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

【摘要】：甲狀腺癌是世界范圍內(nèi)最常見(jiàn)的內(nèi)分泌系統(tǒng)惡性腫瘤。甲狀腺癌最常見(jiàn)的病理類(lèi)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(yù)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(xué)事件尚不明確,遺傳學(xué)病因有待進(jìn)一步研究。目前術(shù)前診斷甲狀腺癌的金標(biāo)準(zhǔn)是超聲引導(dǎo)下細(xì)針穿刺細(xì)胞學(xué)檢查(FNAB),但尚有20-30%的FNA結(jié)果顯示為不確定的/可疑的,這部分患者需要進(jìn)行診斷性手術(shù)以明確惡性或良性。分子標(biāo)志物的應(yīng)用將有望協(xié)助FNAB提高術(shù)前對(duì)甲狀腺癌的診斷能力。micro RNA(mi RNA)是由18-22個(gè)核苷酸組成的單鏈RNA,通過(guò)與靶基因的非轉(zhuǎn)錄區(qū)結(jié)合,調(diào)控靶基因的表達(dá),進(jìn)而影響細(xì)胞生化過(guò)程和信號(hào)傳導(dǎo)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(guò)程中具有關(guān)鍵作用的已達(dá)成普遍共識(shí),然而甲狀腺腫瘤中mi RNA的表達(dá)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(kù)中499例甲狀腺乳頭狀癌樣本和58例甲狀腺癌旁對(duì)照組織樣本,分析了甲狀腺癌相關(guān)mi RNA和基因的表達(dá)譜,得到了38個(gè)甲狀腺癌相關(guān)差異表達(dá)的mi RNA和2731個(gè)差異表達(dá)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(duì)2731個(gè)差異表達(dá)的基因進(jìn)行KEGG Pathway聚類(lèi)分析,得到了82個(gè)腫瘤信號(hào)通路相關(guān)的基因。通過(guò)靶基因預(yù)測(cè),其中16個(gè)mi RNA和20個(gè)基因具有靶向調(diào)控關(guān)系,形成參與甲狀腺癌發(fā)生發(fā)展的調(diào)控網(wǎng)絡(luò)。為了探求具有診斷能力的分子標(biāo)志物,我們通過(guò)繪制ROC曲線對(duì)甲狀腺癌相關(guān)且差異表達(dá)的mi RNA和基因進(jìn)行評(píng)估。在差異表達(dá)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(biāo)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(biāo)志物。在差異表達(dá)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(biāo)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(dú)立分子診斷標(biāo)志物。為了進(jìn)一步提高分子標(biāo)志物對(duì)甲狀腺癌的診斷能力,我們將單個(gè)的分子標(biāo)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(fù)向調(diào)控的關(guān)系,即若某mi RNA表達(dá)上調(diào),其靶基因相應(yīng)地表達(dá)下調(diào);若以二者表達(dá)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(duì)具有靶向調(diào)控關(guān)系,其中有18/20的組合AUC值比單獨(dú)mi RNA或基因有所提高。特別是mi R-34a/BCL2組合,AUC值由單獨(dú)的0.945和0.955提高到0.981,敏感性由單獨(dú)的92.2%和88.4%提高到95%,特異性由單獨(dú)的94.8%和93.1%提高到96.6%,可以認(rèn)為是一個(gè)理想的腫瘤組合標(biāo)志物。為了探索差異表達(dá)的mi RNA和基因是否具有侵襲特征的臨床相關(guān)性,我們將差異表達(dá)的mi RNA和基因逐一進(jìn)行臨床相關(guān)性分析,包括腺體外浸潤(rùn)、淋巴結(jié)轉(zhuǎn)移和TNM分期等。結(jié)果顯示,在38個(gè)差異表達(dá)的mi RNA,mi R-146b、mi R-222、mi R-21、mi R-375、mi R-652與淋巴結(jié)轉(zhuǎn)移、腺體外浸潤(rùn)和高TNM分期都相關(guān)。mi R-221、mi R-181a-1、mi R-152與淋巴結(jié)轉(zhuǎn)移相關(guān);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(rùn)相關(guān);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(guān)。在82個(gè)差異表達(dá)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(jié)轉(zhuǎn)移、腺體外浸潤(rùn)和高TNM分期都相關(guān)。PTCH2、EGF、DVL1、ITGA2B、PRKCA、RXRG、WNT3、AXIN2、TGFA與淋巴結(jié)轉(zhuǎn)移相關(guān);DVL1、ITGA2、BID、FZD5、ITGA2B、PTCH2、T GFA、AXIN2、CDKN1A、RARB、WNT11、GLI3、RUNX1T1與高TNM分期相關(guān)。綜上,本研究基于TCGA數(shù)據(jù)庫(kù)的大樣本生物信息數(shù)據(jù),分析了甲狀腺癌相關(guān)mi RNA和基因表達(dá)譜,并形成了甲狀腺癌中mi RNA與靶基因的調(diào)控網(wǎng)絡(luò)。部分差異表達(dá)的mi RNA和基因顯示出良好的診斷潛能和臨床相關(guān)性,可在臨床實(shí)踐中指導(dǎo)甲狀腺癌的診斷、風(fēng)險(xiǎn)分層、制定治療策略和評(píng)估預(yù)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(hào)：1708586