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

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ErbB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--p、miR--p可能分别通过调控靶基因PRKCB、MAPK、PAK、PRKAA、CCNE、IGFR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 ErbB 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,IGF1R.