Ostadrahimi Shiva, Abedi Valugerdi Manuchehr, Hassan Moustapha, Haddad Ghazal, Fayaz Shima, Parvizhamidi Monireh, Mahdian Reza, Fard Esfahani Pezhman
Department of Biochemistry, Pasteur Institute of Iran, Tehran, Iran.
Department of Laboratory Medicine, Department of Experimental Cancer Medicine, Karolinska Institutet Huddinge, 141 86 Stockholm, Sweden. Email:
Asian Pac J Cancer Prev. 2018 Aug 24;19(8):2305-2311. doi: 10.22034/APJCP.2018.19.8.2305.
Objective: Small non-coding RNA molecules are dysregulated in prostate cancer (PCa). In our previous study, downregulation of miR-1266 and miR-185 was demonstrated in PCa tissues and cell lines. The aim of the present study was to investigate whether miR-1266 and miR-185 are involved in the regulation of B-cell lymphoma (BCL) 2 and BCL2L1, respectively, and whether transfection of PCa cell lines with miR-1266 and miR-185 mimics can alter tumorigenic phenotypes. Methods: In order to investigate the regulation of BCL2 and BCL2L1 mRNA levels by miR-1266 and miR-185, respectively, a luciferase reporter assay was used. Real-time PCR was also used to analyze changes in the levels of BCL2 and BCL2L1 mRNAs in PCa cell lines following transfection with synthetic miR-1266 and miR-185. Cell apoptosis was determined by Annexin V protein expression analysis via flow cytometry. In addition to the MTT assay, a cell proliferation assay was performed. Result: A luciferase assay confirmed that the BCL2 and BCL2L1 genes may be targeted by miR-1266 and miR-185, respectively, through binding to their 3′UTR regions. Transfection of PC3 and DU145 cells with miR-1266 and miR-185 induced apoptosis and reduced proliferation, which also revealed an inverse correlation with BCL2 and BCL2L1 gene expression in the treated cells. Conclusion: Our data suggests that miR-1266 and miR-185 may be novel candidates for further research in PCa treatment through the anti-apoptotic pathway.
小非编码RNA分子在前列腺癌（PCa）中表达失调。在我们之前的研究中，已证实在PCa组织和细胞系中miR - 1266和miR - 185表达下调。本研究的目的是分别研究miR - 1266和miR - 185是否参与B细胞淋巴瘤（BCL）2和BCL2L1的调控，以及用miR - 1266和miR - 185模拟物转染PCa细胞系是否能改变致瘤表型。方法：为了分别研究miR - 1266和miR - 185对BCL2和BCL2L1 mRNA水平的调控，采用了荧光素酶报告基因检测法。还使用实时PCR分析用合成的miR - 1266和miR - 185转染PCa细胞系后BCL2和BCL2L1 mRNA水平的变化。通过流式细胞术分析膜联蛋白V蛋白表达来确定细胞凋亡。除MTT法外，还进行了细胞增殖检测。结果：荧光素酶检测证实BCL2和BCL2L1基因可能分别被miR - 1266和miR - 185靶向，通过与它们的3′UTR区域结合。用miR - 1266和miR - 185转染PC3和DU145细胞诱导了细胞凋亡并降低了增殖，这也显示出与处理后细胞中BCL2和BCL2L1基因表达呈负相关。结论：我们的数据表明，miR - 1266和miR - 185可能是通过抗凋亡途径在PCa治疗中进一步研究的新候选物。
