BMP-2基因在甲状腺未分化癌细胞中的表达及功能研究

The expression and function of BMP-2 gene in anaplastic thyroid carcinoma

目的:本文主要研究骨形态发生蛋白-2(BMP-2)基因在甲状腺未分化癌细胞中的表达及其功能。方法:通过肿瘤细胞系百科全书(CCLE)和基因表达谱互作分析(GEPIA)数据库比较BMP-2在不同肿瘤细胞中的表达情况并分析其表达差异对甲状腺癌患者生存的影响。进行qRT-PCR验证在甲状腺未分化癌CAL-62细胞中BMP-2 mRNA的表达。再将sh-BMP-2、sh-NC质粒转染入CAL-62细胞,通过CCK-8、克隆形成、流式细胞术、transwell小室试验以及蛋白质印迹实验研究BMP-2基因对CAL-62细胞增殖、凋亡和侵袭的影响。结果:与其他肿瘤细胞相比,BMP-2 mRNA在甲状腺肿瘤细胞中表达最高,且BMP-2高表达患者的总生存时间显著短于低表达组(P=0.010)。CCK-8、克隆形成、流式细胞术以及transwell小室实验表明敲低BMP-2基因能抑制CAL-62细胞的增殖和侵袭并促进其凋亡;蛋白质印迹实验检测到Bcl-2和N-cadherin表达降低而Bax、E-cadherin表达升高。结论:BMP-2高表达与甲状腺肿瘤患者预后不良显著相关;其能促进甲状腺未分化癌细胞的增殖和侵袭并抑制其凋亡。BMP-2基因可能是甲状腺未分化癌潜在的生物标志物和分子治疗靶点。

Objective: In this study,we investigated the expression and function of bone morphogenetic protein-2( BMP-2) in anaplastic thyroid carcinoma. Methods: The expression levels of BMP-2 in various cancer cells were illustrated by CCLE;the association of the expression of BMP-2 and the overall survival of patients with thyroid cancer were analyzed via GEPIA database. We performed qRT-PCR to verify the expression of BMP-2 mRNA in CAL-62 cells. Then,sh-BMP-2 and sh-NC plasmids were transfected into CAL-62 cells. The experiments of CCK-8,clone formation,flow cytometry,transwell and Western blotting were conducted to study the effects of BMP-2 gene on proliferation,apoptosis and invasion of CAL-62 cells. Results: The highest expression level of BMP-2 mRNA was thyroid tumor cells;the overall survival of patients with BMP-2 expression-high was significantly shorter than that of cases with BMP-2 expression-low( P = 0. 010). The CCK-8,clone formation,flow cytometry and transwell assays demonstrated that the knockdown of BMP-2 gene could remarkably inhibit proliferation and invasion but promote apoptosis of CAL-62 cells. Moreover,the Western blotting detected the decreased expressions of Bcl-2 and N-cadherin while the increased expressions of Bax and E-cadherin. Conclusion: Overexpression of BMP-2 is significantly associated with poor prognosis of patients with thyroid tumor;it accelerates proliferation and invasion but impedes apoptosis of undifferentiated thyroid cancer cells. BMP-2 might be a potential biomarker and molecular therapeutic target for anaplastic thyroid cancer.