本文選題：胃癌 + 血漿��； 參考：《南京醫(yī)科大學(xué)》2017年碩士論文

【摘要】：背景:我國(guó)一直是胃癌(gastriccancer,GC)高發(fā)國(guó)家,每年胃癌發(fā)病數(shù)和死亡數(shù)均占世界總數(shù)的50%左右,疾病負(fù)擔(dān)嚴(yán)重,是腫瘤防治的重點(diǎn)。胃癌患者多沒(méi)有特異性癥狀,因而診斷時(shí)大多數(shù)患者已處于進(jìn)展期,進(jìn)展期胃癌預(yù)后較差,術(shù)后五年生存率低于30%,而早期胃癌預(yù)后良好,術(shù)后五年生存率可以達(dá)到90%以上,因此尋找能夠鑒別早期胃癌、且具有高敏感性和特異性的生物標(biāo)志至關(guān)重要。長(zhǎng)鏈非編碼RNAs(long non-coding RNAs,lncRNAs)是指長(zhǎng)度超過(guò)200個(gè)核苷酸且沒(méi)有蛋白編碼能力的RNA轉(zhuǎn)錄本。目前大量研究已經(jīng)表明lncRNAs在腫瘤發(fā)生發(fā)展過(guò)程中發(fā)揮著重要作用。近年來(lái),lncRNAs被發(fā)現(xiàn)可以釋放到循環(huán)中且穩(wěn)定存在,且循環(huán)lncRNAs的表達(dá)水平在腫瘤患者和健康對(duì)照體內(nèi)存在顯著差異,有可能作為腫瘤潛在生物標(biāo)志而改善腫瘤的早期診斷。目前循環(huán)lncRNAs的研究尚處于起步階段,循環(huán)lncRNA的來(lái)源及其功能還未系統(tǒng)闡明。方法:本研究采用Arraystar Human lncRNA/mRNA v3.0芯片技術(shù),檢測(cè)并分析5例胃癌病例及5例性別、年齡相匹配健康對(duì)照的全基因組血漿lncRNAs;以胃癌患者血漿中高表達(dá)最顯著的前10條lncRNAs作為本研究的候選lncRNAs,運(yùn)用實(shí)時(shí)定量 PCR(quantitative real-time polymerase chain reaction,qRT-PCR)技術(shù)進(jìn)行兩階段血漿大樣本(一階段包括50例胃癌病例血漿和50例健康對(duì)照血漿,二階段包括173例胃癌病例血漿和173例對(duì)照血漿)驗(yàn)證,通過(guò)受試者工作特征曲線(receive operating charactreristic curve,ROC)分析及計(jì)算曲線下面積(area under the curve,AUC),評(píng)價(jià)單個(gè)或一組lncRNAs對(duì)胃癌診斷的應(yīng)用價(jià)值。同時(shí),本研究利用實(shí)時(shí)定量PCR檢測(cè)了血漿lncRNAs在經(jīng)歷反復(fù)凍融或室溫放置一段時(shí)間后的穩(wěn)定性,并通過(guò)不同的胃癌細(xì)胞系及胃癌病例術(shù)前術(shù)后的血漿樣本探討血漿lncRNAs可能的來(lái)源;此外,本研究利用細(xì)胞生物學(xué)的方法,初步分析了血漿lncRNAs對(duì)胃癌細(xì)胞惡性表型的影響。結(jié)果:芯片分析篩選出差異表達(dá)的lncRNAs共3,878條,其中在胃癌患者血漿中表達(dá)上調(diào)的有603條,表達(dá)下調(diào)的有3,275條。兩階段血漿樣本驗(yàn)證發(fā)現(xiàn)RP11-47304.5、GUSBP11 和 CTD-2303H24.2 這 3 條 lncRNAs 在胃癌病例血漿中顯著高表達(dá);lncRNAs穩(wěn)定性檢測(cè)發(fā)現(xiàn),在血漿樣本經(jīng)歷反復(fù)凍融或室溫放置一段時(shí)間后,這3條lncRNAs的表達(dá)量均無(wú)顯著差異;ROC曲線分析發(fā)現(xiàn),這3條lncRNAs單個(gè)或者組合對(duì)胃癌診斷均有一定的價(jià)值,其中CTD-2303H24.2單獨(dú)診斷的價(jià)值最大,而這3條lncRNAs的聯(lián)合診斷價(jià)值要高于單個(gè)lncRNA的診斷價(jià)值,AUC為0.818,靈敏度為77.5%,特異度為73.9%;lncRNAs來(lái)源分析發(fā)現(xiàn),不同的胃癌細(xì)胞培養(yǎng)液中均可檢測(cè)到這3條lncRNAs,且胃癌患者術(shù)后血漿中l(wèi)ncRNAs的表達(dá)水平顯著低于手術(shù)前的表達(dá)量;細(xì)胞功能實(shí)驗(yàn)發(fā)現(xiàn),過(guò)表達(dá)RP11-47304.5可促進(jìn)胃癌細(xì)胞的增殖和侵襲能力,干擾RP11-47304.5表達(dá)可抑制胃癌細(xì)胞的增殖和侵襲能力。結(jié)論:本研究發(fā)現(xiàn)了三條新的胃癌相關(guān)的血漿lncRNAs RP11-47304.5、GUSBP11和CTD-2303H24.2,這3條lncRNAs在血漿中穩(wěn)定存在且在胃癌患者血漿中顯著高表達(dá),可以較好地區(qū)分健康對(duì)照和胃癌患者,可作為潛在的胃癌早期診斷的循環(huán)生物標(biāo)志。進(jìn)一步的功能學(xué)研究發(fā)現(xiàn),RP11-47304.5與胃癌細(xì)胞的增殖和侵襲作用相關(guān)。
[Abstract]:Background: our country has been in gastric cancer (gastriccancer, GC) in the country every year, gastric cancer incidence and mortality in the world accounted for about 50% of the total burden of disease, seriously, is the focus of tumor prevention and treatment in patients with gastric cancer. No specific symptoms, the diagnosis most patients have been in advanced stage, the prognosis of advanced gastric cancer is poor. The postoperative five years survival rate is less than 30%, while the good prognosis of early gastric cancer, postoperative five year survival rate can reach more than 90%, so look for to be able to identify early gastric cancer, and Gao Min has the sensitivity and specificity of biomarkers is critical. Long chain non encoding RNAs (long non-coding RNAs, lncRNAs) refers to the length of more than RNA transcription 200 nucleotides and no protein encoding ability. At present, many studies have shown that lncRNAs play an important role in tumor development. In recent years, lncRNAs was found to be released into the circulation and Stable, and the expression level of circulating lncRNAs in cancer patients and healthy controls are significantly different in vivo, may be used as a potential biomarker of tumor and improve the early diagnosis of tumors. The study of