Dysregulation of micro-143-3p and BALBP1 contributes to the pathogenesis of the development of ovarian carcinoma.
Dysregulation of micro-143-3p and BALBP1 contributes to the pathogenesis of the development of ovarian carcinoma.
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DOI：
10.3892/or.2016.5148
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发表时间：
2016-12
期刊：
影响因子：
4.2
通讯作者：
Hongyan Zhang;Wanbin Li
中科院分区：
文献类型：
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作者：
Hongyan Zhang;Wanbin Li
本研究的目的是确定mir-143-3p与rala结合蛋白1 （RALBP1）之间的关联，以及它们在调节卵巢癌发展中的作用。RALBP1过表达诱导卵巢癌细胞凋亡，发展为卵巢癌。采用硅分析和荧光素酶测定来确定RALBP1是否是mir-143-3p的靶标。随后采用real - time PCR和western blotting检测不同处理组mir-143-3p、RALBP1 mRNA和蛋白的表达水平，并采用MTT法和流式细胞术检测不同处理组细胞的活力和凋亡情况。我们通过计算分析确定RALBP1是miR-143-3p的靶基因，转染野生型RALBP1和RALBP1 siRNA的细胞的荧光素酶活性远低于scramble对照组，而转染突变型RALBP1的细胞的荧光素酶活性与scramble对照组相似。real-time PCR和western blot结果显示，卵巢癌患者miR - 143-3p水平明显低于正常对照组，而RALBP1 mRNA和蛋白的表达在卵巢癌患者中明显过表达。此外，与scramble对照组相比，转染miR-143-3p模拟物和RALBP1 siRNA的细胞中RALBP1 mRNA和蛋白水平下调，而转染miR-143-3p抑制剂后，RALBP1 mRNA和蛋白水平明显上调。此外，转染miR-143-3p模拟物和RALBP1 siRNA后，细胞活力受到抑制，而转染miR-143-3p抑制剂后，细胞活力明显增强。转染miR-143-3p模拟物和RALBP1 siRNA后，细胞凋亡促进，转染miR-143-3p抑制剂后，细胞凋亡明显抑制。这些发现支持miR-143-3p的下调与卵巢癌风险降低相关。
The objective of the present study was to identify the association between mir-143-3p and RalA-binding protein 1 (RALBP1), and their roles in regulating the development of ovarian cancer. Overexpression of RALBP1 induced apoptosis of the ovarian cancer cells, and developed ovarian cancer. In silico analysis and luciferase assay were used to identify whether RALBP1 was the target of mir-143-3p. Subsequently, real‑time PCR and western blotting were used to determine the expression level of mir-143-3p, RALBP1 mRNA and protein in different groups, furthermore, MTT assay and flow cytometry were used to detect the viability and apoptosis of cells in different treatment groups. We identified RALBP1 as a target gene of miR-143-3p using computational analysis, and the luciferase activity of cells transfected with wild-type RALBP1 and RALBP1 siRNA were much lower than the scramble control, however, the luciferase activity of cells transfected with mutant RALBP1 was similar with scramble control. The real-time PCR and western blot results suggested that the miR‑143-3p level was markedly lower in participants with ovarian cancer compared with normal control, while the expression of RALBP1 mRNA and protein were evidently overexpressed in participants with ovarian cancer compared with normal control. Furthermore, the RALBP1 mRNA and protein level in cells transfected with miR-143-3p mimics and RALBP1 siRNA were downregulated, while notably upregulated subsequent to transfection with miR-143-3p inhibitor, when compared with scramble control. Additionally, the viability of cells were inhibited following transfection with miR-143-3p mimics and RALBP1 siRNA, while notably promoted subsequent to transfection with miR-143-3p inhibitor. Apoptosis of cells were promoted following transfection with miR-143-3p mimics and RALBP1 siRNA, while notably inhibited subsequent to transfection with miR-143-3p inhibitor. These findings provide support that downregulation of the miR-143-3p is associated with a decreased risk of ovarian cancer.