卵巢浆液性囊腺癌化疗耐药和化疗敏感组织中的miRNAs差异表达谱检测及生物信息学分析
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摘要
目的:寻找卵巢浆液性囊腺癌化疗耐药及化疗敏感组织中microRNA的差异表达,为研究上皮性卵巢癌化疗耐药产生的分子机制及逆转其化疗耐药提供理论基础和依据。方法:首先,应用microRNA表达谱基因芯片寻找卵巢浆液性囊腺癌化疗耐药及化疗敏感组织中差异表达的microRNAs;其次,选取部分差异表达的microRNAs应用RT-PCR实验检测技术在扩大的组织样本中进行验证;最后,运用生物信息学技术对符合要求的microRNAs进行相关生物信息学分析。结果:在microRNA表达谱基因芯片中筛选出与卵巢浆液性囊腺癌化疗耐药相关的miR-27a-3p、miR-141-3p、miR-200b-3p等17个表达上调的miRNAs和miR-93-3p、miR-497-5p、miR-675-3p等15个表达下调的miRNAs。上调miRNAs预测的靶基因GO(BP模块)分析结果显示凋亡信号通路、Notch信号通路和凋亡过程,KEGG Pathway分析结果显示Wnt信号通路、PI3K-Akt信号通路和Erb B信号通路可能与化疗耐药存在相关性。下调miRNAs预测的靶基因GO(BP模块)分析结果显示负调控凋亡过程、Wnt信号通路生物学过程,KEGG Pathway分析结果显示PI3K-Akt信号通路、细胞循环可能与化疗耐药存在相关性。结论:MicroRNA表达谱基因芯片及生物信息学技术能为研究上皮性卵巢癌化疗耐药产生机制提供新的思路。miR-27a-3p、miR-141-3p、miR-200b-3p、miR-93-3p、miR-497-5p、miR-675-3p可能分别通过调控靶基因PRKCB、MAP2K4、PAK2、PRKAA1、CCNE1、IGF1R参与卵巢浆液性囊腺癌化疗耐药。
Objective: To research the differential microRNA exprsession in ovarian serous carcinoma chemotherapy resistance and sensitive tissues in order to provide theory basis for studying molecular mechanism of chemotherapy resistance and reversing it. Methods: Firstly,using the microarray to search the differential microRNAs exprsession in ovarian serous carcinoma chemotherapy resistance and sensitive tissue. Secondly,validating some candidate miRNAs in expanding samples with Real-time PCR technology. Lastly,analyzing the significant microRNA with bioinformatic software. Results: MicroRNAs were related to ovarian serous carcinoma chemotherapy resistance by the microarray.There were 17 miRNAs( such as miR-27a-3p,miR-141-3p and miR-200b-3p) expression up-regulated,and 15 miRNAs( such as miR-93-3p,miR-497-5p and miR-675-3p) expression down-regulated. The result of up-regulated miRNAs targeting genes by GO analyzing( biological process) included instrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator,Notch signaling pathway,apoptotic process and the result of it by KEGG Pathway analyzing included Wnt signaling pathway,PI3K/Akt signaling pathway and Erb B signaling pathway. The result of down-regulated miRNAs targeting genes by GO analyzing( biological process) included negative regulation of apoptotic process,Wnt signaling pathway and the result of it by KEGG Pathway analyzing included PI3K/Akt signaling pathway and cell cycle. All may have correlation with chemoresistance. Conclusion: The microarray and bioinformatic technology could provide a newapproach for researching epithelial ovarian cancer chemotherapy resistance. miR-27a-3p,miR-141-3p,miR-200b-3p,miR-93-3p,miR-497-5p,miR-675-3p may play an important role in epithelial ovarian cancer chemotherapy resistance via their targeting gene PRKCB,MAP2K4,PAK2,PRKAA1,CCNE1,IGF1 R.
