【摘要】：隨著環(huán)境污染及食品安全問(wèn)題日趨嚴(yán)重,吸煙、飲酒等不良生活習(xí)慣逐漸向年輕化發(fā)展,致使腫瘤發(fā)病率逐年增高。目前腫瘤已經(jīng)成為威脅人類健康的重大疾病。我國(guó)不管發(fā)病率還是死亡率均居世界前列,給國(guó)民健康造成了極大的危害。腫瘤及其相關(guān)并發(fā)癥的研究越來(lái)越被研究者們所重視。雖然近些年在腫瘤診斷方法及治療手段上已取得長(zhǎng)足的進(jìn)步。但對(duì)于肺癌、肝癌、胰腺癌等惡性腫瘤的治療效果、預(yù)后和遠(yuǎn)期生存率仍然較差。其主要原因歸結(jié)為對(duì)腫瘤發(fā)生發(fā)展機(jī)制的認(rèn)識(shí)不夠深入。脊柱為腫瘤骨轉(zhuǎn)移最為常見(jiàn)的部位,高達(dá)70%的癌癥患者會(huì)發(fā)生脊柱轉(zhuǎn)移。脊柱轉(zhuǎn)移瘤可導(dǎo)致脊髓壓迫及病理性骨折,使患者癱瘓,完全失去自理能力。嚴(yán)重影響患者的生活質(zhì)量,從而加速病情的進(jìn)展。至今國(guó)內(nèi)外學(xué)者對(duì)腫瘤脊柱轉(zhuǎn)移的機(jī)制尚不清楚,致使缺乏有效的治療方法。因此脊柱轉(zhuǎn)移瘤的治療仍然是困擾醫(yī)學(xué)界的一大難題。手術(shù)雖然能有效促進(jìn)恢復(fù),但并發(fā)癥多,創(chuàng)傷較大且短時(shí)間內(nèi)容易復(fù)發(fā),使部分患者在術(shù)后一月內(nèi)死亡。因此,探尋介導(dǎo)腫瘤脊柱轉(zhuǎn)移的關(guān)鍵靶分子,是提高脊柱轉(zhuǎn)移瘤病人臨床療效,改善患者預(yù)后及生存質(zhì)量的新方向。部分腫瘤具有較強(qiáng)的嗜骨性,晚期容易發(fā)生骨轉(zhuǎn)移,例如肝癌、肺癌、乳腺癌等。脊柱為肝癌骨轉(zhuǎn)移最為常見(jiàn)的部位,發(fā)生脊柱轉(zhuǎn)移患者存活率不到3%。肝細(xì)胞癌(hepatocellular carcinoma,HCC)是一種全球范圍內(nèi)常見(jiàn)的惡性腫瘤,占原發(fā)性肝癌的90%以上。患者常常伴有肝內(nèi)及肝外轉(zhuǎn)移。HCC早期臨床癥狀及體征多不典型,很難及早發(fā)現(xiàn)。明確診斷時(shí)大多病程已進(jìn)入中、晚期,失去手術(shù)機(jī)會(huì)。因其惡性程度高、早期發(fā)現(xiàn)難、進(jìn)展速度快、治療復(fù)雜、術(shù)后復(fù)發(fā)及轉(zhuǎn)移率較高等特點(diǎn),使HCC患者療效不佳,預(yù)后較差。以往對(duì)HCC研究主要以microRNA及編碼蛋白的RNA為主,長(zhǎng)鏈非編碼RNA研究相對(duì)較少。目前國(guó)內(nèi)外對(duì)HCC的分子機(jī)制研究還不夠深入,因此我們需進(jìn)一步對(duì)HCC在發(fā)生、發(fā)展及轉(zhuǎn)移過(guò)程中的分子生物學(xué)機(jī)制進(jìn)行探索。尋找可預(yù)測(cè)HCC發(fā)生的分子標(biāo)志物,為HCC診療開辟新途徑。人類基因組是由龐大且復(fù)雜的核苷酸序列排列而成。目前已知可編碼蛋白的基因僅占人類基因組序列的2%,其余98%不具備編碼蛋白能力的序列被認(rèn)為是基因組中無(wú)用的“噪聲”。這類從基因組轉(zhuǎn)錄而來(lái),且不編碼蛋白的RNA分子稱為非編碼RNA(non-coding RNA,ncRNA)。早期,研究者僅僅對(duì)編碼RNA進(jìn)行關(guān)注,但近年來(lái)人們的目光逐漸被非編碼RNA所吸引。隨著研究的不斷深入,發(fā)現(xiàn)長(zhǎng)鏈非編碼RNA(long non-coding RNA,lncRNA)與人類多種疾病如阿爾茨海默癥、腫瘤、亨廷頓氏病的發(fā)生、發(fā)展存在著重要的關(guān)系。LncRNA轉(zhuǎn)錄本長(zhǎng)度大于200nt,且不具備蛋白編碼的能力。最新研究表明,lncRNA在生命活動(dòng)過(guò)程中扮演著重要角色,可通過(guò)調(diào)控mRNA和蛋白表達(dá)水平影響生物學(xué)行為。目前越來(lái)越多的腫瘤相關(guān)lncRNA得以發(fā)現(xiàn),其通過(guò)多種方式調(diào)控著腫瘤的生長(zhǎng)、轉(zhuǎn)移、侵潤(rùn)與凋亡,成為全新腫瘤標(biāo)志物及藥物靶標(biāo),顯著推進(jìn)了腫瘤研究的進(jìn)展。Ji等通過(guò)測(cè)序技術(shù)發(fā)現(xiàn)了肺腺癌轉(zhuǎn)移相關(guān)轉(zhuǎn)錄本(metastasis-associated lung adenocarcinoma transcript 1,MALAT 1),對(duì)225例Ⅲ期非小細(xì)胞肺癌(non-small cell lung cancer,NSCLC)患者進(jìn)行篩查。發(fā)現(xiàn)70例轉(zhuǎn)移患者的樣本中MALAT1過(guò)表達(dá),且具有病程及組織特異性。提示MALAT 1可作為Ⅰ期NSCLC患者潛在標(biāo)志物。該基因在其他腫瘤中也陸續(xù)被發(fā)現(xiàn),但在NSCLC患者中的表達(dá)最為顯著。MALAT 1可通過(guò)激活腫瘤轉(zhuǎn)移相關(guān)基因來(lái)促進(jìn)腫瘤轉(zhuǎn)移,還可參與mRNA前體的可變剪切和絲氨酸/精氨酸剪接因子的磷酸化修飾,來(lái)影響基因的功能及表達(dá)。