miR‑34c‑5p targets Notch1 and suppresses the metastasis and invasion of cervical cancer

Abstract

Micro (mi)RNAs are crucial participants in the progression of cervical cancer (CC). Growing evidence indicates that miRNA (miR)‑34c‑5p is a pivotal tumor suppressor in numerous types of cancer and its functions in CC require further investigating. The present study demonstrated that there was a decreased level of miR‑34c‑5p in CC‑associated cell lines compared with healthy control samples. It also demonstrated that miR‑34c‑5p targeted Notch1 and suppressed CC progression. Dual‑luciferase reporter assays verified the targeted relationship of miR‑34c‑5p and Notch1. The expression of Notch1 in HeLa cells was markedly reduced following miR‑34c‑5p overexpression and the proliferation, migration and invasion of HeLa cells were reduced although apoptosis was accelerated. However, overexpression of miR‑34c‑5p was reversed following the addition of Notch1, which supported the finding of the targeted relationship between miR‑34c‑5p and Notch1. Flow cytometry demonstrated that miR‑34c‑5p inhibited the proliferation of HeLa cells while accelerating apoptosis. The present study concluded that miR‑34c‑5p was a tumor suppressor in CC and may be a novel measure for the future treatment of CC.

Figure 1.

miR-34c-5p expression is downregulated in human cervical carcinoma tissues and cells. (A) RT-qPCR assay was used to detect the miR-34c-5p expression in 30 pairs of human cervical carcinoma tissues and control. (B) The miR-34c-5p expression in cervical carcinoma cell lines and normal cervical cell line were determined via RT-qPCR. ^##P<0.01. miR, microRNA; RT-qPCR, reverse transcription-quantitative PCR.

Figure 2.

miR-34c-5p inhibits the capacities of cervical carcinoma cell proliferation. (A) miR-34c-5p expression levels in mimics/miR-NC and inhibitors/anti-NC transfected cells were determined by reverse transcription-quantitative PCR. (B) HeLa cell viability was assessed with a CCK-8 assay. (C and D) An EdU assay was used to analyze the proliferation rates of HeLa cells. Magnification, ×200. ^#P<0.05, ^##P<0.01. miR, microRNA; NC, negative control; OD, optical density.

Figure 3.

Cell proliferation, apoptosis and cell cycle were detected by flow cytometry. ^#P<0.05. NC, negative control; miR, microRNA.

Figure 4.

miR-34c-5p inhibits the capacities of cervical carcinoma cell migration and invasion. (A) Transwell assays were performed to analyze the migration and the invasion capabilities of HeLa cells. Magnification, ×200. (B and C) Statistical results of the Transwell assays. ^#P<0.05, ^##P<0.01. NC, negative control; miR, microRNA; OD, optical density.

Figure 5.

Notch1 is a target of miR-34c-5p. (A) The 3′-UTR of Notch1 mRNA encompasses a highly conserved binding site for miR-34c-5p matching. (B) 293T cells were co-transfected with miR-34c-5p mimics and WT Notch1 mRNA 3′-UTR/mut Notch1 mRNA 3′-UTR. Luciferase activity was analyzed after 24 h of transfection. (C) mRNA expression levels of Notch1 were determined in HeLa cells (D and E) The protein expression levels of Notch1 were detected in HeLa cells. ^##P<0.01. miR, microRNA; UTR, untranslated regions; WT, wild-type; hsa, Homo sapiens; MUT, mutant; NC, negative control.