Signal transducers and activators of transcription-3 modulates human squamous cell carcinoma invasion via targeting mircoRNA-21 in vitro
10.3760/cma.j.issn.1002-0098.2013.09.006
- VernacularTitle:信号传导及转录激活因子调控微RNA-21影响人舌鳞状细胞癌细胞侵袭能力的体外研究
- Author:
Ai-Qin LIU
1
;
Sha-Sha LI
;
Xuan ZHOU
;
Xu-Dong WANG
;
Lun ZHANG
Author Information
1. 300060,天津医科大学附属肿瘤医院颌面耳鼻喉肿瘤科·天津市肿瘤防治重点实验室
- Keywords:
MicroRNA;
Carcinoma,squamous cell;
Neoplasm invasiveness
- From:
Chinese Journal of Stomatology
2013;48(9):539-544
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect and mechanism of signal transducers and activators of transcription 3 (STAT-3) modulates human tongue squamous cell carcinoma invasion ability via targeting mircoRNA-21.Methods Tscca and Tca8113P160 human tongue squamous cell carcinoma cell lines were used.WP1066(STAT-3 inhibitor),the small molecule inhibitor of STAT-3 was used to suppress the STAT-3 expression.The half maximal inhibitory concentration (ICs0 value) of WP1066 in the two cell lines was determined by methyl thiazolyl tetrazolium(MTT) assay.The expression level of STAT-3 and phosphorylation of STAT-3 (pSTAT-3) was examined by Western blotting.Real-time PCR was used to detect the mircoRNA-21 expression after treated with WP1066.Matrigel matrix and transwell assay were used to determine cancer cell colony formation and invasion ability after treated with WP1066.Tumor invasion related proteins in Tscca and Tca8113P160 cell lines were measured by Western blotting.Luciferase reporter gene assay was conducted to detect the relationship between STAT-3 and mircoRNA-21.Results The IC50 to WP1066 in Tscca cell was 3.1 and 3.5 μmol/L for Tca8113P160 cell respectively.STAT-3/pSTAT-3 protein level was suppressed significantly(Tscca:STAT-3:F =887.154,P =0.000; pSTAT-3:F =332.212,P =0.000 ; Tca8113P160:STAT-3:F =322.895,P =0.000 ; pSTAT-3:F =788.357,P =0.000).mircoRNA-21 expression was downregulated(Tscca:F =32.157,P =0.000 ; Tca8113P160:F =11.349,P =0.007).The diameters of culture clone in cell treated with WP1066 were less than control groups (Tscca:F =15.751,P =0.004;Tca8113P160:F =12.964,P =0.007).The number of tongue cancer cell migrating through the transwell membrane in WP1066 treated group was less than in control groups (Tscca:F =1688.926,P =0.000;Tca8113P160:F =327.528,P =0.000).In addition,MMP-2/9 protein expression was decreased in both of the cell lines treated with WP1066,while TIMP-3 was up regulated dramatically.STAT-3 could modulate mircoRNA-21 directly.Conclusions Reduction of STAT-3 can inhibit tongue cancer cell invasion ability via targeting mircoRNA-21.
