Wang Y, Chen L, Guo Y L, Li G H, Ying C C
Department of Urology, the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China.
Zhonghua Yi Xue Za Zhi. 2019 Jul 9;99(26):2042-2046. doi: 10.3760/cma.j.issn.0376-2491.2019.26.008.
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. 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. The expression levels of AC024560.2 in prostate cancer tissues and adjacent tissues were 1.95±0.22 and 3.87±0.23, respectively (6.09, 0.01). Compared with normal prostate epithelial cells, the expression of AC024560.2 in prostate cancer cell lines was significantly decreased (0.05), and the most significant decrease was observed in C4-2B cell lines (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 (0.01), the expression of miR-30a-5p decreased significantly (0.01), and the expression of SIRT1 mRNA and protein increased significantly (0.01). After transfection with AC024560.2, the cell proliferation ability of the experimental group was significantly decreased from day 2 (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 (4.94, 0.01). 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.
探讨长链非编码RNA-AC024560.2转染对miR-30a-5p表达的调控及其对前列腺癌细胞增殖和侵袭的影响。采用qRT-PCR检测16例前列腺癌组织及癌旁正常组织、前列腺癌细胞系和正常前列腺上皮细胞中AC024560.2的表达。转染表达量最低的细胞，将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。采用qRT-PCR检测转染细胞中AC024560.2和靶基因的表达。采用Western blot检测下游靶蛋白的表达。通过MTS法和Transwell侵袭实验分析细胞增殖和侵袭能力。前列腺癌组织和癌旁组织中AC024560.2的表达水平分别为1.95±0.22和3.87±0.23（P=6.09，0.01）。与正常前列腺上皮细胞相比，前列腺癌细胞系中AC024560.2的表达明显降低（P<0.05），其中C4-2B细胞系降低最显著（P<0.01）。生物信息学预测显示AC024560.2与miR-30a-5p结合，miR-30a-5p与SIRT1 mRNA结合。实验组中AC024560.2的表达显著升高（P<0.01），miR-30a-5p的表达显著降低（P<0.01），SIRT1 mRNA和蛋白的表达显著升高（P<0.01）。转染AC024560.2后，实验组细胞增殖能力从第2天开始显著降低（P<0.05）。对照组和实验组中C4-2B细胞的侵袭数分别为130.90±14.54和43.77±10.01（P=4.94，0.01）。AC024560.2在人前列腺癌中低表达，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。AC024560.2可能是前列腺癌治疗的潜在靶点。
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