Β3-微管蛋白Ⅲ类在儿童髓母细胞瘤中的表达及其机制研究

Expression and mechanism of β-tubulin isotype Ⅲ in children with medulloblastoma

摘要:

目的 探讨β3-微管蛋白Ⅲ类（TUBB3）在儿童髓母细胞瘤中的表达及其机制。方法 免疫组织化学检测TUBB3、程序性死亡受体配体1（PD-L1）及肿瘤干细胞标志物在儿童髓母细胞瘤中的表达，并分析其相关性。CCK-8及Transwell法检测TUBB3对髓母细胞瘤细胞增殖和侵袭的影响。免疫印迹法（Western blotting）分析P53对TUBB3表达的影响。结果 TUBB3阳性表达率为82.1%。TUBB3表达与组织分化程度、淋巴结转移及雌激素受体水平有关。Kaplan-Meier分析显示，TUBB3高表达与髓母细胞瘤术后总生存率低有关。敲低TUBB3表达能抑制Daoy和D341细胞增殖侵袭。儿童髓母细胞瘤中TUBB3与CD44、CD133、PD-L1及P53表达呈正相关。免疫印迹法显示，p53基因敲低使TUBB3表达上调。结论 TUBB3在儿童髓母细胞瘤中发挥致癌作用，是儿童髓母细胞瘤的潜在生物标志物和治疗靶点。

Abstract:

Objective To investigate the expression and mechanism of β-tubulin isotype Ⅲ （TUBB3） in children with medulloblastoma.Methods Immunohistochemistry was used to measure the expression of TUBB3， programmed death-ligand 1 （PD-L1）， and tumor stem cell markers in children with medulloblastoma， and their correlation was analyzed. CCK-8 assay and Transwell assay were used to observe the effect of TUBB3 on the proliferation and invasion of medulloblastoma cells， and Western blotting was used to analyze the influence of P53 expression on TUBB3 expression.Results The positive rate of TUBB3 was 82.1%. The expression of TUBB3 was associated with the degree of tissue differentiation， lymph node metastasis， and the level of estrogen receptor. A Kaplan-Meier analysis showed that the high expression of TUBB3 was associated with the low overall survival rate of medulloblastoma after surgery. Knockdown of TUBB3 inhibited the proliferation and invasion of Daoy and D341 cells. TUBB3 was positively correlated with the expression of CD44， CD133， PD-L1， and P53 in children with medulloblastoma. Western blotting showed that the expression of TUBB3 was upregulated by p53 gene knockdown.Conclusion TUBB3 plays a carcinogenic role in children with medulloblastoma and is a potential biomarker and therapeutic target for medulloblastoma in children.