乳腺癌阿霉素耐药细胞株中转运RNA及其来源片段表达谱分析  

Expression profile analysis of transfer ribonucleic acid-derived small RNAs in adriamycin-induced breast cancer MCF-7 resistant cells

作　　者：莫冬萍 严枫[1] Mo Dongping;Yan Feng(Jiangsu Key Laboratory of Molecular and Translational Cancer Research Jiangsu Cancer Hospital&the Affiliated Cancer Hospital of Nanjing Medicine University&Jiangsu Institute of Cancer Research,Nanjing 210009,China)

机构地区：[1]江苏省肿瘤医院,南京医科大学附属肿瘤医院,江苏省肿瘤防治研究所,江苏省恶性肿瘤分子生物学及转化医学重点实验室,210009

出　　处：《中华实验外科杂志》2022年第8期1497-1500,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(82002225、81871718)。

摘　　要：目的分析阿霉素诱导的MCF-7耐药细胞(MCF-7/Adriamycin, MCF-7/ADR)中转运RNA及其来源片段(tRNA-Derived small RNAs, tDRs)表达谱变化, 筛选出调控乳腺癌阿霉素耐药的tDRs。方法采用高通量测序检测MCF-7/ADR细胞的tDRs表达谱, 筛选差异表达tDRs。基因本体和京都基因与基因组百科全书对差异tDRs进行生物学通路分析。细胞凋亡实验和实时定量聚合酶链反应检测转染后细胞凋亡能力和凋亡相关基因。两两比较采用成组t检验。结果 MCF-7/ADR细胞中73条tDRs存在差异(差异倍数>2, P<0.05)。生物学分析示tDRs的主要功能及通路和肿瘤相关, 尤其是上调的tDRs(tRF-58-75-Ala-AGC-1, tRF-58-75-Ala-AGC-5, tRF-58-75-Pro-AGG-1-M7和tRF-59-76-Tyr-GTA-4)与细胞凋亡关系密切。转染tRF-58-75-Ala-AGC-1抑制剂后, MCF-7/ADR转染组细胞凋亡率高于阴性对照组[(83.34±1.95)%比(71.12±5.01)%, t=3.937, P<0.05];MCF-7/ADR转染组细胞中B细胞淋巴瘤/白血病-2(bcl-2) mRNA表达水平低于阴性对照组(0.58±0.23比1.00±0.05, t=3.025, P<0.05);半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3、7、9 mRNA表达水平高于阴性对照组(1.29±0.11、1.50±0.16、1.22±0.07比1.00±0.05, t=4.226、5.296、4.174, P<0.05)。结论 MCF-7/ADR细胞的tDRs表达存在差异。Objective To study the changes of tRNA-derived small RNAs(tDRs)expression profile in adriamycin-induced breast cancer MCF-7 resistant cells,and screen the tDRs for adriamycin resistant breast cancer.Methods High-throughput sequencing was used to detect the expression profile of tDRs in MCF-7/adriamycin(MCF-7/ADR)cells and screen the differentially expressed tDRs.Biological pathways of differential tDRs were analyzed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes.Apoptosis test and real-time quantitative polymerase chain reaction were used to detected the apoptosis ability and apoptosis-related gene of transfected cells.GraphPad Prism 8 was carried to process the data,and pairwise comparisons were performed by group t test.Results A total of 73 tDRs were differentially expressed in MCF-7/ADR(fold change>2,P<0.05).The biological analysis showed that the main function and pathway of tDRs were related to tumor,especially the up-regulated tDRs(tRF-58-75-Ala-AGC-1,tRF-58-75-Ala-AGC-5,tRF-58-75-Pro-AGG-1-M7 and tRF-59-76-Tyr-GTA-4)were closely related to apoptosis.After transfection with tRF-58-75-Ala-AGC-1 inhibitor,the apoptosis rate of MCF-7/ADR transfection group was higher than that of negative control group[(83.34±1.95)%vs.(71.12±5.01)%,t=3.937,P<0.05].The expression level of B cell lymphoma/leukemia-2(bcl-2)mRNA in MCF-7/ADR transfection group was lower than that in negative control group(0.58±0.23 vs.1.00±0.05,t=3.025,P<0.05).The cysteinyl aspartate-specific protease(Caspase)-3,7,9 mRNA expression levels were higher than those in the negative control group(1.29±0.11,1.50±0.16,1.22±0.07 vs.1.00±0.05,t=4.226,5.296,4.174,P<0.05).Conclusion The study found that the expression of tDRs inMCF-7/ADR is different.Differentially expressed genes can provide novel clues for finding new biological targets for breast cancer.

关 键 词：转运RNA 阿霉素耐药 乳腺癌 

分 类 号：R737.9[医药卫生—肿瘤]