隨著對(duì)腫瘤基因組學(xué)及轉(zhuǎn)錄組學(xué)研究的不斷深入,研究者發(fā)現(xiàn)不但同種腫瘤中可存在不同的lncRNA,而且同一lncRNA也可在不同腫瘤中差異表達(dá)。HULC最早發(fā)現(xiàn)在肝癌細(xì)胞中過(guò)表達(dá),可作為原發(fā)性肝癌較為敏感的分子標(biāo)志物,其在前列腺癌、胃癌中也出現(xiàn)異常表達(dá)情況。H19作為一個(gè)原癌基因,在多種消化系統(tǒng)腫瘤中表達(dá)量升高。綜上所述,lncRNA在腫瘤學(xué)研究中發(fā)揮著至關(guān)重要的作用。目前,脊柱轉(zhuǎn)移瘤患者多采取微創(chuàng)治療方案,脊柱后路開放式手術(shù)相對(duì)較少。因此組織樣本較為珍貴,收集相對(duì)困難,且骨組織RNA較難提取。致使國(guó)內(nèi)外對(duì)于lncRNA在脊柱轉(zhuǎn)移瘤中的研究尚處空白。LncRNA在HCC中的研究仍然較少,并且絕大多數(shù)lncRNA調(diào)控機(jī)制尚不明確。由此提示我們,是否可以通過(guò)高通量測(cè)序技術(shù)進(jìn)行篩選及驗(yàn)證,找到全新的關(guān)鍵lncRNA作為腫瘤診斷及治療靶標(biāo),為脊柱轉(zhuǎn)移瘤和HCC臨床診療提供新策略�；诖宋覀冮_展了以下三部分研究,現(xiàn)總結(jié)如下。第一章:脊柱轉(zhuǎn)移瘤轉(zhuǎn)錄組lncRNA表達(dá)譜分析目的:建立脊柱轉(zhuǎn)移瘤差異表達(dá)lncRNA及mRNA表達(dá)譜,并對(duì)其長(zhǎng)度及外顯子數(shù)量進(jìn)行預(yù)測(cè)。明確差異表達(dá)mRNA可富集的分子功能、生物學(xué)過(guò)程和疾病通路相關(guān)信息。方法:對(duì)脊柱轉(zhuǎn)移瘤患者配對(duì)組織(腫瘤組織及癌旁組織)中RNA進(jìn)行提取,利用高通量測(cè)序技術(shù)構(gòu)建脊柱轉(zhuǎn)移瘤中差異表達(dá)lncRNA及mRNA表達(dá)譜。隨后通過(guò)pathway分析及GO分析對(duì)差異表達(dá)mRNA的分子功能、參與生物學(xué)過(guò)程及疾病通路進(jìn)行富集。最后對(duì)差異表達(dá)lncRNA長(zhǎng)度及外顯子數(shù)目進(jìn)行預(yù)測(cè),并通過(guò)對(duì)比與編碼基因間位置關(guān)系,將差異表達(dá)lncRNA分類。結(jié)果:通過(guò)高通量測(cè)序技術(shù),在脊柱轉(zhuǎn)移瘤組織中找到171條差異表達(dá)lncRNA和2929條差異表達(dá)mRNA,其中有66條全新未被報(bào)道的lncRNA。對(duì)差異表達(dá)mRNA進(jìn)行pathway分析和GO分析。結(jié)果顯示,脊柱轉(zhuǎn)移瘤中差異表達(dá)的mRNA可富集到31種疾病,79種通路及1535種分子功能,其中包括腫瘤相關(guān)疾病及腫瘤相關(guān)通路。進(jìn)一步明確了脊柱轉(zhuǎn)移瘤中異常表達(dá)mRNA參與的分子功能、生物學(xué)過(guò)程和對(duì)疾病通路的調(diào)節(jié)。171條差異表達(dá)lncRNA中,大部分外顯子數(shù)目小于5個(gè),長(zhǎng)度小于10000nt,均少于mRNA。其中Antisense共有50條,已知的19條,未知的31條;Sence共有53條,已知的51條,未知的2條;Bidirectionsl共有6條,已知的4條,未知的2條;Intergenic共有40條,已知的20條,未知的20條;Intronic共有22條,已知的8條,未知的14條。結(jié)論:成功構(gòu)建了脊柱轉(zhuǎn)移瘤lncRNA、mRNA表達(dá)譜。在171條差異表達(dá)lncRNA中發(fā)現(xiàn)66條全新未被報(bào)道的lncRNA。明確了2929條差異表達(dá)mRNA參與的分子功能、生物學(xué)過(guò)程和對(duì)疾病通路的調(diào)節(jié)。為腫瘤學(xué)研究及后續(xù)實(shí)驗(yàn)提供了寶貴的數(shù)據(jù)資源。第二章:lnc000489在脊柱轉(zhuǎn)移瘤患者血漿中的表達(dá)及臨床特征相關(guān)性分析目的:通過(guò)在血漿中驗(yàn)證,找出脊柱轉(zhuǎn)移瘤患者血漿中高表達(dá)的lncRNA。了解其分子功能、生物學(xué)過(guò)程和疾病通路相關(guān)信息。明確其在脊柱轉(zhuǎn)移瘤中的診斷價(jià)值。方法:從前期構(gòu)建的脊柱轉(zhuǎn)移瘤lncRNA表達(dá)譜中挑選20條全新的lncRNA。將脊柱轉(zhuǎn)移瘤患者血漿設(shè)為實(shí)驗(yàn)組,健康志愿者血漿設(shè)為對(duì)照組。通過(guò)qRT-PCR實(shí)驗(yàn)檢測(cè)候選lncRNA在血漿中表達(dá)量,統(tǒng)計(jì)分析后找出在脊柱轉(zhuǎn)移瘤中具有顯著差異的lncRNA。利用ROC曲線計(jì)算靈敏度和特異性。隨后對(duì)差異表達(dá)lncRNA進(jìn)行共表達(dá)分析,并對(duì)共表達(dá)mRNA進(jìn)行pathway和GO分析。最后統(tǒng)計(jì)分析差異表達(dá)lncRNA與臨床特征間的相關(guān)性。結(jié)果:脊柱轉(zhuǎn)移瘤患者血漿中l(wèi)nc00489表達(dá)量相對(duì)于健康志愿者顯著升高(P0.05),ROC曲線分析顯示lnc00489對(duì)脊柱轉(zhuǎn)移瘤具有一定的診斷價(jià)值。