IDH突变促进胶质瘤血管生成拟态形成的研究

A Study on IDH Mutation Promoting the Formation of Angiogenic Mimicry in Glioma

目的:观察IDH1突变胶质瘤血管生成拟态的形成特点。方法:实验分为:突变IDH表达组(dox+组),野生IDH表达组(dox−组)。Western blot检测dox+组胶质瘤U87细胞内突变IDH1蛋白表达,HE染色观察两组细胞形态变化;管形成实验检测血管生成拟态形成;6周龄裸鼠皮下注射建立移植瘤模型,观察胶质瘤裸鼠移植瘤中的血管生成拟态,通过HE染色、免疫组化检测mutIDH1、CD34表达,高碘酸雪夫(PAS)染色观察基底膜样物形成。结果:胶质瘤U87细胞形态饱满,血管生成拟态显示,dox+组细胞管形成数量明显多于对照组(*P < 0.05);dox+组胶质瘤U87细胞内表达mutIDH1蛋白。成功构建IDH1突变(mutIDH1)人胶质瘤U87细胞移植瘤模型。HE染色显示瘤细胞生长密集,可见多量血管增生。免疫组化显示,dox+组mutIDH1表达明显,dox−组不表达mutIDH1蛋白;CD34的表达在dox−组明显多于dox+组;PAS染色发现,dox+组较dox−有更多血管生成拟态形成。结论:IDH1突变促进血管生成拟态形成。

Objective: To observe the formation characteristics of angiogenesis mimicry in IDH1-mutant glioma. Methods: The experimental groups were divided into IDH mutant group (dox+ group) and IDH non-mutant group (dox− group). Western blot was used to detect the expression of mutIDH1 pro-tein in glioma U87 cells in the dox+ group, and the morphological changes of the two groups were observed by HE staining. The ability to form the tube was tested by the angiogenesis mimic experi-ment. 6-week-old nude mice were injected subcutaneously to establish a xenograft model, and tis-sue experiments were carried out to detect the expression of mutIDH1 and CD34 by HE staining, periodic acid sherf (PAS) reagent staining, and immunohistochemistry. Angiogenesis mimicry for-mation in glioma nude mouse xenografts was observed. Results: The morphology of glioma U87 cells was full, and the angiogenesis mimicry showed that the number of the tubes like structure formed in the dox+ group was significantly more than dox− group (*P < 0.05). Glioma U87 in the dox+ group expressed mutIDH1 protein in cells. An in vitro xenograft tumor model of IDH1 mutant (mutIDH1) human glioma U87 cells was successfully constructed. HE staining showed dense growth of tumor cells and multiple angiogenesis. Immunohistochemistry showed that mutIDH1 was signif-icantly expressed in the dox+ group, but mutIDH1 protein was not expressed in the dox− group. The expression of CD34 was significantly higher in the dox− group than in the dox+ group. PAS staining showed that the dox+ group formed more angiogenic mimics than dox−. Conclusion: IDH1 mutation promotes angiogenesis mimicry.