ATF3、14-3-3ζ和beclin-1在替莫唑胺治疗敏感与耐药的胶质母细胞瘤中的表达及意义
| 作者: |
1李小燕,
2赵建祥,
3任保刚,
1吴雪晶,
1吴丹,
1姚梅宏,
1陈晓雯,
1杨映红
1 福建医科大学附属协和医院病理科，福州 350001 2 福建省立医院重症医学三科，福州 350001 3 福建医科大学附属协和医院神经外科，福州 350001 |
| 通讯: |
杨映红
Email: yyh1555@163.com |
| DOI: | 10.3978/j.issn.2095-6959.2021.12.004 |
| 基金: | 福建省科技创新联合资金项目(2017Y9034)；福建省卫生健康科技计划青年项目(2016-1-50)；福建医科大学启航基金项目(2016QH021)。 |
摘要
目的：探讨在替莫唑胺(temozolomide，TMZ)治疗敏感与耐药的胶质母细胞瘤中转录激活因子3(activating transcription factor 3，ATF3)、14-3-3ζ和beclin-1的表达水平变化及其与TMZ耐药产生的相关意义。方法：采用RT-PCR及免疫组织化学法检测并比较同一患者TMZ治疗前及耐药时胶质母细胞瘤组织内ATF3、14-3-3ζ和beclin-1基因及蛋白质表达水平。取30 μg/mL TMZ培养的U87细胞为实验组，普通溶剂培养的U87细胞为对照组，培养72 h，MTT及annexin V-别藻蓝素(allophycocyanin，APC)单染法分别检测并比较肿瘤细胞的生长抑制率、凋亡率，RT-PCR及蛋白质印迹检测并比较细胞内ATF3、14-3-3ζ和beclin-1基因及蛋白质表达水平。结果：TMZ耐药时胶质母细胞瘤组织中ATF3、14-3-3ζ和beclin-1表达水平均高于TMZ治疗前(P<0.05)；TMZ敏感的U87细胞培养72 h肿瘤细胞生长抑制率和凋亡率均增加(P<0.05)，细胞内ATF3、14-3-3ζ和beclin-1表达水平均低于对照组(P<0.05)；三者呈同向表达。结论：在TMZ用药过程中，胶质母细胞瘤内ATF3激活可能上调14-3-3ζ的表达，进而启动并使自噬增强，导致TMZ耐药产生。
关键词：
胶质母细胞瘤；替莫唑胺；自噬；转录激活因子3；14-3-3ζ
Expression of ATF3, 14-3-3ζ, and beclin-1 in the treatment of sensitive and drug-resistant glioblastomas with temozolomide and its significance
CorrespondingAuthor: YANG Yinghong Email: yyh1555@163.com
DOI: 10.3978/j.issn.2095-6959.2021.12.004
Foundation: This work was supported by the Joint Funds for the Innovation of Science and Technology, Fujian Province (2017Y9034), Youth Research Projects sponsored by Fujian Provincial Health Technology Project (2016-1-50), and Fujian Medical University Start-up Fund Project (2016QH021), China.
Abstract
Objective: To investigate the changes of the expression of activating transcription factor 3 (ATF3), 14-3-3ζ, and beclin-1 in the treatment of sensitive and drug-resistant glioblastomas with temozolomide (TMZ). And to investigate their correlation with TMZ resistance. Methods: Before TMZ treatment and after TMZ resistance, the gene and protein expression levels of ATF3, 14-3-3ζ, and beclin-1 in glioblastoma tissues of the same patient were detected and compared by RT-PCR and immunohistochemistry. U87 cells cultured with 30 μg/mL TMZ were taken as an experimental group, and U87 cells cultured with common solvent were taken as a control group. After 72 h of culture, the growth inhibition rate and apoptotic rate of tumor cells between the two groups were detected and compared by MTT and annexin V-allophycocyanin (APC) monochrome staining, respectively. After 72 h of culture, the gene and protein expression levels of ATF3, 14-3-3ζ, and beclin-1 between the two groups were detected and compared by RT-PCR and Western blotting. Results: The expression of ATF3, 14-3-3ζ and beclin-1 in glioblastoma tissues with TMZ resistance was higher than that before TMZ treatment (P<0.05). After 72 h of culture, the growth inhibition rate and apoptotic rate of TMZ-sensitive U87 cells were increased (P<0.05); meanwhile, the expression of ATF3, 14-3-3ζ and beclin-1 in TMZ-sensitive U87 cells was lower than that in the control group (P<0.05). All three have the same expression trend. Conclusion: During the TMZ treatment process, the activation of ATF3 in glioblastoma may upregulate the expression of 14-3-3ζ which initiates and enhances the autophagy of tumor cells, leading to TMZ resistance in glioblastoma.
Keywords：
glioblastoma; temozolomide; autophagy; activating transcription factor 3; 14-3-3ζ