circulating lncRNAs is still in the initial stage, and the power source of circulating lncRNA can not clarify the system. Methods: This study used Arraystar Human lncRNA/mRNA V3.0 chip technology, detection and analysis of 5 cases of gastric cancer and 5 cases of sex and age matched healthy controls of the whole genome of plasma lncRNAs in 10 lncRNAs; the most significant expression in the plasma of gastric cancer patients as candidates for lncRNAs in this study, using real-time quantitative PCR (quantitative real-time polymerase chain reaction, qRT-PCR) two phase a large sample of plasma technology (including 50 cases of stage gastric cancer cases and 50 healthy controls plasma plasma, two stage gastric cancer cases including 173 cases of blood Pulp and 173 cases of control plasma) verified by receiver operating characteristic curve (receive operating charactreristic curve, ROC) area under the curve analysis and calculation (area under the curve, AUC), evaluation of a single or a group of lncRNAs on the application value of the diagnosis of gastric cancer. At the same time, this study using real-time quantitative PCR detection of plasma lncRNAs after repeated freeze-thaw stability at room temperature or after a period of time, and through the plasma samples of different gastric cancer cell lines and patients before and after surgery to investigate the source of plasma lncRNAs possible; in addition, this research using the methods of cell biology, preliminary analysis of the effects of plasma lncRNAs on the malignant phenotype of gastric cancer cells. Results: chip analysis showed that 3878 lncRNAs were differentially expressed, the expression in the plasma of patients with gastric cancer in 603, down regulated 3275 stage. The two plasma samples proved to R P11-47304.5, GUSBP11 and CTD-2303H24.2 lncRNAs in 3 gastric cancer cases was significantly higher in plasma expression; lncRNAs stability testing, experience in plasma samples or repeated freezing and thawing at room temperature for a period of time, there were no significant differences in the expression of the 3 lncRNAs; ROC curve analysis shows that the 3 lncRNAs single or combination of have certain value in diagnosis of gastric cancer, the diagnostic value of CTD-2303H24.2 alone, and the diagnostic value of combined diagnostic value of the 3 lncRNAs was higher than that of single lncRNA, AUC was 0.818, the sensitivity was 77.5%, specificity was 73.9%; the lncRNAs source analysis found that the liquid can be detected in the 3 lncRNAs different gastric cancer cell culture that expression level of lncRNAs in plasma and gastric cancer patients was significantly lower than the expression level before surgery; found cell function experiments, overexpression of RP11-47304.5 can promote the proliferation of gastric cancer cells and The invasion ability, interfering the expression of RP11-47304.5 proliferation and invasion inhibition of gastric cancer cells. Conclusion: This study found three new gastric cancer related to plasma lncRNAs RP11-47304.5, GUSBP11 and CTD-2303H24.2, the 3 lncRNAs is stable in plasma in patients with gastric cancer and blood plasma tissues, we can distinguish the healthy control and gastric cancer, early diagnosis of gastric cancer can be used as a circulating biological potential marker. Further functional studies found that RP11-47304.5 cells related to gastric cancer cell proliferation and invasion.

【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R735.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 左婷婷;鄭榮壽;曾紅梅;張思維;陳萬(wàn)青;;中國(guó)胃癌流行病學(xué)現(xiàn)狀[J];中國(guó)腫瘤臨床;2017年01期

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本文編號(hào)：1738493