與lnc00489存在共表達(dá)關(guān)系的mRNA共有51條,可富集到實(shí)體腫瘤、卵巢癌、口腔癌、前列腺癌等多種腫瘤相關(guān)疾病和Wnt、Hh、Hedgehog signaling pathway、Basal cell carcinoma等腫瘤相關(guān)通路,進(jìn)一步說(shuō)明了lnc00489可能與腫瘤的發(fā)生、發(fā)展及轉(zhuǎn)移存在相關(guān)性。Lnc00489表達(dá)量與臨床特征進(jìn)行相關(guān)性分析,顯示lnc00489與轉(zhuǎn)移椎體個(gè)數(shù)及是否伴有內(nèi)臟轉(zhuǎn)移顯著相關(guān)。轉(zhuǎn)移椎體數(shù)量越多病情相對(duì)嚴(yán)重。腫瘤主要的轉(zhuǎn)移途徑為血運(yùn)轉(zhuǎn)移,內(nèi)臟及椎體血供相對(duì)豐富。綜上所述lnc00489可能在腫瘤脊柱轉(zhuǎn)移的過(guò)程中發(fā)揮著重要作用,其有望成為潛在的血漿標(biāo)志物,為脊柱轉(zhuǎn)移瘤的診斷及治療提供了新思路。結(jié)論:Lnc00489表達(dá)量在脊柱轉(zhuǎn)移瘤患者血漿中顯著升高,對(duì)脊柱轉(zhuǎn)移瘤具有一定的診斷價(jià)值。與lnc00489共表達(dá)的mRNA可富集到多種腫瘤相關(guān)疾病和通路。血漿中l(wèi)nc00489表達(dá)量與轉(zhuǎn)移椎體數(shù)量和是否伴有內(nèi)臟轉(zhuǎn)移具有相關(guān)性,其有望成為診斷脊柱轉(zhuǎn)移瘤全新的血漿分子標(biāo)志物。第三章:lnc34822在HCC患者血漿中表達(dá)及臨床特征相關(guān)性研究目的:從構(gòu)建的lncRNA數(shù)據(jù)庫(kù)中找到一條在HCC中差異表達(dá)的lncRNA,明確其在HCC中的診斷價(jià)值。方法:利用前期建立的lncRNA數(shù)據(jù)庫(kù),結(jié)合相關(guān)文獻(xiàn)調(diào)研篩選出10條高表達(dá)lncRNA。通過(guò)在HCC患者組織及血漿中驗(yàn)證,明確是否存在高表達(dá)的lncRNA。ROC曲線分析計(jì)算診斷靈敏度和特異性。利用qRT-PCR實(shí)驗(yàn)檢測(cè)候選lncRNA在組織、血漿及細(xì)胞系(Hep3B、HepG-2、SMMC-7721、Huh7.5、LO2)中表達(dá)量。統(tǒng)計(jì)分析差異表達(dá)lncRNA在血漿中的穩(wěn)定性,與外周血細(xì)胞間關(guān)系,不同細(xì)胞系中表達(dá)情況,在HCC患者術(shù)前、術(shù)后及復(fù)發(fā)時(shí)表達(dá)量的變化情況和與臨床特征間的相關(guān)性。結(jié)果:有報(bào)道稱,同種lncRNA可在不同腫瘤中進(jìn)行調(diào)控,同種腫瘤也可受到多種不同lncRNA的影響。結(jié)合文獻(xiàn)調(diào)研從構(gòu)建的脊柱轉(zhuǎn)移瘤lncRNA表達(dá)譜中篩選出10條高表達(dá)lncRNA。經(jīng)過(guò)在HCC患者組織及血漿中驗(yàn)證后,發(fā)現(xiàn)lnc34822在HCC中表達(dá)量顯著上調(diào)(P0.01),并且可以在血漿中穩(wěn)定表達(dá)。Lnc34822在肝癌細(xì)胞系中表達(dá)量明顯高于正常肝細(xì)胞系,且與外周血細(xì)胞沒(méi)有相關(guān)性。ROC曲線分析顯示,lnc34822對(duì)HCC具有一定的診斷價(jià)值。Lnc34822在HCC患者術(shù)后血漿中表達(dá)量明顯降低,而復(fù)發(fā)時(shí)又迅速升高,由此推斷l(xiāng)nc34822有望成為一個(gè)動(dòng)態(tài)監(jiān)測(cè)HCC患者病情進(jìn)展的血漿標(biāo)志物。通過(guò)與臨床信息相關(guān)性分析發(fā)現(xiàn)lnc34822血漿中表達(dá)量與AFP及肝上腫瘤個(gè)數(shù)呈正相關(guān)。肝癌患者血液中AFP含量顯著升高,目前已成為臨床上廣泛應(yīng)用的腫瘤標(biāo)志物。肝上腫瘤數(shù)量越多病情越嚴(yán)重。進(jìn)一步說(shuō)明了lnc34822有望成為HCC潛在的血漿診斷標(biāo)志物。結(jié)論:Lnc34822在HCC患者血漿中穩(wěn)定高表達(dá),且與外周血細(xì)胞沒(méi)有相關(guān)性,對(duì)HCC具有一定的診斷價(jià)值。血漿中l(wèi)nc34822表達(dá)量可動(dòng)態(tài)監(jiān)測(cè)HCC患者病情進(jìn)展,其有望成為診斷HCC全新的血漿分子標(biāo)志物。通過(guò)以上研究,我們成功構(gòu)建了全新lncRNA數(shù)據(jù)庫(kù),為腫瘤學(xué)研究提供了寶貴的數(shù)據(jù)資源。并從中找到與脊柱轉(zhuǎn)移瘤和HCC發(fā)生、發(fā)展相關(guān)的lncRNA,為脊柱轉(zhuǎn)移瘤和HCC臨床診斷及治療提供了新思路。
