中华皮肤科杂志 ›› 2023, Vol. 56 ›› Issue (4): 320-324.doi: 10.35541/cjd.20220463

• 论著 • 上一篇    下一篇

糖酵解关键酶PFKFB3对婴幼儿血管瘤内皮细胞增殖、迁移及凋亡的影响

杨开颖    龚雪    邱桐    周江元    兰钰茹    吉毅   

  1. 四川大学华西医院小儿外科，成都  610041
  • 收稿日期:2022-06-24 修回日期:2023-01-11 发布日期:2023-03-31
  • 通讯作者: 吉毅 E-mail:jijiyuanyuan@163.com
  • 基金资助:
    国家自然科学基金项目（82273556）；四川省科技厅重点研发项目（2022YFS0233、2022YFS0225、2022NSFSC1480）；四川大学从0到1创新研究项目（2022SCUH0033）；四川大学医学+信息中心交叉学科建设开放项目（YGJC004）；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（ZYJC21060、2020HXFH048、2019HXFH056）

Effects of the key glycolytic enzyme PFKFB3 on the proliferation, migration and apoptosis of hemangioma-derived endothelial cells

Yang Kaiying, Gong Xue, Qiu Tong, Zhou Jiangyuan, Lan Yuru, Ji Yi   

  1. Department of Pediatric Surgery, West China Hospital, Sichuan University, Chengdu 610041, China
  • Received:2022-06-24 Revised:2023-01-11 Published:2023-03-31
  • Contact: Ji Yi E-mail:jijiyuanyuan@163.com
  • Supported by:
    National Natural Science Foundation of China （82273556）; Key Project in the Science & Technology Program of Sichuan Province （2022YFS0233, 2022YFS0225, 2022NSFSC1480）; Project of ‘0 to 1’ of Sichuan University （2022SCUH0033）; Med-X Center for Informatics Funding Project （YGJC004）; The 1·3·5 Project for Disciplines of Excellence Clinical Research Incubation Project, West China Hospital of Sichuan University （ZYJC21060, 2020HXFH048, 2019HXFH056）

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摘要： 【摘要】 目的 研究糖酵解关键酶6-磷酸果糖-2-激酶/果糖-2，6-二磷酸酶3（PFKFB3）对婴幼儿血管瘤（IH）内皮细胞（HemEC）生物活性的影响。方法 收集4例增殖期及4例消退期IH组织，从增殖期IH组织中提取原代HemEC，以脐静脉内皮细胞（HUVEC）为对照。通过免疫组化与Western印迹法分别检测PFKFB3在IH组织与HemEC中的表达水平。采用CCK8实验检测0 ~ 10 μmol/L PFKFB3特异性抑制剂PFK15对HemEC细胞增殖的影响。体外培养HemEC，采用5 μmol/L PFK15干预细胞，采用Transwell法观察HemEC迁移能力，流式细胞仪检测HemEC凋亡水平。组间比较采用t检验或方差分析。结果 免疫组化显示，增殖期IH组织中PFKFB3表达丰度（74.34% ± 5.26%）高于消退期（41.46% ± 2.99%，t = 9.40，P < 0.001）。Western印迹显示，HemEC中PFKFB3相对表达水平（0.73 ± 0.05）高于HUVEC（0.45 ± 0.04，t = 8.50，P < 0.001）。CCK8实验结果显示，0.625、1.25、2.5、5、10 μmol/L PFK15组HemEC增殖活性均低于对照组（均P < 0.01）。Transwell实验显示，PFK15干预组HemEC迁移数（297 ± 15）低于对照组（422 ± 8，t = 12.59，P < 0.001）。流式细胞仪检测显示，PFK15干预组HemEC凋亡率（6.69% ± 0.64%）高于对照组（0.34% ± 0.07%，t = 17.07，P < 0.001）。结论 糖酵解关键酶PFKFB3在IH增殖期组织与HemEC中高表达，PFKFB3抑制剂PFK15可抑制HemEC增殖、迁移并促进其凋亡。

关键词: 血管瘤, 血管瘤内皮细胞, 糖酵解, 细胞增殖, 细胞凋亡, 细胞迁移, 6磷酸果糖2激酶/果糖-2, 6-二磷酸酶3

Abstract: 【Abstract】 Objective To investigate the effect of the key glycolysis enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 （PFKFB3） on the biological activity of hemangioma-derived endothelial cells （HemECs）. Methods Totally, 4 proliferating infantile hemangioma （IH） tissues and 4 involuting IH tissues were collected. Primary HemECs were isolated from the proliferating IH tissues, and human umbilical vein endothelial cells （HUVECs） served as controls. Immunohistochemical study and Western blot analysis were performed to determine the expression of PFKFB3 in the IH tissues and HemECs, respectively. Cell counting kit-8 （CCK8） assay was conducted to evaluate the effect of PFK15 （a specific inhibitor of PFKFB3） at concentrations of 0 - 10 μmol/L on the proliferation of HemECs, and HemECs treated without PFKFB3 served as the control group. Some in vitro cultured HemECs were treated with 5 μmol/L PFK15, and served as a PFK15 intervention group, while HemECs treated without PFK15 served as a control group; then, the migratory ability of HemECs was assessed by Transwell assay, and the apoptosis level of HemECs was detected by flow cytometry. Comparisons between groups were performed by using t test or analysis of variance. Results Immunohistochemical study showed that the positive rate of PFKFB3 was significantly higher in the proliferating IH tissues （74.34% ± 5.26%） than in the involuting IH tissues （41.46% ± 2.99%, t = 9.40, P < 0.001）. Western blot analysis showed that the relative expression level of PFKFB3 was also significantly higher in HemECs （0.73 ± 0.05） than in HUVECs （0.45 ± 0.04, t = 8.50, P < 0.001）. CCK8 assay revealed significantly decreased proliferative activity of HemECs in the 0.625-, 1.25-, 2.5-, 5-, and 10-μmol/L PFK15 groups compared with the control group （all P < 0.01）. Compared with the control group, the PFK15 intervention group showed significantly decreased number of migratory HemECs （297 ± 15 vs. 422 ± 8, t = 12.59, P < 0.001）, but significantly increased apoptosis rates of HemECs （6.69% ± 0.64% vs. 0.34% ± 0.07%, t = 17.07, P < 0.001）. Conclusion The key glycolytic enzyme PFKFB3 was highly expressed in the proliferating IH tissues and HemECs, and the PFKFB3 inhibitor PFK15 could suppress the proliferation, migration, and increase the apoptosis of HemECs.

Key words: Hemangioma, Hemangioma-derived endothelial cell, Glycolysis, Cell proliferation, Apoptosis, Cell migration, 6-Phosphofructo-2-kinase/fructose-2,6-biphosphatase 3

引用本文

杨开颖 龚雪 邱桐 周江元 兰钰茹 吉毅. 糖酵解关键酶PFKFB3对婴幼儿血管瘤内皮细胞增殖、迁移及凋亡的影响[J]. 中华皮肤科杂志, 2023,56(4):320-324. doi:10.35541/cjd.20220463

Yang Kaiying, Gong Xue, Qiu Tong, Zhou Jiangyuan, Lan Yuru, Ji Yi. Effects of the key glycolytic enzyme PFKFB3 on the proliferation, migration and apoptosis of hemangioma-derived endothelial cells[J]. Chinese Journal of Dermatology, 2023, 56(4): 320-324.doi:10.35541/cjd.20220463