Objective: To research the differential microRNA exprsession in ovarian serous carcinoma chemotherapy resistance and sensitive tissues in order to provide theory basis for studying molecular mechanism of chemotherapy resistance and reversing it. Methods: Firstly,using the microarray to search the differential microRNAs exprsession in ovarian serous carcinoma chemotherapy resistance and sensitive tissue. Secondly,validating some candidate miRNAs in expanding samples with Real-time PCR technology. Lastly,analyzing the significant microRNA with bioinformatic software. Results: MicroRNAs were related to ovarian serous carcinoma chemotherapy resistance by the microarray.There were 17 miRNAs( such as miR-27a-3p,miR-141-3p and miR-200b-3p) expression up-regulated,and 15 miRNAs( such as miR-93-3p,miR-497-5p and miR-675-3p) expression down-regulated. The result of up-regulated miRNAs targeting genes by GO analyzing( biological process) included instrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator,Notch signaling pathway,apoptotic process and the result of it by KEGG Pathway analyzing included Wnt signaling pathway,PI3K/Akt signaling pathway and Erb B signaling pathway. The result of down-regulated miRNAs targeting genes by GO analyzing( biological process) included negative regulation of apoptotic process,Wnt signaling pathway and the result of it by KEGG Pathway analyzing included PI3K/Akt signaling pathway and cell cycle. All may have correlation with chemoresistance. Conclusion: The microarray and bioinformatic technology could provide a newapproach for researching epithelial ovarian cancer chemotherapy resistance. miR-27a-3p,miR-141-3p,miR-200b-3p,miR-93-3p,miR-497-5p,miR-675-3p may play an important role in epithelial ovarian cancer chemotherapy resistance via their targeting gene PRKCB,MAP2K4,PAK2,PRKAA1,CCNE1,IGF1 R.
引文
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[15]Suzuki HI,Yamagata K,Sugimoto K,et al.Modulation of microRNA processing by p53[J].Nature,2009,460(7254):529-533.
[16]Liu N,Zeng J,Zhang XM,et al.Involvement of miR-200a in chemosensitivity regulation of ovarian cancer[J].Natl Med J China,2014,94(2):2148-2151.[刘娜,曾洁,张小媚,等.miR-200a参与调控卵巢癌化疗敏感性的作用及其机制[J].中华医学杂志,2014,94(2):2148-2151.]
[17]Wang Sinian,Lv Jin,Zhang Qiaoyun,et al.miRNA and invasion and metastasis of cancer[J].Modern Oncology,2016,24(3):485-487.[王思念,吕进,张巧云,等.miRNA与肿瘤侵袭转移[J].现代肿瘤医学,2016,24(3):485-487.]
[18]Zou D,Wang D,Li R,et al.MiR-197 induces Taxol resistance in human ovarian cancer cells by regulating NLK[J].Tumour Biol,2015,36(9):6725-6732.
[19]Chen S,Jiao JW,Zhao Y,et al.MicroRNA-133b targets glutathione S-transferaseπexpression to increase ovarian cancer cell sensitivity to chemotherapy drugs[J].Drug Des Devel Ther,2015,9:5225-5235.
[20]Tao T,Wang M.Screening of differentially ecpression miRNA in chemoresistant and sensitive ovarian cancer cells and the identification of the tissues[J].J China Medical University,2013,42(2):118-122.[陶陶,王敏.化疗耐药及敏感卵巢癌细胞差异表达microRNA的筛查与组织鉴定[J].中国医科大学学报,2013,42(2):118-122.]
[2]Siegel R,Naishadham D,Jemal A.Cancer statistics,2012[J].CA Cancer J Clin,2012,62(1):10-29.
[3]Fotopoulou C,Savvatis K.Primary radical surgery in elderly patients with epithelial ovarian cancer analysis of surgical outcome and long-team survival[J].Int J Gynecol Cancer,2010,20(1):34-40.
[4]Shen K,Cui H,Feng YJ.Common gynecologic malignant tumor diagnosis and treatment guidelines[M].Beijing:People's Health Publishing House,2014:103.[沈铿,崔恒,丰有吉.常见妇科恶性肿瘤诊治指南[M].北京:人民卫生出版社,2014:103.]
[5]Groeneweg JW,Foster R,Growdon WB,et al.Notch signaling in serous ovarian cancer[J].J Ovarian Res,2014,7:95.
