Pifu-xingbing zhenliaoxue zazhi (Aug 2021)

miR-196-5p promotes cutaneous squamous cell carcinoma proliferation, invasion, and EMT by activating the JAK/STAT pathway through targeting to HOXA5

  • Yonghong ZHANG,
  • Xuelian DAI,
  • Cuntao LI,
  • Yuhong ZHANG

DOI
https://doi.org/10.3969/j.issn.1674-8468.2021.04.002
Journal volume & issue
Vol. 28, no. 4
pp. 254 – 260,267

Abstract

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Objective: To investigate the effects of miR-196-5p on the proliferation, invasion and EMT of skin squamous cell carcinoma and its potential mechanism. Methods: Quantitative real-time PCR (RT-qPCR) was used to detect the expression of miR-196-5p in normal paracancer tissues, skin squamous cell carcinoma tissues, A431 cell and HaCaT cell. After the expression of miR-196-5p was overexpressed or down-regulated, the effects of miR-196-5p on the proliferation ability of A431 cells were detected by CCK-8 experiment. The effects of miR-196-5p on the invasion ability of A431 cells were detected by transwell. Western blot was used to detect the effects of miR-196-5p on the expression of E-cadherin and N-cadherin in A431 cells. TargetScan predicted the action sites of miR-196-5p and HOXA5. The relationship between miR-196-5p and HOXA5 was analyzed by dual luciferase reporter gene assay. After up-regulating HOXA5 expression in A431 cells, the effects of HOXA5 on A431 cell proliferation, invasion and EMT were detected. The effects of down-regulation of HOXA5 on the JAK/STAT pathway was detected by western blot. After AG490 was applied to cells, the effects of down-regulation of HOXA5on A431 cell proliferation、invasion and EMT were detected. Results: The expression level of miR-196-5p in cutaneous squamous cell carcinoma tissues was significantly higher than that in paracancer tissues (t=6.32, P=0.024), HOXA5 expression significantly decreased (t=7.60, P=0.017). Compared with HaCaT cells, miR-196-5p expression was significantly increased (t=18.58, P=0.003), HOXA5 expression significantly decreased in A431 cells (t=19.72, P=0.003). Overexpression of miR-196-5p promoted proliferation, invasion and EMT of A431 cells, while down-regulation of miR-196-5p had an opposite effect. miR-196-5p targeted and negatively regulated HOXA5.Overexpression of HOXA5 inhibited the proliferation, invasion and EMT of A431 cells. Down-regulation of HOXA5 activated the JAK/STAT pathway in A431 cell. AG490 downregulated the promotion of HOXA5 on A431 cell proliferation, invasion and EMT of A431 cell. Conclusion: miR-196-5p activates the JAK/STAT pathway by targeting HOXA5 to promote cutaneous squamous cell carcinoma proliferation, invasion and EMT.

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