本文選題：甲狀腺癌　切入點：miRNA　出處：《吉林大學》2016年博士論文

【摘要】：甲狀腺癌是世界范圍內(nèi)最常見的內(nèi)分泌系統(tǒng)惡性腫瘤。甲狀腺癌最常見的病理類型是甲狀腺乳頭狀癌(PTC),約占甲狀腺癌總數(shù)的80%。盡管大多數(shù)甲狀腺癌具有良好的預后,5年生存率在95%以上,但隨著甲狀腺癌發(fā)病率的逐年增高,促使甲狀腺癌發(fā)生和發(fā)展的分子生物學事件尚不明確,遺傳學病因有待進一步研究。目前術前診斷甲狀腺癌的金標準是超聲引導下細針穿刺細胞學檢查(FNAB),但尚有20-30%的FNA結果顯示為不確定的/可疑的,這部分患者需要進行診斷性手術以明確惡性或良性。分子標志物的應用將有望協(xié)助FNAB提高術前對甲狀腺癌的診斷能力。micro RNA(mi RNA)是由18-22個核苷酸組成的單鏈RNA,通過與靶基因的非轉錄區(qū)結合,調(diào)控靶基因的表達,進而影響細胞生化過程和信號傳導通路,引起腫瘤的發(fā)生和發(fā)展。mi RNA在腫瘤發(fā)生發(fā)展過程中具有關鍵作用的已達成普遍共識,然而甲狀腺腫瘤中mi RNA的表達信息及其與靶基因之間的相互作用尚不明確。據(jù)此,本研究納入了TCGA數(shù)據(jù)庫中499例甲狀腺乳頭狀癌樣本和58例甲狀腺癌旁對照組織樣本,分析了甲狀腺癌相關mi RNA和基因的表達譜,得到了38個甲狀腺癌相關差異表達的mi RNA和2731個差異表達的基因。為了尋找參與甲狀腺癌發(fā)生發(fā)展的mi RNA和基因,我們對2731個差異表達的基因進行KEGG Pathway聚類分析,得到了82個腫瘤信號通路相關的基因。通過靶基因預測,其中16個mi RNA和20個基因具有靶向調(diào)控關系,形成參與甲狀腺癌發(fā)生發(fā)展的調(diào)控網(wǎng)絡。為了探求具有診斷能力的分子標志物,我們通過繪制ROC曲線對甲狀腺癌相關且差異表達的mi RNA和基因進行評估。在差異表達的mi RNA中,上調(diào)的mi R-34a、mi R-221、mi R-222、mi R-146b,和下調(diào)的mi R-9-1、mi R-486、mi R-451、mi R-144、mi R-9-2、mi R-195,AUC值均大于0.9,并且敏感性或特異性均大于80%,適宜作為診斷標志物;其中mi R-34a的敏感性和特異性均大于90%,是具有突出診斷潛能的分子標志物。在差異表達的基因中,上調(diào)的CDKN2B、CCND1、TGFA、ELFN1、BID、DVL1、BCL2L1、RARA、BAX、E2F1、PDGFA、RXRG、MET、LPAR5、ITGA2,和下調(diào)的BCL2、TRAF6、GLI1、KIT、LPAR1、WNT11、RARB、PRKX、FGFR2,AUC值高于0.9,并且敏感性和特異性均高于80%,適宜作為診斷標志物;其中,上調(diào)的CDKN2B、CCND1和TGFA,敏感性和特異性均高于90%,是理想的獨立分子診斷標志物。為了進一步提高分子標志物對甲狀腺癌的診斷能力,我們將單個的分子標志物作以組合,組合的形式為mi RNA靶基因的聯(lián)合,其依據(jù)為mi RNA與靶基因通常是負向調(diào)控的關系,即若某mi RNA表達上調(diào),其靶基因相應地表達下調(diào);若以二者表達水平值作比,可能將mi RNA上調(diào)水平和靶基因下調(diào)水平放大,以增加鑒別效能。在篩選出的具有良好診斷能力的mi RNA和基因之間,有20對具有靶向調(diào)控關系,其中有18/20的組合AUC值比單獨mi RNA或基因有所提高。特別是mi R-34a/BCL2組合,AUC值由單獨的0.945和0.955提高到0.981,敏感性由單獨的92.2%和88.4%提高到95%,特異性由單獨的94.8%和93.1%提高到96.6%,可以認為是一個理想的腫瘤組合標志物。為了探索差異表達的mi RNA和基因是否具有侵襲特征的臨床相關性,我們將差異表達的mi RNA和基因逐一進行臨床相關性分析,包括腺體外浸潤、淋巴結轉移和TNM分期等。結果顯示,在38個差異表達的mi RNA,mi R-146b、mi R-222、mi R-21、mi R-375、mi R-652與淋巴結轉移、腺體外浸潤和高TNM分期都相關。mi R-221、mi R-181a-1、mi R-152與淋巴結轉移相關;mi R-221、mi R-34a、mi R-424、mi R-138-1、mi R-138-2、mi R-20b、mi R-152、mi R-486、hsa-mir-451、mi R-153、mi R-7-2腺體外浸潤相關;mi R-34a、mi R-424、mi R-138-1、mi R-20b、mi R-486、hsa-mir-451、mi R-7-2與高TNM分期相關。在82個差異表達的基因中,MET、TGFBR1、RARA、BAX、LAMC2、BCL2、PDGFRA、BCL2L1、TCF7L1、CDKN2B、KIT、DAPK2、LPAR5、FGF11、PAX8、FGFR2、PRKACB、GNAI1、RUNX1、GNG7、TGFB1、HSP90B1、TRAF6、ITGA3、LAMB3與淋巴結轉移、腺體外浸潤和高TNM分期都相關。PTCH2、EGF、DVL1、ITGA2B、PRKCA、RXRG、WNT3、AXIN2、TGFA與淋巴結轉移相關;DVL1、ITGA2、BID、FZD5、ITGA2B、PTCH2、T GFA、AXIN2、CDKN1A、RARB、WNT11、GLI3、RUNX1T1與高TNM分期相關。綜上,本研究基于TCGA數(shù)據(jù)庫的大樣本生物信息數(shù)據(jù),分析了甲狀腺癌相關mi RNA和基因表達譜,并形成了甲狀腺癌中mi RNA與靶基因的調(diào)控網(wǎng)絡。部分差異表達的mi RNA和基因顯示出良好的診斷潛能和臨床相關性,可在臨床實踐中指導甲狀腺癌的診斷、風險分層、制定治療策略和評估預后等。
[Abstract]:Thyroid cancer is the most common endocrine system in the world. The most common pathological type of thyroid cancer is papillary thyroid carcinoma (PTC), accounting for the total number of thyroid cancer 80%. although most thyroid carcinoma has a good prognosis, 5 years survival rate is more than 95%, but with the increasing incidence of thyroid cancer. The thyroid cancer occurrence and development of molecular biological events is unclear, genetic etiology needs further study. The current gold standard for preoperative diagnosis of thyroid cancer is the ultrasound guided fine needle aspiration cytology (FNAB), but there is still 20-30% FNA results is uncertain / suspected, these patients require diagnostic the operation to identify benign or malignant. The application of molecular markers is expected to help FNAB RNA.Micro diagnosis for thyroid cancer to improve the preoperative (MI RNA) is composed of 18-22 nucleotides. The single stranded RNA, through the combination of untranslated region and target genes, regulating the expression of target genes, thereby affecting cellular biochemical processes and