Effects of TRIB3 activation of Wnt/β-catenin signaling pathway on growth and proliferation of laryngeal carcinoma TU686 cells in vitro and peripheral immunosuppressive molecule expression in transplanted mice
10.3969/j.issn.1000-484X.2023.12.023
- VernacularTitle:TRIB3激活Wnt/β-catenin信号通路对喉癌TU686细胞体外生长增殖及移植瘤小鼠外周免疫抑制分子表达的影响
- Author:
Zhongqiang CHENG
1
;
Chengyi JIANG
;
Wei WANG
;
Hualong QIANG
;
Xiaodong ZHAN
;
Runsheng YUAN
Author Information
1. 蚌埠医学院第一附属医院耳鼻咽喉头颈外科,蚌埠 233003
- Keywords:
Tribbles homolo 3;
Laryngeal carcinoma;
Proliferation;
Wnt;
β-catenin;
Immunosuppression
- From:
Chinese Journal of Immunology
2023;39(12):2595-2600
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of TRIB3 activation of Wnt/β-catenin signaling pathway on the growth and pro-liferation of laryngeal carcinoma TU686 cells in vitro and expression of peripheral immunosuppressive molecules in transplanted mice.Methods:Protein and RNA expressions of TRIB3 were detected in vitro cells(human immortalized epidermal cell line HaCat and laryngeal carcinoma cell line TU686)and tissues(laryngeal carcinoma and adjacent tissues),respectively.Laryngeal carcinoma TU686 cells were cultured in vitro and divided into negative control group(NC group)and TRIB3 knockdown group(sh-TRIB3 group),total protein and RNA of cells were extracted to verify the expression level of TRIB3 in two groups.After successful verifica-tion,proliferation ability of TU686 cells was detected by CCK-8,colony cloning and flow cytometry.Protein expression levels of Wnt,Cyclin-D1,C-myc,β-catenin and p-β-catenin in two groups were detected by Western blot.Correlation analysis verified the correla-tion between TRIB3 and Wnt,Cyclin-D1,C-myc,β-catenin,p-β-catenin protein expressions.TRIB3-low expressing nude mouse transplanted tumor model(TRIB3 sgRNA group)was constructed by knockdown the TRIB3 core plasmid,and a parallel control group(Control sgRNA group)was set up,tumor growth volume and weight were observed,and serum immunosuppressive molecules expres-sions was determined by ELISA.Results:Compared with HaCat cells and normal paracarcinoma tissues,TRIB3 was highly expressed in TU686 cells and laryngeal carcinoma tissues.Compared with negative control group,proliferation ability of TU686 cells was signifi-cantly inhibited after TRIB3 knockdown,and cell growth was blocked in G1/S phase.Expressions of Wnt,Cyclin-D1,C-myc andβ-catenin protein in Wnt/β-catenin signaling pathway were decreased significantly,while expression of p-β-catenin was increased significantly.TRIB3 was significantly correlated with protein expression levels of Wnt,Cyclin-D1,β-catenin and p-β-catenin.The in vivo results showed that compared with Control sgRNA group,tumor growth volume and weight of mice in TRIB3 sgRNA group were significantly decreased,and expressions of serum immunosuppressive molecules IL-4,IL-6,IL-10,TGF-β and PGE2 were signifi-cantly decreased.Conclusion:TRIB3 is highly expressed in TU686 cells,and TRIB3 can inhibit growth and proliferation of TU686 cells and transplanted tumors by activating Wnt/β-catenin-related signaling pathways,and reverse tumor immunosuppressive microen-vironment,suggesting that TRIB3 may be an effective target for laryngeal cancer.
