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MicroRNA differential expression spectrum and microRNA-125a-5p inhibition of laryngeal cancer cell proliferation.

ABSTRACT: The present study aimed to screen and analyze the differential expression spectrum of microRNA (miRNA) between laryngeal cancer tissue and surrounding normal laryngeal mucosa in order to provide an indication for further study to determine the role of miRNA in the initiation and development of laryngeal cancer. A total of 42 pairs of specimens of laryngeal carcinoma tissues and adjacent normal laryngeal mucosa were collected. A total of 10 pairs of specimens were randomly selected for miRNA microarray gene chip analysis, and the remaining 32 pairs of specimens were used for reverse transcription-quantitative polymerase chain reaction (RT-qPCR) verification to identify miRNA that were differentially expressed in laryngeal cancer tissues. Methylthiazolyldiphenyl-tetrazolium bromide and clone formation assays were utilized to elucidate the physiological relevance of the miRNA miR-125a-5p on the proliferation of laryngeal cancer human epithelial type 2 (Hep2) cells. Results demonstrated that the expression levels of six miRNA were significantly downregulated in laryngeal carcinoma tissue, as identified by gene chip analysis and RT-qPCR (P<0.05). The six miRNA included let-7f-5p, miR-10a-5p, miR-125a-5p, miR-144-3p, miR-195-5p and miR-203. Compared with the control group, the proliferative ability of laryngeal cancer Hep2 cells was inhibited in a transfected miR-125a-mimics group. In contrast, proliferation was promoted in a transfected miR-125a-inhibitor group. In conclusion, the results of gene chip analysis were consistent with that of RT-qPCR. Results demonstrated that miRNA in laryngeal cancer and normal laryngeal mucosa exhibited evident differential expression, which may contribute to the laryngeal cancer incidence and invasion. miR-125a was able to inhibit the proliferation of Hep2 laryngeal cancer cells and, therefore, may serve as a novel target for laryngeal cancer biological therapy.

SUBMITTER: Yao XD

PROVIDER: S-EPMC5525897 | biostudies-other | 2017 Aug

REPOSITORIES: biostudies-other

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Project description:As a member of the integrin family, integrin α3β1 (ITGA3) has been linked to intercellular communication and serves an important role in the signaling among cells and the extracellular matrix. MicroRNA (miR)‑199a‑5p has been demonstrated to be related to the pathogenesis and progression of multiple malignant diseases. However, the biological functions of miR‑199a‑5p and ITGA3 in colorectal cancer (CRC) have rarely been reported. The aim of the present study was to explore the roles of miR‑199a‑5p and ITGA3 in CRC. Immunohistochemistry staining and western blotting were applied to detect the protein expression of ITGA3 in CRC tissues and cells. Reverse transcription‑quantitative PCR was performed to investigate the expression of miR‑199a‑5p and ITGA3 mRNA. HCT‑116 cells were transfected with miR‑199a‑5p mimics, mimics control, short hairpin RNA targeting ITGA3, or pcDNA‑ITGA3 for the functional experiments. Dual luciferase reporter assay was applied to confirm whether miR‑199a‑5p targeted the 3' untranslated region (3'UTR) of ITGA3. The MTT, Transwell and wound healing assays were used to evaluate the proliferation, invasion and migration of CRC cells. Immunofluorescence assay was used to monitor the epithelial‑mesenchymal transition (EMT) biomarker expression. The results demonstrated downregulation of miR‑199a‑5p and upregulation of ITGA3 in CRC tissues and cell lines. miR‑199a‑5p mimics and knockdown of ITGA3 suppressed the proliferation, invasion and migration of CRC cells. Bioinformatics analysis and luciferase reporter assay indicated that miR‑199a‑5p targeted the 3'UTR of the ITGA3 transcript, and overexpression of ITGA3 reversed the tumor‑suppressive effects of miR‑199a‑5p elevation. In addition, the immunofluorescence assay suggested that miR‑199a‑5p mimics suppressed the EMT of CRC cells, whereas the overexpression of ITGA3 restored this effect. In conclusion, miR‑199a‑5p may act as a tumor suppressor by targeting and negatively regulating ITGA3 in CRC.

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MicroRNA differential expression spectrum and microRNA-125a-5p inhibition of laryngeal cancer cell proliferation.

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