School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University, Jinan 250012, China.
Dept. of Stomatology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan 250021, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2023 Feb 1;41(1):29-36. doi: 10.7518/hxkq.2023.01.004.
OBJECTIVES: This 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. METHODS: TGFBI 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. RESULTS: The 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. CONCLUSIONS: TGFBI 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.
目的：本研究旨在探讨转化生长因子β诱导（TGFBI）在婴儿血管瘤（IH）增殖期或消退期的表达，并通过质粒和 siRNA 检测 TGFBI 过表达或敲低对增殖期 IH 血管内皮细胞（HemECs）生物学行为的影响。
方法：采用免疫荧光法检测增殖期 IH 和消退期 IH 中 TGFBI 的表达水平。构建 TGFBI 过表达质粒和阴性对照质粒，并转染至 HemECs。构建 TGFBI siRNA 及其阴性对照 siRNA，并转染至 HemECs。Western blot 检测 TGFBI 过表达组（OE 组）及其阴性对照（NC 组）、TGFBI 敲低组（si-TGFBI 组）及其阴性对照（si-NC 组）中 TGFBI 的表达，以确认转染效率。CCK-8 法检测 HemECs 活力。EdU 法检测 HemECs 增殖能力。Transwell 法检测 HemECs 迁移能力。管腔形成实验评估 HemECs 的血管生成能力。细胞外酸化率（ECAR）实验检测 HemECs 的糖酵解水平。
结果：免疫荧光结果显示，与消退期 IH 相比，增殖期 IH 中 TGFBI 的表达显著上调。Western blot 结果显示，OE 组 TGFBI 表达上调，NC 组 TGFBI 表达下调，si-TGFBI 组 TGFBI 表达下调，si-NC 组 TGFBI 表达下调。与 NC 组相比，OE 组 HemECs 的活力、细胞增殖、迁移能力和血管生成能力均增强，而 si-TGFBI 组 HemECs 的这些生物学行为则受到抑制。在 ECAR 实验中，与 NC 组相比，OE 组 HemECs 的糖酵解水平增强。
结论：TGFBI 在增殖期 IH 中上调。TGFBI 过表达增强了 HemECs 的活力、细胞增殖、迁移能力和血管生成能力，表明 TGFBI 通过加速糖酵解可能在 IH 进展中发挥关键作用。因此，靶向 TGFBI 可能是治疗 IH 的一种有效策略。
Hua Xi Kou Qiang Yi Xue Za Zhi. 2023-2-1
Arterioscler Thromb Vasc Biol. 2023-5
Cell Signal. 2021-8
Expert Rev Clin Pharmacol. 2020-8
Eur J Pediatr. 2019-11-22
Lancet. 2017-1-13
Adv Neonatal Care. 2016-4
Front Surg. 2014-9-25
Zotero 插件