[Abstract]:With the environmental pollution and food safety becoming more and more serious, bad habits such as smoking and drinking are gradually becoming younger and younger. The incidence of cancer is increasing year by year. Although great progress has been made in the diagnosis and treatment of cancer in recent years, the prognosis and long-term survival rate of lung cancer, liver cancer, pancreatic cancer and other malignant tumors are still poor. Spine is the most common site of bone metastasis. Up to 70% of cancer patients have spinal metastasis. Spinal metastasis can cause spinal cord compression and pathological fracture, paralysis, complete loss of self-care ability. It seriously affects the quality of life of patients, thus accelerating the progress of the disease. The mechanism of spinal metastasis is still unclear, resulting in the lack of effective treatment. Therefore, the treatment of spinal metastasis is still a major problem in the medical community. Although surgery can effectively promote recovery, it has many complications, large trauma and short-term recurrence, so that some patients died within one month after surgery. The key target molecule of spinal metastasis is to improve the clinical efficacy, prognosis and quality of life of patients with spinal metastases. Less than 3%. Hepatocellular carcinoma (HCC) is a common malignant tumor worldwide, accounting for more than 90% of primary liver cancer. Patients often have intrahepatic and extrahepatic metastases. The early clinical symptoms and signs of HCC are often atypical and difficult to detect. Most of the definite diagnosis of HCC has entered the middle, late stage, lost the opportunity for surgery. Because of its high degree of malignancy, difficulty in early detection, rapid progress, complex treatment, high rate of recurrence and metastasis, HCC patients have poor prognosis and poor curative effect. Therefore, we need to further explore the molecular biology mechanism of HCC in the process of occurrence, development and metastasis. To find molecular markers that can predict the occurrence of HCC will open up a new way for the diagnosis and treatment of HCC. Two percent of the sequences, and 98 percent of the other sequences that do not encode proteins, are considered useless "noise" in the genome. RNA molecules transcribed from the genome and that do not encode proteins are called non-coding RNA (ncRNA). In the early days, researchers focused only on coding RNA, but in recent years, people's attention has gradually been focused on non-coding RN. As the research progresses, it has been found that long non-coding RNA (lncRNA) plays an important role in the development of human diseases