1-AS1 over-expression on downstream gene and protein expression was detected by qPCR and WB assay. Results: The expression level of GTSE1-AS1 in prostate cancer tissues was significantly lower than that in para-cancerous tissues (P<0.01), and the expression of GTSE1-AS1 in prostate cancer cell lines was significantly lower than that in RWPE-1 cells (P<0.05 or P<0.01). Over-expression of GTSE1-AS1 significantly inhibited the proliferation and invasion (P<0.05 or P<0.01) of LNCaP cells. Dual-luciferin reporter gene assay confirmed the complementary binding between GTSE1-AS1 and miR-324-3p as well as between miR-324-3p and FBXW7. Over-expression of GTSE1-AS1 significantly reduced the expression of miR-324-3p in LNCaP cells (P<0.01), and promoted the mRNA and protein expressions of FBXW7 (all P<0.01). Conclusion: GTSE1-AS1 is under-expressed in prostate cancer tissues and cell lines. Over-expression of GTSE1-AS1 can inhibit the proliferation and invasion of LNCaP cells, the mechanism of which may be related with the inhibition of miR-324-3p to further promote FBXW7 expression.

河南省医学科技攻关计划资助项目（No. 2018020895）