糖酵解通路关键酶在婴幼儿血管瘤中的表达及其作用  

Expression profiles and roles of key glycolytic enzymes in infantile hemangioma

作　　者：杨开颖 周江元 龚雪 陈思源[2] 吉毅[1] Yang Kaiying;Zhou Jiangyuan;Gong Xue;Chen Siyuan;Ji Yi(Department of Pediatric Surgery,West China Hospital,Sichuan University,Chengdu 610041,China;Department of Critical Care Medicine,West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院小儿外科,成都610041 [2]四川大学华西医院重症医学科,成都610041

出　　处：《中华小儿外科杂志》2022年第11期1007-1012,共6页Chinese Journal of Pediatric Surgery

基　　金：国家自然科学基金资助项目(81401606、81400862);四川大学优秀青年学者基金资助项目(2015SU04A15);四川省科技厅重点研发项目(2019YFS0322);四川大学华西医院学科卓越发展1·3·5工程临床研究孵化项目(2019HXFH056、2020HXFH048、ZYJC21060)。

摘　　要：目的研究糖酵解通路关键酶在婴幼儿血管瘤中的表达变化并探究其作用。方法收集从2020年6月至2020年12月在四川大学华西医院小儿外科行手术切除的8例婴幼儿血管瘤(infan⁃tile hemangioma,IH)组织标本。免疫组织化学分析增殖期组织与消退期组织中糖酵解关键酶的表达,进一步在细胞水平采用Western blot分析在血管瘤内皮细胞(hemangioma⁃derived endothelial cell,HemEC)与人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)的表达情况。应用CCK8、Transwell与Annexin V/PI法检测抑制剂PFK15干预磷酸果糖激酶2/果糖2,6二磷酸酶3(6⁃phosphofructo⁃2⁃kinase/fructose⁃2,6⁃biphosphatase 3,PFKFB3)表达后的HemEC的增殖、迁移及凋亡能力。结果在增殖期血管瘤组织中,糖酵解关键酶葡萄糖转运体1(glucose transporter 1,Glut1)、己糖激酶2(hexokinase 2,HK2)、PFKFB3、磷酸果糖激酶1(phosphofructokinase⁃1,PFK1)、M2型丙酮酸激酶(pyruvate kinase M2,PKM2)及乳酸脱氢酶A(lactate dehydrogenase A,LDHA)的表达水平均高于消退期血管瘤组织。在细胞表达层面,Western blot结果显示,增殖期HemEC中糖酵解关键酶的表达水平均高于HUVEC。使用PFKFB3特异性抑制剂PFK1干预细胞后,HemEC增殖受到显著抑制(P<0.05),HemEC迁移受到显著抑制(163±12比64±7,P<0.01),细胞凋亡升高(11.67±0.74比7.25±0.41,P<0.01)。结论糖酵解关键酶在IH增殖期血管瘤组织中比消退期组织中表达高,在HemEC中表达强于HUVEC。阻断糖酵解关键酶PFKFB3的活性能抑制HemEC增殖,促进凋亡。Objective To explore the expression profiles and roles of key glycolytic enzymes in infantile hemangioma(IH).Methods From June 2020 to December 2020,tissue specimen of 8 hospitalized IH children were collected in West China Hospital.Immunohistochemistry was employed for detecting the expressions of key glycolytic enzymes between proliferating and involuting IH tissues.Then Western blot was utilized for examining the expression profiles between hemangioma⁃derived endothelial cells(HemEC)and human umbilical vein endothelial cell(HUVEC).The roles of PFKFB3 in the proliferation,migration and apoptosis of HemEC were examined after a blockage of PFKFB3.Results In proliferating IH tissues,the expression level of key glycolytic enzymes including Glut1,HK2,PFKFB3,PFK1,PKM2 and LDHA were higher than that in involuting IH tissues.Western blot revealed a higher expression of HemEC as compared with HUVEC.The proliferative activity of HemEC became significantly suppressed after a treatment of PFK15.Furthermore,an inhibition of PFKFB3 with PFK15 could also arrest the migration(163±12 vs 64±7,P<0.01)and accelerate the apoptosis(11.67±0.74 vs 7.25±0.41,P<0.01)of HemEC.Conclusion Key glycolytic enzymes are highly expressed in proliferating IH tissues and HemEC as compared with involuting IH tissues and HUVEC.An inhibition of PFKFB3 may reduce the proliferation,suppress the migration and accelerate the apoptosis of HemEC.And enhanced activities of key glycolytic enzymes may play an important role in the development of IH.

关 键 词：糖酵解 血管瘤 儿童 磷酸果糖激酶2/果糖2 6二磷酸酶3 

分 类 号：R732.2[医药卫生—肿瘤]