Department of Obstetrics and Gynecology, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang, China.
Eur Rev Med Pharmacol Sci. 2017 Nov;21(21):4810-4818.
Endometrial cancer is one of the three most common types of gynecologic cancer. The global incidence has increased in recent years. microRNAs (miRNAs) regulate numerous biological processes by binding to the 3'UTR of target mRNA to down-regulate protein synthesis.
Endometrial cancer patients received surgeries in our hospital were enrolled. MiR-15a-5p mimic or miR-15a-5p inhibitor was transfected into HEC-1-A cells by lentivirus. Colony formation assay was applied for detecting cell proliferation. Real-time PCR was performed to test miRNA and mRNA expression. Western blot was used to detect protein level. ChIP was adopted to test transcription activation. TOP/FOP was tested to determine Wnt signaling pathway activity. A dual-luciferase reporter assay was used to confirm miRNA target.
miR-15a-5p was decreased in endometrial cancer cells and tissues. miR-15a-5p overexpression restrained HEC-1-A cell proliferation and stemness. miR-15a-5p mimic transfection reduced mRNA and protein levels of the proteins which are related to cell proliferation and Wnt signaling pathway. MiR-15a-5p targeted a putative binding site in the 3'-UTR of Wnt3a gene, thus regulating Wnt signaling pathway. miR-15a-5p overexpression decreased Wnt3a protein expression. Wnt3a presented significant negative correlation with the miR-15a-5p level in endometrial cancer patients.
miR-15a-5p is a regulator of endometrial cancer cell proliferation by directly targeting Wnt3a to block Wnt signaling pathway.
子宫内膜癌是妇科三大常见癌症之一。近年来，全球发病率有所上升。微小 RNA（miRNA）通过与靶 mRNA 的 3'UTR 结合来下调蛋白质合成，从而调节许多生物过程。
我们医院收治的子宫内膜癌患者被纳入研究。通过慢病毒转染 miR-15a-5p 模拟物或 miR-15a-5p 抑制剂到 HEC-1-A 细胞中。集落形成实验用于检测细胞增殖。实时 PCR 用于检测 miRNA 和 mRNA 表达。Western blot 用于检测蛋白水平。ChIP 用于检测转录激活。TOP/FOP 用于检测 Wnt 信号通路活性。双荧光素酶报告基因实验用于验证 miRNA 靶标。
miR-15a-5p 在子宫内膜癌细胞和组织中减少。miR-15a-5p 过表达抑制 HEC-1-A 细胞增殖和干性。miR-15a-5p 模拟物转染降低了与细胞增殖和 Wnt 信号通路相关的蛋白质的 mRNA 和蛋白水平。miR-15a-5p 靶向 Wnt3a 基因 3'-UTR 中的一个假定结合位点，从而调节 Wnt 信号通路。miR-15a-5p 过表达降低了 Wnt3a 蛋白表达。Wnt3a 在子宫内膜癌患者中的表达与 miR-15a-5p 水平呈显著负相关。
miR-15a-5p 通过直接靶向 Wnt3a 阻断 Wnt 信号通路，成为子宫内膜癌细胞增殖的调节剂。
