TGFBI在婴幼儿血管瘤组织中的表达水平及其对婴幼儿血管瘤生物学行为的影响
Expression of TGFBI in infantile hemangioma tissues and its effect on the biological characteristics of hemangioma endothelial cells
作者：李名扬, 杨恩黎, 李易铭, 耿一鸣, 吴海威, 张东升
Author：Li Mingyang, Yang Enli, Li Yiming, Geng Yiming, Wu Haiwei, Zhang Dongsheng
收稿日期：2022-07-04 年卷（期）页码：2023,41(1):29-29-36
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：婴幼儿血管瘤,TGFBI,质粒,增殖,迁移,管腔形成,糖酵解,
Key words：infantile hemangioma,TGFBI,plasmid,proliferation,migration,tube formation,glycolysis,
基金项目：国家自然科学基金(81901022);山东省自然科学基金(ZR2020QH157)
中文摘要
目的 探索TGFBI在不同时期婴幼儿血管瘤组织中的表达水平，并研究质粒转染使TGFBI过表达、小干扰RNA转染使TGFBI敲低对增殖期婴幼儿血管瘤中血管瘤内皮细胞（HemECs）生物学行为的影响。 方法 通过免疫荧光检测TGFBI在不同时期血管瘤组织中的表达水平。构建TGFBI过表达质粒和阴性对照质粒，并分别将其转染至HemECs细胞中；构建TGFBI小干扰RNA及其阴性对照，并将其转染至HemECs细胞中。通过蛋白质印迹（Western blot）检测TGFBI在TGFBI过表达组（OE组）及其阴性对照组（NC组）、TGFBI敲低组（si-TGFBI组）及其阴性对照组（si-NC组）HemECs中的表达水平以验证其转染效果。通过CCK-8试验检测转染后各组细胞的活性，EdU实验检测细胞的增殖率，Transwell检测细胞的迁移能力，管腔形成实验检测细胞的管腔形成能力，细胞外酸化速率（ECAR）试验检测细胞糖酵解水平。 结果 免疫荧光结果显示，TGFBI在增殖期婴幼儿血管瘤组织中的表达高于消退期。Western blot结果显示，OE组TGFBI表达水平高于NC组，si-TGFBI组TGFBI表达水平低于si-NC组。细胞实验中，OE组细胞活力、增殖率、迁移能力及管腔形成能力高于NC组，si-TGFBI组HemECs细胞活性、增殖率、迁移能力及管腔形成能力低于si-NC组。ECAR试验中，OE组糖酵解水平高于NC组。 结论 TGFBI在增殖期血管瘤组织中表达高于消退期。TGFBI的表达上调促进了细胞活性、增殖、迁移和管腔形成能力，且细胞内糖酵解水平上升。TGFBI可能是通过增强糖酵解促进血管瘤发生发展的重要影响因子，是潜在治疗靶点。
英文摘要
ObjectiveThis study aimed to investigate the expression of TGFBI in infantile hemangioma (IH) of proliferative stage or involuting stage and detect the effects of TGFBI overexpression or knockdown on the biological beha-vior of hemangioma endothelial cells (HemECs) from proliferative IH by using plasmid and siRNA.MethodsTGFBI expression levels in proliferative IH and involuting IH were detected by immunofluorescence. TGFBI overexpression plasmid and negative control plasmid were constructed and transfected into HemECs. siRNA for TGFBI and its negative control siRNA were constructed and transfected into HemECs. Western blot was used to detect the expression of TGFBI in the TGFI overexpression group (OE group) and its negative control (NC group), as well as TGFBI knockdown group (si-TGFBI group) and its negative control (si-NC group), to confirm the efficiency of transfection. CCK-8 assays were performed to assess the viability of HemECs. EdU assays were conducted to investigate the proliferation ability of HemECs. Transwell assays were used to detect the migration ability of HemECs. Tube formation assays were carried out to assess the angiogenic capacity of HemECs. Extracellular acidification rate (ECAR) assays were performed to investigate the glycolysis level of HemECs.ResultsThe results of immunofluorescence showed that TGFBI expression was significantly elevated in proliferative IH compared with that in involuting IH. Western blot showed that TGFBI expression in the OE group was upregulated compared with that in the NC group, and TGFBI expression in si-TGFBI was downregulated compared with that in the si-NC group. The viability, cell proliferation, migration ability, and angiogenic capacity of HemECs were promoted in the OE group compared with those in the NC group, whereas these biological behaviors were inhibited in the si-TGFBI group compared with those in the si-NC group. In ECAR assays, the glycolysis level of HemECs in the OE group was enhanced compared with that in the NC group.ConclusionTGFBI is upregulated in proliferative IH. TGFBI overexpression enhanced the viability, cell proliferation, migration ability, and angiogenic capacity of HemECs, which indicated that TGFBI might play a key role in IH progression by accelerating glycolysis. Thus, targeting TGFBI might be an effective therapeutic strategy for IH.
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