such as Alzheimer's disease, tumor and Huntington's disease. Nowadays, more and more tumor-associated lncRNAs have been found to regulate the growth, metastasis, invasion and apoptosis of tumors in various ways. They have become new tumor markers and drug targets, and have greatly promoted the progress of tumor research. Ji et al. identified metastasis-associated lung adenocarcinoma transcript 1 (MALAT 1) by sequencing and screened 225 patients with stage III non-small cell lung cancer (NSCLC). These results suggest that MALAT-1 may be a potential marker for stage I NSCLC patients. The gene has been found in other tumors, but it is most significantly expressed in NSCLC patients. MALAT-1 can promote tumor metastasis by activating Tumor Metastasis-related genes, and can also participate in the alterable splicing of mRNA precursors and phosphorylation repair of serine/arginine splicing factors. With the development of tumor genomics and transcriptome, researchers have found that not only different lncRNA can be found in the same tumor, but also the same lncRNA can be differentially expressed in different tumors. HULC was first found to be overexpressed in hepatocellular carcinoma cells, which can be regarded as a more sensitive primary hepatocellular carcinoma. H19, as a proto-oncogene, is highly expressed in a variety of digestive system tumors. In summary, lncRNA plays a vital role in oncology research. At present, spinal metastases are treated with minimally invasive treatment and posterior spinal opening. Therefore, tissue samples are relatively rare, collection is relatively difficult, and it is difficult to extract RNA from bone tissues. As a result, the study of lncRNA in spinal metastases is still blank at home and abroad. Sequencing techniques were screened and validated to identify novel key lncRNA targets for the diagnosis and treatment of spinal metastases and HCC, providing new strategies for clinical diagnosis and treatment. Methods: RNA was extracted from paired tissues (tumor tissues and adjacent tissues) of patients with spinal metastases, and high-throughput sequencing was used to construct spinal rotation. Differentially expressed lncRNA and mRNA expression profiles in tumor metastasis were then analyzed by pathway analysis and GO analysis to enrich the molecular functions of differentially expressed mRNA and participate in biological processes and disease pathways. Results: 171 differentially expressed lncRNA and 2929 differentially expressed mRNAs were