miR‑221 regulates proliferation and apoptosis of ovarian cancer cells by targeting BMF

Xie,Xinping; Huang,Yuxiu; Chen,Lihong; Wang,Jinhua

doi:10.3892/ol.2018.9446

miR‑221 regulates proliferation and apoptosis of ovarian cancer cells by targeting BMF

Authors:
- Xinping Xie
- Yuxiu Huang
- Lihong Chen
- Jinhua Wang
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Affiliations: Department of Gynecology and Obstetrics, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350004, P.R. China
Published online on: September 18, 2018 https://doi.org/10.3892/ol.2018.9446
Pages: 6697-6704

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Abstract

To observe the expression of microRNA‑221 (miR‑221) in ovarian cancer tissues and its effect and associated mechanism on proliferation and apoptosis in the ovarian cancer SKOV3 cell line. The expression of miR‑221 and B‑cell lymphoma 2 modifying factor (BMF) mRNA in ovarian cancer and para‑carcinoma tissues was detected by reverse transcription‑quantitative polymerase chain reaction, the expression of BMF was detected by western blot. MicroRNA.org online predicted that BMF was the possible target gene of miR‑221, and the regulatory association was validated by a dual‑luciferase reporter gene assay. SKOV3 cells were divided into 8 transfection groups: Anti‑miR‑negative control (NC); anti‑miR‑221; phosphorylated internal ribosome entry site 2 (pIRES2)‑blank, pIRES2‑BMF, small interfering (si)‑NC, si‑BMF, anti‑miR‑221+si‑BMF and anti‑miR‑221+pIRES2‑BMF groups. Cell proliferation was detected by EdU staining flow cytometry. The effect of transfection on cell apoptosis was detected by Annexin V/PI double staining, and the activity of caspase‑3 was detected by spectrophotometry. The effect of anti‑miR‑221 or pIRES2‑BMF transfection on SKOV3 cell proliferation was detected by MTT assay and flow cytometry, and the effect on apoptosis was detected by the Annexin V/PI double staining. Compared with para‑cancer tissues, the miR‑221 expression was significantly upregulated (P<0.001), the BMF mRNA expression was significantly downregulated (P<0.001), and the expression of BMF proteins was significantly downregulated in the ovarian cancer tissues. Dual‑luciferase reporter gene assay confirmed a targeted regulatory association between miR‑221 and BMF. The anti‑miR‑221 or pIRES2‑BMF transfection significantly upregulated BMF expression in SKOV3 cells, significantly decreased cell proliferation and significantly increased cell apoptosis. The overexpression of BMF may enhance the proapoptotic and proliferation‑inhibition effect of anti‑miR‑221 on SKOV3 cells. The transfection of si‑BMF significantly promoted cell proliferation, reduced cell apoptosis and attenuated the proapoptotic and proliferation‑inhibition effect of anti‑miR‑221 on cells. The expression of miR‑221 was significantly upregulated and the expression of BMF was significantly down‑regulated in ovarian cancer tissues. The overexpression of miR‑221 antagonized the apoptosis of ovarian cancer SKOV3 cell and promoted the cell proliferation by targeted inhibition of the expression of BMF, which may serve a role in the pathogenesis of ovarian cancer.

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