目的 探讨不同病理级别国人脑胶质瘤中miR-21表达;miR-21编码基因扩增与突变;miR-21与EGFR信号通路关键成员表达关系.方法 荧光实时定量PCR法和荧光原位杂交法测定不同病理级别人脑胶质瘤标本中miR-21表达;半定量PCR法检测miR-21编码基因拷贝数并测序PCR产物;免疫组织化学染色法检测胶质瘤标本EGFR、pAKT和PTEN表达水平.结果 荧光实时定量PCR与荧光原位杂交miR-21表达随着胶质瘤病理级别的升高而增加(F=35.516;P=0.000);不同病理级别中人脑胶质瘤中miR-21编码基因拷贝数差异无统计学意义(F=1.219;P=0.305);miR-21表达与EGFR(rs=0.710,P=0.000)、pAKT(rs=0.638,P=0.000)表达呈正相关,与PTEN表达呈负相关(rs=-0.286,P=0.027).结论 miR-21在国人脑胶质瘤中过表达且与EGFR信号通路关键成员表达相关.
Objective To detect miR -21 over-expression status in Chinese glioblastoma and the amplification and mutation frequency of miR -21 encoding gene and the correlation between miR -21 over -expression and EGFR signaling transduction pathway. Method The qPCR and fluorescence in - situ hybridization method was conducted to examine miR - 21 relative and in - situ expression across different WHO classified Chinese human gliomas. RT- PCR was used to examine miR -21 encoding gene amplification copy number difference across different grades of GBM and the PCR product was further sequencing analyzed. Immuno - histochemistry staining was used to detect EGFR, pAKT and PTEN expression in GBM. Results miR -21 was statistical different expressed across the WHO classification of gliomas by qPCR and fluorescence in- situ hybridization method( F =35. 516;P =0. 000). There was no difference of miR -21 encoding gene copy number changes in miR -21 differently expressed gliomas and no sense spot mutation of miR -21 encoding gene. The miR -21 expression was positively correlated to those of EGFR( rs =0. 710, P =0. 000) and pAKT( rs =0. 638, P =0. 000) and negatively correlated to PTEN( rs= -0. 286, P = 0. 027) expression. Conclusion miR -21 is abnormal activated in WHO classified Chniese gliomas and related to EGFR signaling pathway.