Objective: To investigate the regulation of long-chain non-coding RNA-AC024560.2 transfection on the expression of miR-30a-5p and its effect on proliferation and invasion of prostate cancer cells. Methods: qRT-PCR was used to detect the expression of AC024560.2 in 16 prostate cancer tissues and adjacent normal tissues, prostate cancer cell lines and normal prostate epithelial cells. The cells with the lowest expression amount were transfected, and the prostate cancer cells were divided into control group (transfected with negative control plasmid) and experimental group (transfected with plasmid carrying AC024560.2). Bioinformatics predicted possible target genes for AC024560.2. qRT-PCR was used to detect the expression of AC024560.2 and target genes in the transfected cells. Western blot was used to detect the expression of downstream target proteins. Cell proliferation and invasion were analyzed by MTS assay and Transwell invasion assay. Results: The expression levels of AC024560.2 in prostate cancer tissues and adjacent tissues were 1.95±0.22 and 3.87±0.23, respectively (t=6.09, P<0.01). Compared with normal prostate epithelial cells, the expression of AC024560.2 in prostate cancer cell lines was significantly decreased (P<0.05), and the most significant decrease was observed in C4-2B cell lines (P<0.01). Bioinformatics predictions showed that AC024560.2 bond to miR-30a-5p, and miR-30a-5p bond to SIRT1 mRNA. The expression of AC024560.2 in the experimental group increased significantly (P<0.01), the expression of miR-30a-5p decreased significantly (P<0.01), and the expression of SIRT1 mRNA and protein increased significantly (P<0.01). After transfection with AC024560.2, the cell proliferation ability of the experimental group was significantly decreased from day 2 (P<0.05). The invasive numbers of C4-2B cells in the control group and the experimental group were 130.90±14.54 and 43.77±10.01, respectively (t=4.94, P<0.01). Conclusions: AC024560.2 is lowly expressed in human prostate cancer, and may inhibit the proliferation and invasion of prostate cancer cells by regulating the expression of miR-30a-5p and SIRT1 genes. AC024560.2 may be a potential target for the treatment of prostate cancer.
目的： 探讨长链非编码RNA-AC024560.2转染对miR-30a-5p表达的调控作用及对前列腺癌细胞增殖和侵袭的影响。 方法： qRT-PCR检测2017年3月至2018年1月武汉市中心医院泌尿外科16例前列腺癌组织及癌旁组织、人前列腺癌细胞株及正常前列腺上皮细胞中AC024560.2表达量。以表达量最低的细胞为转染对象，前列腺癌细胞分为对照组（转染阴性对照质粒）和实验组（转染携带AC024560.2的质粒）。生物信息学预测AC024560.2可能的靶基因。qRT-PCR检测转染后细胞中AC024560.2和靶基因的表达量。Western印迹检测下游靶蛋白的表达。MTS法和Transwell侵袭实验分析细胞增殖和侵袭能力。 结果： 前列腺癌组织和癌旁组织中AC024560.2表达量分别为1.95±0.22和3.87±0.23（t=6.09，P<0.01）。与正常前列腺上皮细胞相比，前列腺癌细胞株中AC024560.2表达量均显著下降（P<0.05），其中C4-2B细胞株中下降最明显（P<0.01）。生物信息学预测显示，AC024560.2可靶向结合miR-30a-5p，miR-30a-5p可靶向结合沉默信息调节因子1（SIRT1）mRNA。实验组中AC024560.2表达量显著上升（P<0.01），miR-30a-5p表达量显著下降（P<0.01），SIRT1 mRNA和蛋白表达量显著上升（P<0.01）。转染AC024560.2后，实验组细胞增殖能力从第2天开始明显降低（P<0.05）。对照组和实验组C4-2B细胞的侵袭数分别为130.90±14.54和43.77±10.01（t=4.94，P<0.01）。 结论： AC024560.2在人前列腺癌中呈低表达，可能通过调控miR-30a-5p和SIRT1基因的表达，抑制前列腺癌细胞的增殖和侵袭能力。AC024560.2可能成为治疗前列腺癌的潜在靶点。.
Keywords: Long-chain non-coding RNA; Prostate cancer; SIRT1; microRNA.