signal transduction pathways, which plays a key role in tumor occurrence and development of.Mi RNA in tumor development process has reached a general consensus, but the interaction between the expression of MI in thyroid tumor information RNA and its target gene is not clear. Therefore, the research into the TCGA database in 499 cases of thyroid papillary carcinoma samples and 58 cases of thyroid cancer adjacent normal tissue samples, analysis of RNA and MI for radical related thyroid cancer expression, obtained the expression of MI RNA expression in 38 thyroid cancer related differences and 2731 genes involved in MI and RNA. In order to find the genetic development of thyroid carcinoma, we analysis KEGG Pathway cluster on the expression of 2731 genes differentially, obtained 82 tumor signal Pathway related genes. The target gene prediction, with 16 mi RNA and 20 genes, which targeted regulation, regulatory networks involved in the formation of the occurrence and development of thyroid cancer. In order to explore the molecular markers with diagnostic capabilities, we assessed by MI and RNA gene expression and the difference of thyroid cancer ROC curve in the MI RNA. The differential expression, up regulation of MI R-34a, MI R-221, MI R-222, MI R-146b, MI R-9-1 MI and down-regulation of R-486, MI, R-451, MI, R-144, MI, R-9-2, MI, R-195, AUC values are greater than 0.9, and the specificity and sensitivity were higher than 80%, suitable as a diagnostic marker MI; the sensitivity and specificity of R-34a were greater than 90%, is a molecular marker with prominent diagnostic potential. The differentially expressed genes, upregulation of CDKN2B, CCND1, TGFA, ELFN1, BID, DVL1, BCL2L1, RARA, BAX, E2F1, PDGFA, RXRG, MET, LPAR5, ITGA2, and down the BCL2, TRAF6, GLI1, KIT, LPAR1, WNT11, RARB, PRKX, FGFR2, AUC higher than 0.9, and the sensitivity and specificity were higher than 80%, suitable as a diagnostic marker; wherein, upregulation of CDKN2B, CCND1 and TGFA, the sensitivity and specificity were higher than 90%, is the ideal independent molecular markers. In order to further improve the molecular markers of thyroid cancer diagnosis ability, we will be single molecular markers in combination, combination of the form of joint mi RNA target gene, which is based on MI RNA and target gene is usually negative relation to regulation of expression, if a mi RNA, the corresponding target genes the expression levels of expression in the two cut; value ratio, MI may upregulate RNA levels and lower levels of target gene amplification, in order to increase the identification efficiency. Among the screened with good diagnostic ability of MI and RNA gene, 20 of the targeted regulation relationship, including 18/20 The combined AUC value increased more than Mi alone or RNA gene. Especially mi R-34a/BCL2, the AUC value from 0.945 separate and 0.955 increased to 0.981, the sensitivity increased from 92.2% and 88.4% to 95% alone, the specificity increased by 94.8% and 93.1% to 96.6% separate, can be considered an ideal combination of tumor markers in order to explore the differences of expression. Whether the MI gene has RNA and clinical relevance of invasive features, we will mi RNA and differential gene expression by clinical correlation analysis, including glandular infiltration, lymph node metastasis and TNM stage. The results showed that MI RNA, the 38 differentially expressed mi R-146b, MI R-222. Mi R-21, MI R-375, MI R-652 and lymph node metastasis, infiltration glands and high TNM stage are.Mi R-221, MI R-181a-1, MI R-152 was correlated with lymph node metastasis; MI R-221, MI R-34a, MI R-424, MI R-138-1, MI R-138-2, MI R-20b, MI R-152 ,mi R-486,hsa-mir-451,mi R-153,mi R-7-2鑵轟綋澶栨蹈娑︾浉鍏，

本文編號：1708586