found in spinal metastases by high-throughput sequencing, of which 66 were completely new and unreported. Pathway analysis and GO analysis of differentially expressed mRNAs were performed. The results showed that differentially expressed mRNAs in spinal metastases were enriched in 31 diseases and 79 common diseases. Pathway and 1 535 molecular functions, including tumor-related diseases and tumor-related pathways, further clarify the molecular functions, biological processes and regulation of the abnormal expression of mRNA in spinal metastases. There are 50 ntisense, 19 known, 31 unknown; 53 Sence, 51 known, 2 unknown; 6 Bidirectionsl, 4 known, 2 unknown; 40 Intergenic, 20 known, 20 unknown; 22 Intronics, 8 known, 14 unknown. Conclusion: We have successfully constructed lncRNA, mRNA of spinal metastases. Expression profiles. Among 171 differentially expressed lncRNAs, 66 novel unreported lncRNAs were found. The molecular functions, biological processes and regulation of disease pathways of 2929 differentially expressed mRNAs were identified. This provides valuable data for oncology research and follow-up experiments. Chapter 2: The expression of lnc000489 in the plasma of patients with spinal metastases. Objective: To identify the high expression of lncRNA in the plasma of patients with spinal metastases by validating its clinical features. The expression of candidate lncRNA in plasma was detected by qRT-PCR assay. After statistical analysis, significant differences in lncRNA expression in spinal metastases were found. The sensitivity and specificity were calculated by ROC curve. Then, the differential expression of lncR was detected. Results: The expression of lnc00489 in the plasma of patients with spinal metastases was significantly higher than that of healthy volunteers (P 0.05). ROC curve analysis showed that lnc00489 had certain diagnostic value for spinal metastases. There are 51 mRNAs co-expressed with lnc00489, which can be enriched in many tumor-related diseases such as solid tumors, ovarian cancer, oral cancer, prostate cancer and other tumor-related pathways such as Wnt, Hh, Hedgehog signaling pathway, Basal cell carcinoma. This further indicates that lnc00489 may be associated with the occurrence, development and metastasis of tumors. Correlation analysis between Lnc00489 expression and clinical features showed that lnc00489 was significantly associated with the number of metastatic vertebrae and visceral metastasis. Lnc00489 may be a potential plasma marker for