转录因子MYB转录调控MTFR2通过DNA损伤修复促进胃癌细胞化疗耐药性  

The transcription factor MYB transcriptionally regulates MTFR2 to promote chemotherapy resistance of gastric cancer cells through DNA damage repair

作　　者：李春兴[1] 迪力旦·纳斯尔[1] 托合提阿吉·巴拉提 阿不都外力·吾守尔[1] Li Chunxing;Dilidan·Naisier;Tuohetiaji·Balati;Abuduwaili·Wushouer(Department of Gastrointestinal Surgery,the First Affiliated Hospital of Xinjiang Medical University,Urumqi Xinjiang 830000,China)

机构地区：[1]新疆医科大学第一附属医院胃肠外科,新疆乌鲁木齐830000

出　　处：《遵义医科大学学报》2024年第9期857-867,共11页Journal of Zunyi Medical University

基　　金：新疆维吾尔自治区自然科学基金资助项目(NO:2023D01C99)。

摘　　要：目的探究v-myb禽成髓细胞病病毒癌基因同源物(MYB)转录调控线粒体裂变调节因子2(MTFR2)对胃癌(GC)细胞顺铂(DDP)耐药性的影响及分子作用机制。方法TCGA数据库分析GC中差异mRNA并预测上游调控分子,qRT-PCR检测MTFR2和MYB的表达,双荧光素酶和染色质免疫共沉淀(ChIP)实验验证MTFR2和MYB的调控关系,细胞计数盒8(CCK-8)检测细胞活力并计算IC_(50)值,流式细胞术检测细胞周期和细胞凋亡,彗星实验检测DNA损伤,蛋白质免疫印迹法检测DNA损伤相关蛋白(γ-H2AX、ATM、p-ATM)的表达。结果MTFR2在GC组织和细胞中显著高表达,敲低MTFR2能够降低细胞增殖,阻滞S期,诱导细胞凋亡,促进DNA损伤和DDP敏感性。生信预测MTFR2存在上游转录因子MYB,MYB在GC组织和细胞中的表达显著上调,双荧光素酶和ChIP验证了MTFR2启动子区域与MYB的结合关系。回复实验发现进一步过表达MTFR2能够逆转敲低MYB对GC细胞增殖和DDP耐药性的抑制作用。结论MYB上调MTFR2的表达通过DNA损伤途径促进GC细胞增殖和DDP耐药,表明靶向MYB/MTFR2调控轴可能是克服GC DDP耐药性的潜在途径。Objective To investigate the effect and molecular mechanism of v-myb avian myeloblastosis viral oncogene homolog(MYB)transcriptional regulation of mitochondrial fission regulator 2(MTFR2)on cisplatin(DDP)resistance in gastric cancer(GC)cells.Methods TCGA database was used to analyze the differential mRNAs in GC and predict its upstream regulatory molecules,qRT-PCR was used to detect the expression of MTFR2 and MYB.Dual-luciferase and Chromatin Immunoprecipitation(ChIP)experiments was used to verify the regulatory relationship between MTFR2 and MYB.Cell Counting Kit-8(CCK-8)was used to detect the cell viability and calculate IC 50 value.Flow cytometry was used to detect cell cycle and apoptosis.Comet assay was used to detect DNA damage,and protein immunoblotting was used to detect the expression of DNA damage-associated proteins(γ-H2AX,ATM,p-ATM).Results MTFR2 was significantly overexpressed in GC tissues and cells,and knockdown of MTFR2 reduced cell proliferation,blocked S phase,induced apoptosis,and promoted DNA damage and DDP sensitivity.Bioinformatics predicted the presence of the upstream transcription factor MYB in MTFR2,the expression of MYB was significantly upregulated in GC tissues and cells,and dual luciferase and ChIP verified the binding relationship between the MTFR2 promoter region and MYB.Response experiments revealed that further overexpression of MTFR2 was able to reverse the inhibitory effect of knockdown of MYB on GC cell proliferation and DDP resistance.Conclusion Upregulation of MTFR2 expression by MYB promotes GC cell proliferation and DDP resistance through the DNA damage pathway,suggesting that targeting the MYB/MTFR2 regulatory axis may be a potential pathway to overcome DDP resistance in GC.

关 键 词：V-myb禽成髓细胞瘤病毒癌基因同源物 线粒体裂变调节因子2 DNA损伤修复 胃癌 顺铂耐药 

分 类 号：R735.2[医药卫生—肿瘤]