表达谱芯片筛查非小细胞肺癌细胞系A549放疗抵抗差异基因
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摘要
目的建立非小细胞肺癌(NSCLC)A549放疗抵抗模型,表达谱芯片技术筛查放疗抵抗差异基因。方法 A549细胞系长期间歇照射诱导建立的放疗抵抗细胞模型;表达谱基因芯片筛查放疗抵抗细胞株与敏感株全基因组RNA表达差异;分析2倍以上差异的基因(P<0.05),分别进行生物学信息基因分类(GO)和信息通路(Pathway)分析。结果表达谱芯片显示,抵抗株与敏感株相比,差异表达基因为1410个(733个上调,677个下调);GO分析显示,主要与细胞周期、DNA修复有关;通路显著性富集分析显示,丝裂原活化蛋白激酶(MAPK)和磷脂酰肌醇-3-激酶(PI3K)等多条信号通路激酶出现显著性差异。结论多种基因和信号通路参与了放疗抵抗过程。
Objective To investigate the radioresistance factors in non-small cell lung cancer(NSCLC)cell line A549, and provide new targets for radiotherapy sensitization drugs development. Methods Establish the stable model of radioresistant NSCLC cell line A549 under irradiation; investigate the whole-transcriptome alteration of radioresistance cell line and radiosensitive cell line using gene expression microarray; perform bioinformatic approaches gene ontology(GO) analysis and Pathway analysis. Results The expression profile microarray showed that 1410 differentially expressed genes(733 up-regulated and 677 down-regulated) were detected in resistant and sensitive strains; GO analysis showed that it was mainly related to cell cycle and DNA replication; Pathway significant enrichment analysis showed that mitogen-activated protein kiase(MAPK) signaling pathway, phosphatidylinositol 3-kinase/protein kinase B(PI3 K/Akt) signaling pathway, were mainly associated with radioresistance. Conclusion Multiple genes and signaling pathways are involved in radioresistance, further studies are needed to investigate the radioresistance factors, which could provide new targets for radiotherapy sensitization drugs development.
Objective To investigate the radioresistance factors in non-small cell lung cancer(NSCLC)cell line A549, and provide new targets for radiotherapy sensitization drugs development. Methods Establish the stable model of radioresistant NSCLC cell line A549 under irradiation; investigate the whole-transcriptome alteration of radioresistance cell line and radiosensitive cell line using gene expression microarray; perform bioinformatic approaches gene ontology(GO) analysis and Pathway analysis. Results The expression profile microarray showed that 1410 differentially expressed genes(733 up-regulated and 677 down-regulated) were detected in resistant and sensitive strains; GO analysis showed that it was mainly related to cell cycle and DNA replication; Pathway significant enrichment analysis showed that mitogen-activated protein kiase(MAPK) signaling pathway, phosphatidylinositol 3-kinase/protein kinase B(PI3 K/Akt) signaling pathway, were mainly associated with radioresistance. Conclusion Multiple genes and signaling pathways are involved in radioresistance, further studies are needed to investigate the radioresistance factors, which could provide new targets for radiotherapy sensitization drugs development.
引文
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[19] Coopes A,Henry CE,Llamosas E,et al.An update of Wnt signalling in endometrial cancer and its potential as a therapeutic target[J].Endocr Relat Cancer,2018,doi:10.1530/ERC-18-0112.
[20] Wang Q,Feng F,Wang J,et al.Liver X receptor activation reduces gastric cancer cell proliferation by suppressing Wnt signalling via LXRβ relocalization[J].J Cell Mol Med,2019,23(2):789-797.
[21] Luo X,Ye S,Jiang Q,et al.Wnt inhibitory factor-1-mediated autophagy inhibits Wnt/β-catenin signaling by downregulating dishevelled-2 expression in non-small cell lung cancer cells[J].Int J Oncol,2018,53(2):904-914.
[22] Yang F,Li Y,Liu B,et al.Cancer stem cell-like population is preferentially suppressed by EGFR-TKIs in EGFR-mutated PC-9 tumor models[J].Exp Cell Res,2018,362(1):195-202.
[2] Yan B,Liu S,Shi Y,et al.Activation of AhR with nuclear IKKα regulates cancer stem-like properties in the occurrence of radioresistance[J].Cell Death Dis,2018,9(5):490.
[3] Zhao WY,Sun XCh.Progression of tumor radiosensitization[J].Chinese Clinical Oncology,2012,17(7):655-659.(in Chinese)赵维勇,孙新臣.放射治疗增敏的研究现状[J].临床肿瘤学杂志,2012,17(7):655-659.
