MiR-499b-5p inhibits cervical cancer cell proliferation and induces apoptosis by targeting the Notch1 signaling pathway

H.-M. Liu, F. Ji, Y. Lu, S.-Y. Chen

Department of Gynaecology, First Affiliated Hospital, School of Medicine, Shihezi University, Xinjiang Uygur Autonomous Region, China. lhm5372j@163.com

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• Oncology

OBJECTIVE: We investigated the effect of miR-499b-5p on the tumorigenesis and development of cervical cancer by targeting the Notch1 signaling pathway to identify a new potential clinical target of cervical cancer.

PATIENTS AND METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to determine the mRNA expression levels of Notch1 and miR-499b-5p in cervical cancer tissues/cell lines. Cell counting kit-8 (CCK-8) assay, transwell assay, and flow cytometry were conducted to detect cell viability, cell migration, and cell apoptosis abilities. A Dual-Luciferase reporter assay was performed to test the binding site between miR-499b-5p and Notch1. An in vivo experiment was carried out using nude mice, and xenograft tumor models were established.

RESULTS: OD450 of the SiHa and HeLa cells of the miR-499b-5p agomir group was lower than that of the miR-499b-5p agomir-NC group. More apoptotic cells and fewer invasive cells were found in the former than in the latter. MiR-499b-5p inhibited the viability and migration of cervical cancer cells and promoted their apoptosis. Further detection of the Luciferase reporter gene confirmed the binding site of miR-499b-5p to Notch1. Western blot results showed that miR-499b-5p inhibited the expression of Notch1 and activated the expression of ChK2 and p-p38MAPK. Notch1 knockdown also inhibited the viability and migration of cervical cancer cells and promoted their apoptosis. MiR-499b-5p overexpression prevented the tumorigenesis and development of cervical cancer in xenograft tumor models.

CONCLUSIONS: MiR-499b-5p inhibits the proliferation of cervical cancer cells and induces their apoptosis by targeting the Notch1 signaling pathway.

 

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