the diagnosis and treatment of spinal metastases. Conclusion: The expression of Lnc00489 is significantly elevated in the plasma of patients with spinal metastases and has diagnostic value for spinal metastases. Tumor-related diseases and pathways. The expression of lnc00489 in plasma is correlated with the number of metastatic vertebrae and visceral metastasis. It is expected to be a new plasma molecular marker for the diagnosis of spinal metastases. Chapter 3: Expression of lnc34822 in plasma of HCC patients and its correlation with clinical features. Objective: To find the correlation between the expression of lncRNA in the constructed lncRNA database. Methods: Ten highly expressed lncRNA were screened from the previously established lncRNA database and the related literature. The sensitivity and specificity of diagnosis were determined by validating the presence of highly expressed lncRNA in tissues and plasma of HCC patients. The expression of candidate lncRNA in tissues, plasma and cell lines (Hep3B, HepG-2, SMMC-7721, Huh7.5, LO2) was detected by qRT-PCR. The stability of the differentially expressed lncRNA in plasma, the relationship between the differentially expressed lncRNA and peripheral blood cells, the expression in different cell lines, the changes of the expression in HCC patients before, after and after HCC, and the recurrence of the expression of lncRNA were statistically analyzed. Results: It has been reported that homologous lncRNA can be regulated in different tumors, and homologous tumors can also be affected by a variety of different lncRNA. Ten highly expressed lncRNA were screened from the constructed lncRNA expression profiles of spinal metastases based on literature investigation. After being verified in HCC patients'tissues and plasma, lnc34822 was found. The expression of Lnc34822 in HCC cell line was significantly higher than that in normal liver cell line, and there was no correlation between Lnc34822 and peripheral blood cells. It is concluded that lnc34822 may be a plasma marker for monitoring the progression of HCC.
【學(xué)位授予單位】：中國(guó)人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.7

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8 葛雅麗,鄭敏文,張勁松,楊勇,徐鍵,劉瑩,劉燕麗,趙海濤,常英娟,宦怡;MRI全脊柱移床掃描技術(shù)在脊柱轉(zhuǎn)移瘤中的應(yīng)用[J];放射學(xué)實(shí)踐;2004年11期

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10 陳永順,單國(guó)用,慶明軒,張大勇,張現(xiàn)軍;立體定向放射治療脊柱轉(zhuǎn)移瘤療效分析[J];河南腫瘤學(xué)雜志;2005年05期

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