Tenascin-C在胶质瘤中的表达及对替莫唑胺疗效的影响

Expression of Tenascin-C in glioma and its effect on temozolomide therapeutic value

目的研究Tenascin-C(TNC)在胶质瘤中的表达特征和临床意义,以及TNC对替莫唑胺(TMZ)治疗胶质瘤疗效的影响。方法通过人类蛋白组数据库和中国胶质瘤患者基因组图谱(CGGA)数据库挖掘TNC在胶质瘤中的表达水平特征及临床意义;通过TNC shRNA质粒和过表达TNC质粒及慢病毒抑制或增强U87MG和U251细胞中TNC的蛋白表达水平,利用CCK-8检测细胞增殖和存活,以及肿瘤细胞U87MG在裸鼠颅内原位成瘤等实验评价TNC对TMZ治疗胶质瘤疗效的影响。结果(1)人类蛋白组数据库显示,TNC在正常神经细胞中不表达或低表达,在脑胶质瘤细胞(U138MG、U87MG、U251)中高转录。(2)CGGA数据库显示,TNC基因在WHOⅣ级胶质母细胞瘤中的转录水平高于WHOⅡ、Ⅲ级胶质瘤(P<0.05)。异柠檬酸脱氢酶(IDH)野生型组TNC mRNA水平显著高于IDH突变型组,无1p/19q联合缺失组显著高于1p/19q联合缺失组(均P<0.05)。高TNC mRNA水平患者的中位生存期(17.33个月)显著短于低TNC mRNA水平患者(89.87个月)(P<0.05)。(3)TNC过表达后显著减弱了TMZ对U87MG和U251细胞增殖的抑制作用和肿瘤杀伤作用(P<0.05),而干扰TNC表达后显著增强了TMZ对U87MG和U251细胞增殖的抑制作用和肿瘤杀伤作用(P<0.05)。干扰TNC表达后显著增强了TMZ对裸鼠颅内U87MG的抑制作用(P<0.05),而过表达TNC后显著减弱了TMZ对裸鼠颅内U87MG的抑制作用(P<0.05)。结论TNC在胶质瘤中高表达,并且与患者的生存预后负相关。TNC水平显著调控TMZ治疗胶质瘤的效果,抑制TNC的表达可能是改善TMZ疗效的有效手段。

Objective To investigate the expression characteristics and clinical relevance of Tenascin-C(TNC)in glioma and the effect of TNC on the efficacy of temozolomide(TMZ)in the treatment of glioma.Methods The Human Protein Atlas and Chinese Glioma Genome Atlas(CGGA)database were used to evaluate the expression level of TNC in glioma and its clinical significance.TNC shRNA plasmid and overexpression plasmid of TNC,and the related lentivirus were used to inhibit or enhance the expression of TNC protein in U87MG and U251 cells.Cell proliferation and survival were detected by CCK-8 assay,and tumor cells U87MG in situ tumor formation in nude mice,were used to evaluate the effect of TNC on the efficacy of TMZ in the treatment of glioma.Results(1)The Human Protein Atlas database showed that TNC was not expressed or low expressed in normal nerve cells,and was highly expressed in glioma cells(U138MG,U87MG,U251).(2)In the CGGA database,the transcription level of TNC in WHO gradeⅣglioblastoma was significantly higher than that in WHO gradeⅡand WHO gradeⅢglioblastoma(P<0.05).The mRNA level of TNC in IDH wild-type glioma was significantly higher than that in IDH mutant glioma (P<0.05), as well as significantly higher in patients without 1P/19q combined deletion glioma than 1P/19q combined deletion glioma (P<0.05). Patients with high TNC mRNA levels had significantly shorter survival outcomes (17.33 months) compared with those with low TNC mRNA levels(89.87 months) (P<0.05). (3)TNC overexpression significantly reduced the proliferation inhibition and tumor killing effect of TMZ on U87MG and U251 cells (P<0.05), and interference with TNC expression significantly enhanced the proliferation inhibition and tumor killing effect of TMZ on U87MG and U251 cells (P<0.05). Interfering TNC expression significantly enhanced the inhibitory effect of TMZ on intracranial U87MG in nude mice (P<0.05), and the overexpression of TNC significantly reduced the inhibitory effect of TMZ on intracranial U87MG in nude mice (P<0.05). Conclusion TNC is highly expressed in glioma and is negatively correlated with survival prognosis of patients. TNC level significantly regulates the efficacy of TMZ in the treatment of glioma, and inhibition of TNC expression may be an effective means to improve the efficacy of TMZ.