2型转谷氨酰胺酶对人胶质母细胞瘤细胞U251放射敏感性的影响

辐射防护 ›› 2024, Vol. 44 ›› Issue (5): 537-545.

2型转谷氨酰胺酶对人胶质母细胞瘤细胞U251放射敏感性的影响

蓝瑞隆, 伍兵, 王锃

福建医科大学附属第一医院中心实验室,放射生物福建省高等学校重点实验室,福建省肿瘤精准诊疗重点实验室,福建医科大学附属第一医院滨海院区国家区域医疗中心,福州 350005

收稿日期:2023-08-07 出版日期:2024-09-20 发布日期:2024-11-05
通讯作者: 王锃。E-mail: wzishere@126.com
作者简介:蓝瑞隆(1986—),男,2009年本科毕业于中央民族大学生物科学专业,2012年硕士毕业于中国医学科学院&北京协和医学院生物医学工程专业,助理研究员。E-mail: lanruilong123@126.com
基金资助:
福建省自然科学基金项目(2022J01676);福建省科技创新联合资金项目(2021Y9110);福建省卫生健康中青年骨干人才培养项目(2021GGA027)。

Effects of transglutaminase 2 on radiosensitivity of human glioblastoma cell line U251

LAN Ruilong, WU Bing, WANG Zeng

Central Lab of The First Affiliated Hospital of Fujian Medical University; Key Laboratory of Radiation Biology of Fujian Province; Fujian Provincial Key Laboratory of Precision Medicine for Cancer Universities; National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou 350005

Received:2023-08-07 Online:2024-09-20 Published:2024-11-05

摘要/Abstract

摘要： 为探讨2型转谷氨酰胺酶(transglutaminase 2,TG2)对胶质母细胞瘤(glioblastoma,GBM)放射敏感性的影响。通过生信分析TG2表达与胶质瘤病人预后的关系,并分析了其在GBM放射抵抗细胞株U251中的水平;瞬时转染得到TG2过表达细胞(U251-TG2)或空载细胞(U251-EV);CCK8法绘制生长曲线;transwell检测细胞迁移能力;不同剂量X射线照射后,检测细胞克隆形成能力、ROS水平;并通过免疫荧光检测γ-H2AX水平;Western blot检测自噬关键分子Beclin-1、LC3及mTOR磷酸化水平。结果显示,TG2高表达胶质瘤患者具有更差的无进展间隔期及总生存期,TG2在放射抵抗型U251中的表达显著高于野生型U251;U251-TG2细胞增殖能力高于U251-EV,且前者迁移能力高于后者;经X射线照射后,U251-TG2细胞克隆形成率高于U251-EV,前者ROS水平及γ-H2AX荧光焦点数均低于后者;自噬关键蛋白Beclin-1及自噬活性指标LC3-Ⅱ/Ⅰ比值在U251-TG2的水平均高于U251-EV,且前者mTOR磷酸化水平低于后者。实验结果证明,X射线照射后,TG2可能通过抑制mTOR磷酸化的方式促进自噬,进而降低GBM细胞U251的放射敏感性。

关键词: 胶质母细胞瘤, 2型转谷氨酰胺酶, 放射敏感性

Abstract: To investigate the effect of transglutaminase 2 (TG2) on radiosensitivity of glioblastoma, bioinformatic analysis was adopted to analyze the relationship between TG2 expression and the prognosis of glioma patients and TG2 level in GBM radioresistant cell line U251; TG2-overexpression cells (U251-TG2) or control cells (U251-EV) were obtained by transient transfection; cell growth curve was drawn by CCK8 method, and cell migration ability was measured by transwell; colony formation ability, ROS level and γ-H2AX foci were measured after X-ray treatment; Beclin-1, LC3 and mTOR phosphorylation levels were detected by Western blot. The results showed that glioma patients with high TG2 expression had worse progression-free interval and overall survival, and TG2 expression in radioresistant U251 was significantly higher than that in wild type U251; the cell proliferation and migration ability of U251-TG2 were higher than that of U251-EV; after X-ray treatment, compared with U251-EV, U251-TG2 showed higher colony-forming ratio, lower ROS level and γ-H2AX foci number; the levels of Beclin-1 and LC3-II/I in U251-TG2 were higher than those in U251-EV; mTOR phosphorylation level in U251-TG2 was lower than that in U251-EV. The results indicated that TG2 could promote autophagy by inhibiting mTOR phosphorylation, and then decrease the radiosensitivity of U251 cells.

Key words: glioblastoma, transglutaminase 2, radiosensitivity

中图分类号:

Q274

蓝瑞隆, 伍兵, 王锃. 2型转谷氨酰胺酶对人胶质母细胞瘤细胞U251放射敏感性的影响[J]. 辐射防护, 2024, 44(5): 537-545.

LAN Ruilong, WU Bing, WANG Zeng. Effects of transglutaminase 2 on radiosensitivity of human glioblastoma cell line U251[J]. RADIATION PROTECTION, 2024, 44(5): 537-545.

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