[6]Gupta N,Xu Z,EI-Sehemy A,et al.Notch3 induces epithelialmesenchymal transition and attenuates carboplatin-induced apoptosis in ovarian cancer cells[J].Gynecol Oncol,2013,130(1):200-206.
[7]Bagic-Onder T,Wakimoto H,Anderegg M,et al.A dual PI3K/m TOR inhibitor,PI-103,cooperates with stem cell-delivered TRAIL in experimental glioma models[J].Cancer Res,2011,71(1):154-163.
[8]Shi XY,Cai XJ,Lei JX,et al.Reversal effect of PI3K/Akt pathway inhibitor LY294002 on multidrug resistance of ovarian cancer cell line A2780/Taxol[J].Chin J Cancer,2007,27(4):343-347.[石小燕,蔡晓军,类建翔,等.PI3K/Akt抑制剂LY294002对卵巢癌细胞A2780/Taxol多药耐药性的逆转作用[J].癌症,2007,27(4):343-347.]
[9]Arend RC,Straughn JM JR.The Wnt/β-catenin pathway in ovarian cancer:A review[J].Gynecol Oncol,2013,131(3):772-779.
[10]Zhang J,Chen AP,Qi YY,et al.Effect of RNA-mediated EGFR gene on resistance to cisplatin of multidrug-resistance ovarian cancer[J].Chin J Cancer Prevent Treatment,2010,17(5):345-348.[张娇,陈爱平,戚玉言,等.RNA干扰靶向沉默EGFR基因对卵巢癌耐药细胞凋亡影响的实验观察[J].中华肿瘤防治杂志,2010,17(5):345-348.]
[11]Liu L,Li L.miRNA-mRNA different expression with multidrug resistance in epithelial ovarian cancer[D].Nanning:Guangxi Medical University,2014:40-41.[刘玲,李力.miRNA-mRNA差异表达与卵巢癌多药耐药关系的研究[D].南宁:广西医科大学,2014:40-41.]
[12]Calin GA,Sevignani C,Dumitru CD,et al.Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers[J].Proc Natl Acad Sic USA,2004,101(9):2999-3004.
[13]Lujambio A,Lowe SW.The microcosmos of cancer[J].Nature,2012,482(7385):347-355.
[14]Png KJ,Halberg N,Yoshida M,et al.A microRNA regulon that mediates endothelial recruitment and metastasis by cancer cells[J].Nature,2012,481(7380):190-194.
[15]Suzuki HI,Yamagata K,Sugimoto K,et al.Modulation of microRNA processing by p53[J].Nature,2009,460(7254):529-533.
[16]Liu N,Zeng J,Zhang XM,et al.Involvement of miR-200a in chemosensitivity regulation of ovarian cancer[J].Natl Med J China,2014,94(2):2148-2151.[刘娜,曾洁,张小媚,等.miR-200a参与调控卵巢癌化疗敏感性的作用及其机制[J].中华医学杂志,2014,94(2):2148-2151.]
[17]Wang Sinian,Lv Jin,Zhang Qiaoyun,et al.miRNA and invasion and metastasis of cancer[J].Modern Oncology,2016,24(3):485-487.[王思念,吕进,张巧云,等.miRNA与肿瘤侵袭转移[J].现代肿瘤医学,2016,24(3):485-487.]
[18]Zou D,Wang D,Li R,et al.MiR-197 induces Taxol resistance in human ovarian cancer cells by regulating NLK[J].Tumour Biol,2015,36(9):6725-6732.
[19]Chen S,Jiao JW,Zhao Y,et al.MicroRNA-133b targets glutathione S-transferaseπexpression to increase ovarian cancer cell sensitivity to chemotherapy drugs[J].Drug Des Devel Ther,2015,9:5225-5235.
[20]Tao T,Wang M.Screening of differentially ecpression miRNA in chemoresistant and sensitive ovarian cancer cells and the identification of the tissues[J].J China Medical University,2013,42(2):118-122.[陶陶,王敏.化疗耐药及敏感卵巢癌细胞差异表达microRNA的筛查与组织鉴定[J].中国医科大学学报,2013,42(2):118-122.]