[4] Fu Z,Chen D,Cheng H,et al.Hypoxia-inducible factor-1α protects cervical carcinoma cells from apoptosis induced by radiation via modulation of vascular endothelial growth factor and p53 under hypoxia[J].Med Sci Monit,2015,21:318-325.
[5] Ausborn NL,Le QT,Bradley JD,et al.Molecular profiling to optimize treatment in non-small cell lung cancer:a review of potential molecular targets for radiation therapy by the translational research program of the radiation therapy oncology group[J].Int J Radiat Oncol Biol Phys,2012,83(4):e453-464.
[6] Graves EE,Maity A,Le QT.The tumor microenvironment in non-small-cell lung cancer[J].Semin Radiat Oncol,2010,20(3):156-163.
[7] Gu XZh,Yin WB,Yu ZH,et al.Radiation Oncology[M].Beijing:Peking Union Medical College Press,2008:261-263.(in Chinese)谷铣之,殷蔚伯,余子豪,等.肿瘤放射治疗学[M].北京:协和医科大学出版社,2018:261-263.
[8] Yang HJ,Kim N,Seong KM,et al.Investigation of radiation-induced transcriptome profile of radioresistant non-small cell lung cancer A549 cells using RNA-seq[J].PLoS One,2013,8(3):e59319.
[9] Sun J,Liu NB,Qu ChH,et al.Identifying the genetic pattern of conventional fractionated and hypofractionated radiotherapy using whole genome expression microarray in a non-small-cell lung cancer cell line[J].Chinese Journal of Clinical Oncology,2013,40(21):1280-1283.(in Chinese)孙健,刘宁波,曲晨慧,等.全基因组表达谱芯片筛选非小细胞肺癌常规分割和大分割放疗差异基因的初步研究[J].中国肿瘤临床,2013,40(21):1280-1283.
[10] Borchiellini D,Etienne-Grimaldi MC,Thariat J,et al.The impact of pharmacogenetics on radiation therapy outcome in cancer patients.A focus on DNA damage response genes[J].Cancer Treat Rev,2012,38(6):737-759.
[11] Yang L,Yang G,Ding Y,et al.Inhibition of PI3K/AKT signaling pathway radiosensitizes pancreatic cancer cells with ARID1A deficiency in vitro[J].J Cancer,2018,9(5):890-900.
[12] Xu S,Li Y,Lu Y,et al.LZTS2 inhibits PI3K/AKT activation and radioresistance in nasopharyngeal carcinoma by interacting with p85[J].Cancer Lett,2018,420:38-48.
[13] Saavedra D,Neninger E,Rodriguez C,et al.CIMAvax-EGF:Toward long-term survival of advanced NSCLC[J].Semin Oncol,2018,45(1-2):34-40.
[14] Concha-Benavente F,Ferris RL.Reversing EGFR mediated immunoescape by targeted monoclonal antibody therapy[J].Front Pharmacol,2017,8:332.
[15] Shi Y,Liu N,Lai W,et al.Nuclear EGFR-PKM2 axis induces cancer stem cell-like characteristics in irradiation-resistant cells[J].Cancer Lett,2018,422:81-93.
[16] Majidinia M,Aghazadeh J,Jahanban-Esfahlani R,et al.The roles of Wnt/β-catenin pathway in tissue development and regenerative medicine[J].J Cell Physiol,2018,233(8):5598-5612.
[17] Nusse R,Clevers H.Wnt/β-catenin signaling,disease,and emerging therapeutic modalities[J].Cell,2017,169(6):985-999.
[18] Ahmad Ⅰ,Sansom OJ.Role of Wnt signalling in advanced prostate cancer[J].J Pathol,2018,245(1):3-5.
[19] Coopes A,Henry CE,Llamosas E,et al.An update of Wnt signalling in endometrial cancer and its potential as a therapeutic target[J].Endocr Relat Cancer,2018,doi:10.1530/ERC-18-0112.
[20] Wang Q,Feng F,Wang J,et al.Liver X receptor activation reduces gastric cancer cell proliferation by suppressing Wnt signalling via LXRβ relocalization[J].J Cell Mol Med,2019,23(2):789-797.
[21] Luo X,Ye S,Jiang Q,et al.Wnt inhibitory factor-1-mediated autophagy inhibits Wnt/β-catenin signaling by downregulating dishevelled-2 expression in non-small cell lung cancer cells[J].Int J Oncol,2018,53(2):904-914.
[22] Yang F,Li Y,Liu B,et al.Cancer stem cell-like population is preferentially suppressed by EGFR-TKIs in EGFR-mutated PC-9 tumor models[J].Exp Cell Res,2018,362(1):195-202.