Li Yibing, Huo Jianing, Pan Xin, Wang Cuicui, Ma Xiaoxin
Department of Obstetrics and Gynecology, Shengjing Hospital of China Medical University, Shenyang, Liaoning, People's Republic of China.
Onco Targets Ther. 2018 Mar 6;11:1275-1284. doi: 10.2147/OTT.S154517. eCollection 2018.
Studies have shown that the microRNA miR-302 can affect the proliferation, migration and cell cycle progression of endometrial carcinoma (EC). miR-302 clusters have been shown to play an important role in the proliferation and differentiation of cancer cells and in their tumorigenicity.
In this study, we detected the expression of genes through quantitative reverse transcription polymerase chain reaction (qRT-PCR). We detected the expression of proteins through Western blot. The Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double-staining assay were used to detect the ability of miR-302b-3p/302c-3p/302d-3p to affect the cell apoptosis. The CCK-8 were used to detect the ability of miR-302b-3p/302c-3p/302d-3p to affect the cell proliferation. The Cell cycle analysis were used to detect the ability of miR-302b-3p/302c-3p/302d-3p to affect the cell cycle. Finally, the wound healing assay was used to detect the ability of miR-302b-3p/302c-3p/302d-3p to impact cell migration.
We found that miR-302b-3p/302c-3p/302d-3p of the miR-302 cluster was downregulated in EC, and it altered the epithelial-mesenchymal transition (EMT) process in the EC cell lines Ishikawa and HEC-1A. Western blot and the Annexin V- FITC/PI double-staining assay were used to detect the ability of miR-302b-3p/302c-3p/302d-3p to promote the apoptosis of Ishikawa and HEC-1A cells. In addition, qRT-PCR results showed that overexpression of miR-302b-3p/302c-3p/302d-3p significantly inhibited the expression of ZEB1, suppressed the expression of Bcl-2 and promoted the expression of BAX. The overexpression of miR-302b-3p/302c-3p/302d-3p inhibited the proliferation and migration of Ishikawa and HEC-1A cells. Cell cycle analysis showed that miR-302b-3p/302c-3p/302d-3p arrested cell cycle progression in the G0/G1 phase.
All results showed that miR-302b-3p/302c-3p/302d-3p can be used as a tumor suppressor in EC and is expected to be a new target for the treatment of EC.
研究表明,微小RNA miR - 302可影响子宫内膜癌(EC)的增殖、迁移及细胞周期进程。已证实miR - 302簇在癌细胞的增殖、分化及其致瘤性中发挥重要作用。
在本研究中,我们通过定量逆转录聚合酶链反应(qRT - PCR)检测基因表达。通过蛋白质免疫印迹法检测蛋白质表达。采用膜联蛋白V - 异硫氰酸荧光素(FITC)/碘化丙啶(PI)双染法检测miR - 302b - 3p/302c - 3p/302d - 3p影响细胞凋亡的能力。采用CCK - 8检测miR - 302b - 3p/302c - 3p/302d - 3p影响细胞增殖的能力。采用细胞周期分析检测miR - 302b - 3p/302c - 3p/302d - 3p影响细胞周期的能力。最后,采用伤口愈合试验检测miR - 302b - 3p/302c - 3p/302d - 3p影响细胞迁移的能力。
我们发现miR - 302簇中的miR - 302b - 3p/302c - 3p/302d - 3p在EC中表达下调,并且它改变了EC细胞系Ishikawa和HEC - 1A中的上皮 - 间质转化(EMT)过程。采用蛋白质免疫印迹法和膜联蛋白V - FITC/PI双染法检测miR - 302b - 3p/302c - 3p/302d - 3p促进Ishikawa和HEC - 1A细胞凋亡的能力。此外,qRT - PCR结果显示,miR - 302b - 3p/302c - 3p/302d - 3p的过表达显著抑制ZEB1的表达,抑制Bcl - 2的表达并促进BAX的表达。miR - 302b - 3p/302c - 3p/302d - 3p的过表达抑制了Ishikawa和HEC - 1A细胞的增殖和迁移。细胞周期分析表明,miR - 302b - 3p/302c - 3p/302d - 3p使细胞周期进程停滞在G0/G1期。
所有结果表明,miR - 302b - 3p/302c - 3p/302d - 3p可作为EC中的肿瘤抑制因子,有望成为EC治疗的新靶点。