Department of Oncology, The Third Xiangya Hospital of Central South University, Changsha, Hunan, China (mainland).
Med Sci Monit. 2019 Feb 11;25:1122-1132. doi: 10.12659/MSM.912148.
BACKGROUND The inhibitor of apoptosis, B-cell lymphoma 2 (Bcl-2), is encoded by the BCL2 gene. Previous studies have shown that microRNAs are downregulated in prostate cancer. This study aimed to investigate the role of microRNA-205 and microRNA-338-3p and cell apoptosis in prostate carcinoma tissue and the LNCaP human prostate adenocarcinoma cell line by directly targeting the BCL2 gene and Bcl-2 protein expression. MATERIAL AND METHODS Bioinformatics methods predicted the target genes of miR-205 and miR-338-3p, which were validated by a luciferase assay. Immunohistochemistry was used to detect Bcl-2 protein expression in 30 samples of prostate carcinoma tissue and 30 matched samples of normal prostate. The normal prostate epithelial cell line, RWPE-1, and LNCaP human prostate adenocarcinoma cells studied in vitro. BCL2 mRNA expression and Bcl-2 protein expression were determined by quantitative polymerase chain reaction (q-PCR) and Western blot, respectively. Cell apoptosis was measured by flow cytometry using annexin V, fluorescein isothiocyanate, and phycoerythrin (annexin V-FITC/PE). RESULTS TargetScan Human 7.2 predicted that the structures of miR-205 and miR-338-3p had a binding site on the proto-oncogene, BCL2, which was verified by a luciferase assay. The expression of miR-205 and miR-338-3p were significantly downregulated in prostate carcinoma tissues and LNCaP cells when compared with normal controls. BCL2 expression was significantly inhibited by overexpression of miR-205 and miR-338-3p in LNCaP cells. CONCLUSIONS The results of this study showed that miR-205 and miR-338-3p downregulated the expression of the BCL2 gene and decreased apoptosis in prostate carcinoma.
凋亡抑制因子 B 细胞淋巴瘤 2(Bcl-2)由 BCL2 基因编码。先前的研究表明,前列腺癌中 microRNA 的表达下调。本研究旨在通过直接靶向 BCL2 基因和 Bcl-2 蛋白表达,探讨 microRNA-205 和 microRNA-338-3p 与细胞凋亡在前列腺癌组织和 LNCaP 人前列腺腺癌细胞系中的作用。
生物信息学方法预测 miR-205 和 miR-338-3p 的靶基因,并通过荧光素酶报告基因实验进行验证。免疫组织化学法检测 30 例前列腺癌组织和 30 例配对正常前列腺组织中 Bcl-2 蛋白的表达。体外研究正常前列腺上皮细胞系 RWPE-1 和 LNCaP 人前列腺腺癌细胞。通过定量聚合酶链反应(q-PCR)和 Western blot 分别检测 BCL2 mRNA 表达和 Bcl-2 蛋白表达。用 Annexin V、异硫氰酸荧光素和藻红蛋白(annexin V-FITC/PE)通过流式细胞术测量细胞凋亡。
TargetScan Human 7.2 预测 miR-205 和 miR-338-3p 的结构在原癌基因 BCL2 上有一个结合位点,通过荧光素酶报告基因实验得到验证。与正常对照相比,miR-205 和 miR-338-3p 在前列腺癌组织和 LNCaP 细胞中的表达明显下调。在 LNCaP 细胞中过表达 miR-205 和 miR-338-3p 可显著抑制 BCL2 表达。
本研究结果表明,miR-205 和 miR-338-3p 下调 BCL2 基因的表达,减少前列腺癌的凋亡。