Journal of Jilin University(Medicine Edition) ›› 2021, Vol. 47 ›› Issue (5): 1099-1107.doi: 10.13481/j.1671-587X.20210504

• Research in basic medicine • Previous Articles     Next Articles

Effects of miR-762 on proliferation and apoptosis of human tongue squamous cell carcinoma cells by targeting NCOR1 expression

Naigao TANG,Genjian ZHENG()   

  1. Department of Stomatology,First Affiliated Hospital,Hainan Medical College,Haikou 570100 China
  • Received:2021-01-22 Online:2021-09-28 Published:2021-10-26
  • Contact: Genjian ZHENG E-mail:hbwsl730@163.com

Abstract:

Objective: To investigate the targeting relationship between miR-762 and nuclear receptor corepressor 1(NCOR1)and its effects on the proliferation and apoptosis of human tongue squamous cell carcinoma(TSCC) cells, and to provide new molecular markers and targets for the clinical diagnosis and treatment of TSCC.

Methods

The expression levels of miR-762 and NCOR1 mRNA in cancer tissue and paracancerous normal tissue of 48 patients with TSCC were detected by Real-time fluorescence quantitative PCR(RT-qPCR), and the correlation between the their expressions was analyzed by Pearson correlation analysis. The expression levels of miR-762 and NCOR1 mRNA and proteins in human normal tongue epithelial cells Hacat and TSCC cells (TCA-8113, CAL-27, SCC-15 and SCC-9) were detected by RT-qPCR and Western blotting methods.miR -762 inhibitor or si-NCOR1 was transfected into the CAL-27 cells separately or simultaneously;the experiment was divided into blank control group, inhibitor-NC group, miR-762 inhibitor group, si-NC group, si-NCOR1 group and miR-762 inhibitor+si-NCOR1 group. The expression levels of miR-762 and NCOR1 mRNA in the CAL-27 cells were detected by RT-qPCR method. The cell proliferation activities in various groups were detected by MTT assay; the proportions of EdU positive staining cells in various groups were detected by EdU assay, and the apoptotic rates were detected by flow cytometry. The expression levels of NCOR1, cleaved-Caspase-3, Bax and Bcl-2 proteins in the cells in various groups were detected by Western blotting method. The targeted regulatory relationship between miR-762 and NCOR1 was verified by luciferase report assay.

Results

Compared with the paracancerous normal tissue and the normal human tongue epithelial cells Hacat,the expression levels of miR-762 in TSCC tissue and TSCC cells were significantly increased(P<0.01), while the expression levels of NCOR1 were significantly decreased(P<0.01);there was a negative correlation between their expressions in TSCC tissue(r=-0.711,P=0.003). Luciferase reporting assay confirmed that NCOR1 was the target gene of miR-762. Compared with blank control and inhibitor-NC groups, the proliferation activity of cells, the proportion of EdU positive cells and the expression level of Bcl-2 protein in miR-762 inhibitor group were significantly decreased (P<0.01),and the apoptotic rate and the expression levels of cleaved-Caspase-3 and Bax proteins were significantly increased (P<0.01);compared with miR-762 inhibitor group, the proliferation activity, the proportion of EdU positive cells and the expression level of Bcl-2 protein in miR-762 inhibitor+si-NCOR1 group were significantly increased (P<0.01),and the apoptotic rate and the expression levels of cleaved-Caspase-3 and Bax proteins were significantly decreased(P<0.01).

Conclusion

Inhibition of miR-762 expression can inhibit the proliferation and promote the apoptosis of TSCC cells by targeting and upregulating the expression of NCOR1.

Key words: tongue squamous cell carcinoma, miR-762, nuclear receptor corepressor 1, cell proliferation, apoptosis

CLC Number: 

  • R739.86