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MiR-30a-5p Inhibits Epithelial Mesenchymal Transition Function in Human Cervical Cancer Cells by Targeting Ubiquitin-specific Protease 22 / 中山大学学报(医学科学版)
Journal of Sun Yat-sen University(Medical Sciences) ; (6): 698-705, 2019.
Article in Chinese | WPRIM | ID: wpr-817718
ABSTRACT
@# 【Objective】To investigate the inhibitory effect and mechanism of microRNA-30a-5p(miR-30a-5p)on epithelial mesenchymal transition in cervical cancer Hela cells.【Methods】Hela cervical cancer cell lines were transfected with miR-30a-5p mimics or negative control mimic,respectively,as 30a-5p or NC group. Control group was established with untreated Hela cervical cancer cells. miR-30a-5p content in each group was detected by RT-PCR assay. Transwell assay was used to detect the invasion ability of the 3 groups. Western-blot assay was used to detect the expressions of N- cadherin,α-Catenin and ubiquitin specific processing peptidase 22(USP22)protein in the 3 groups. Prediction target genes of miR-30a-5p by bioinformatics methods. Antagonistic effect of USP22 over-expression on miR-30a-5p inhibi⁃tion of EMT was detected by western blot assay. The relationship between miR-30a-5p and USP22 was detected by dual luciferase assay. Subcutaneous transplantation tumor model established,and the effect of miR-30a-5p in vivo was ob⁃ served.【Results】The miR-30a-5p intracellular quantity in 30a-5p group Hela cells was up-regulated,and the expres⁃ sion level of miR-30a-5p was 853.82(862.26~843.11)times higher than that of Control group(P<0.01). The number of invasive cells in 30a-5p group was 8.17(8.32~8.03),which was significantly lower than that of Control group 62.33 (63.52~60.19)(P<0.01). USP22 may be the target gene of miR-30a-5p. In 30a-5p group,the intracellular quantity of N-cadherin protein was decreased,the intracellular quantity of α-Catenin protein was increased,and the intracellular quantity of USP22 protein was decreased. The intracellular quantity of N-cadherin protein in 30a-5p group was down-reg⁃ ulated,the intracellular quantity of α-Catenin protein was increased,and the intracellular quantity of USP22 protein was reduced. After USP22 over-expression,the intracellular quantity of N-cadherin protein in cervical cancer cells of 30a-5p group was up-regulated,and the intracellular quantity of α-Catenin protein were down-regulated. Dual luciferase assay showed that USP22 is a downstream target gene of miR-30a-5p(P<0.01). Subcutaneous transplantation tumors in 30a- 5p group were significantly smaller than those in Control group.【Conclusion】miR-30a-5p may inhibit the expression of EMT related protein through the downstream target gene USP22,and the epithelial mesenchymal transition function of cervical cancer Hela cells.

Full text: Available Index: WPRIM (Western Pacific) Type of study: Prognostic study Language: Chinese Journal: Journal of Sun Yat-sen University(Medical Sciences) Year: 2019 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Type of study: Prognostic study Language: Chinese Journal: Journal of Sun Yat-sen University(Medical Sciences) Year: 2019 Type: Article