MiRNA-497 regulates cell growth and invasion by targeting cyclin E1 in breast cancer.
MiRNA-497 regulates cell growth and invasion by targeting cyclin E1 in breast cancer.
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miRNA-497 通过靶向细胞周期蛋白 E1 调节乳腺癌细胞生长和侵袭
DOI：
10.1186/1475-2867-13-95
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发表时间：
2013-10-10
影响因子：
5.8
通讯作者：
Fang L
中科院分区：
文献类型：
--
作者：
Luo Q;Li X;Gao Y;Long Y;Chen L;Huang Y;Fang L
MicroRNA是一类内源性单链非编码RNA，参与许多重要的生理和病理过程。本研究的目的是检测人乳腺癌中 miR-497 的表达水平及其在 MDA-MB-231 乳腺癌细胞中的功能。采用定量聚合酶链反应测定40例乳腺癌标本及癌旁正常乳腺组织中miR-497的表达水平。采用MTT测定、集落形成测定、伤口愈合测定、Transwell测定和细胞周期测定来探索miR-497在MDA-MB-231乳腺癌细胞中的潜在功能。进行双荧光素酶报告基因测定来分析 miR-497 假定靶标的调节，并使用蛋白质印迹测定来验证双荧光素酶结果。乳腺癌标本中miR-497的表达量低于癌旁正常组织(P<0.05)。 miR-497 的过度表达抑制细胞生长，抑制细胞迁移和侵袭，并导致 G1 期停滞。双荧光素酶报告基因检测显示 miR-497 结合细胞周期蛋白 E1 的 3'-非翻译区 (3'-UTR)，表明细胞周期蛋白 E1 是 miR-497 的直接靶标。蛋白质印迹测定证实，miR-497 的过度表达降低了细胞周期蛋白 E1 蛋白水平。 MiR-497 可能充当乳腺癌的抑癌基因。在细胞中过表达 miR-497 后，观察到细胞生长受到抑制、细胞迁移和侵袭受到抑制以及 G1 细胞周期停滞，这可能是通过靶向细胞周期蛋白 E1 实现的。这些结果表明 miR-497 可被视为开发乳腺癌治疗的治疗靶点。
MicroRNAs are a class of endogenous single strand non-coding RNAs that are involved in many important physiological and pathological processes. The purpose of this study was to examine the expression levels of miR-497 in human breast cancer and its function in MDA-MB-231 breast cancer cells. Quantitative polymerase chain reaction was used to measure the expression levels of miR-497 in 40 breast cancer specimens and adjacent normal breast tissues. MTT assays, colony formation assays, wound healing assays, transwell assays and cell cycle assays were used to explore the potential function of miR-497 in MDA-MB-231 breast cancer cells. Dual-luciferase reporter assays were performed to analyze the regulation of putative target of miR-497, and western blot assays were used to validate the dual-luciferase results. The expression of miR-497 in breast cancer specimens was lower than adjacent normal tissues (P < 0.05). Overexpression of miR-497 inhibited cellular growth, suppressed cellular migration and invasion, and caused a G1 arrest. Dual-luciferase reporter assays showed that miR-497 binds the 3′-untranslated region (3′-UTR) of cyclin E1, suggesting that cyclin E1 is a direct target of miR-497. Western blot assays confirmed that overexpression of miR-497 reduced cyclin E1 protein levels. MiR-497 may act as a tumor suppressor gene in breast cancer. Inhibited cellular growth, suppressed cellular migration and invasion, and G1 cell cycle arrest were observed upon overexpression of miR-497 in cells, possibly by targeting cyclin E1. These results indicate miR-497 could be considered a therapeutic target for the development of treatment for breast cancer.