LncRNA HOXA11-AS Promotes Proliferation and Cisplatin Resistance of Oral Squamous Cell Carcinoma by Suppression of miR-214-3p Expression
Figure 5
PIM1 was a downstream target of HOMA11-AS and miR-214-3p in the regulation of CDDP chemosensitivity in CDDP-resistant OSCC cells. (a) Bioinformatics analysis indicated the putative binding sites and corresponding mutant region for PIM1 within miR-214-3p. (b) Effect of miR-214-3p transfection on the luciferase activity of WT-PIM1 and MUT-PIM1 reporter systems was evaluated by dual luciferase reporter assay in 293T cells (p=0.029). (c) mRNA expression level of PIM1 in tumor tissues and corresponding noncancerous tissues of 31 OSCC patients (p=0.015). (d) mRNA expression level of PIM1 in OSCC cell lines (TSCCA and CAL-27) and their corresponding CDDP-resistant cell lines (TSCCA-CDDP and CAL-27-CDDP) (p=0.022; p=0.018). ((e) and (f)) TSCCA-CDDP and CAL-27-CDDP cells were transfected with miR-214-3p, or anti- miR-214-3p, or their negative controls. mRNA expression level of PIM1 was determined using real-time PCR ((e) p=0.028; p=0.032; (f) p=0.012; p=0.019). ((g) and (h)) TSCCA-CDDP and CAL-27-CDDP cells were cotransfected with si-HOMA11-AS and anti-miR-214-3p or their negative controls. mRNA expression level of PIM1 was determined using real-time PCR ((g) p=0.031; p=0.036; (h) p=0.037; p=0.041). ((i)–(p)) TSCCA-CDDP and CAL-27-CDDP cells were cotransfected with anti-miR-214-3p and sh-PIM1 or their negative controls. ((i)–(l)) Cell proliferation and IC50 value of CDDP were determined by CCK8 assay kit ((i) p=0.031; p=0.036; (j) p=0.031; p=0.039; (k) p=0.035; p=0.041; (l) p=0.022; p=0.023). ((m) and (n)) Apoptosis was evaluated by TUNEL assay ((m) p=0.021; p=0.027; (n) p=0.022; p=0.031). ((o) and (p)) Caspase 3 activities were detected using assay kit ((o) p=0.025; p=0.033; (p) p=0.026; p=0.032). #P < 0.05.