摘要目的 探讨Twist2在神经胶质瘤中的表达意义及其是否通过上皮间质转化参与胶质瘤的恶性转化.方法 采用免疫组织化学法检测60例脑胶质瘤(包括WHO Ⅱ级、Ⅲ级、Ⅳ级各20例)及20例非肿瘤脑组织中Twist2蛋白的表达水平;采用即时荧光定量PCR法和Western blot分别检测61例胶质瘤(WHO Ⅱ级16例、Ⅲ级21例、Ⅳ级24例)和12例瘤旁新鲜组织中Twist2 mRNA和蛋白的表达水平,同时应用Western blot分别检测各组新鲜组织标本中E-cadherin、N-cadherin、波形蛋白表达水平.结果 免疫组织化学法结果显示:Twist2在胶质瘤中的表达阳性率为90%(54/60),在非肿瘤脑组织中为30%(6/20),两者差异有统计学意义(P<0.01);在胶质瘤WHO Ⅱ级、Ⅲ级、Ⅳ级中的表达阳性率分别为75%(15/20)、95%(19/20)、100%(20/20).WHO Ⅳ级、Ⅲ级显著高于Ⅱ级(P<0.01),WHOⅣ级与Ⅲ级的表达,差异无统计学意义(P>0.05).即时荧光定量PCR法和Western blot结果显示:Twist2在胶质瘤中的表达水平明显高于瘤旁组织(P<0.01);且WHOⅣ级、Ⅲ级中的表达均明显高于Ⅱ级,差异有统计学意义(P<0.01);胶质瘤WHOⅣ级和Ⅲ级比较,差异无统计学意义(P>0.05);Western blot检测上皮间质转化中关键蛋白表达水平结果:新鲜胶质瘤组织中E-cadherin蛋白表达水平与Twist2呈负相关(r=-0.972,P<0.01);N-cadherin、波形蛋白表达水平与Twist2均呈正相关(r=0.971,P<0.01;r=0.968,P<0.01).结论 Twist2在人神经胶质细胞瘤中存在表达,且与胶质瘤恶性等级呈正相关,其可能通过上皮间质转化参与胶质瘤的恶性进程.

abstractsObjective To investigate the significance of Twist2 in glioma and whether it is involved in the malignant transformation of glioma by epithelial-mesenchymal transition(EMT). Methods Using immunohistochemical method detected the expression level of Twist2 in 60 cases of gliomas(including WHO grades Ⅱ,Ⅲ and Ⅳ,each for 20 cases)and 20 cases of non-tumor brain tissues. Real-time fluorescence quantitative PCR and Western blot were used to detect the expression level of Twist2 mRNA and protein in 61 cases of fresh glioma tissue(WHO grade Ⅱ 16 cases, Ⅲ 21 cases, Ⅳ 24 cases)and 12 cases of adjacent tissues,and the expression levels of E-cadherin,N-cadherin and vimentin were also investigated in fresh glioma tissue. Results Immunohistochemistry Results showed that the percentages of Twist2 expression in glioma was 90%(54/60)compared with 30%(6/20)in non-tumor brain tissues(P<0.01). The percentages of Twist2 expression were 75%(15/20), 95%(19/20), and 100%(20/20)in the WHO gradesⅡ,Ⅲ and Ⅳ gliomas, respectively. WHO grades Ⅳ and Ⅲ were significantly higher than that of WHO grade Ⅱ(P<0.01). There was no significant difference between WHO gradeⅣand WHOⅢglioma (P>0.05). Real-time fluorescence quantitative PCR and Western blot showed that the expression level of Twist 2 in gliomas was significantly higher than that in para-cancerous tissues(P<0.01), and those in WHO grades Ⅳ and Ⅲ gliomas were significantly higher than that in WHO grade Ⅱ glioma(P<0.01). There was no significant difference between WHO gradeⅣand gradeⅢglioma(P>0.05). Detection of key protein expression in EMT by Western blot displayed that the expression of E-cadherin was negatively associated with Twist2 in glioma(r=-0.972,P<0.01). The expression of N-cadherin and vimentin was positively associated with Twist2 in glioma(r=0.971,P<0.01;r=0.968,P<0.01). Conclusions The expression of Twist2 in human glioma is positively correlated with the malignant grade of glioma,which may be involved in the malignant progression of glioma by EMT.