發(fā)布時(shí)間：2018-05-21 14:14

  本文選題：肝細(xì)胞性肝癌 + 肝星狀細(xì)胞��； 參考：《第三軍醫(yī)大學(xué)》2016年博士論文

【摘要】：研究背景肝細(xì)胞性肝癌(Hepatocellular carcinoma,HCC)是一類全球范圍高發(fā)病率和高死亡率的惡性腫瘤。肝星狀細(xì)胞(Hepatic stellate cells,HSCs)已被證實(shí)可以通過促進(jìn)腫瘤免疫抑制來促進(jìn)HCC的進(jìn)展,然而其具體機(jī)制尚不完全清楚。半乳凝集素-1(galectin-1)是半乳凝集素家族的重要成員之一,已被發(fā)現(xiàn)可以通過促進(jìn)免疫抑制來促進(jìn)腫瘤進(jìn)展,在HCC中亦有著顯著的促癌活性,然而其相關(guān)機(jī)制仍有待深入探討。mi RNA-22是mi RNA中的一種,其在HCC中的顯著抑癌作用已經(jīng)得到證實(shí),然而相關(guān)機(jī)制仍不明確。鑒于HSC和galectin-1在HCC中相似的促腫瘤進(jìn)展機(jī)制以及前期研究中發(fā)現(xiàn)的galectin-1和mi RNA-22間可能存在的靶向調(diào)控關(guān)系,我們推測mi RNA-22可能通過調(diào)控HSC中g(shù)alectin-1表達(dá)發(fā)揮其在HCC中的抑癌作用。因此,我們通過研究HSC和galectin-1在促免疫抑制以及促HCC腫瘤進(jìn)展作用中的相關(guān)性,并尋找其與mi RNA-22靶向調(diào)控的相關(guān)證據(jù),為探索以mi RNA-22-HSC-galectin-1為靶點(diǎn)的HCC治療提供新的理論基礎(chǔ)和實(shí)驗(yàn)依據(jù)。研究方法和結(jié)果1、半乳凝集素-1在人肝星狀細(xì)胞中的表達(dá)及作用研究從西南醫(yī)院肝膽外科收集肝臟局部良性疾病手術(shù)患者正常肝臟組織,提取人原代肝星狀細(xì)胞并鑒定。采用與CD3+T淋巴細(xì)胞共培養(yǎng)的方式觀察到HSC擁有促T細(xì)胞凋亡以及促Th1/Th2免疫平衡偏移的能力。采用RT-PCR、Western blot、免疫組化以及ELISA等方式證實(shí)人HSC中有g(shù)alectin-1的表達(dá)和分泌。構(gòu)建galectin-1抑制和過表達(dá)載體,干預(yù)HSC中g(shù)alectin-1表達(dá)后發(fā)現(xiàn)其促T細(xì)胞凋亡以及促Th1/Th2免疫平衡偏移的能力與其galectin-1表達(dá)情況呈正向相關(guān)。以上結(jié)果證實(shí)(1)活化人肝星狀細(xì)胞中確有g(shù)alectin-1的表達(dá)和分泌;(2)活化人肝星狀細(xì)胞可以促CD3+T淋巴細(xì)胞凋亡及Th1/Th2免疫平衡偏移,且此能力與其galectin-1表達(dá)密切相關(guān)。2、半乳凝集素-1在肝星狀細(xì)胞中的表達(dá)與肝細(xì)胞性肝癌腫瘤進(jìn)程的相關(guān)性研究從西南醫(yī)院肝膽外科收集肝細(xì)胞性肝癌手術(shù)患者腫瘤肝組織,提取人原代肝星狀細(xì)胞。比較不同來源HSC(HCC源:Ca-HSCs,正常肝組織源:N-HSCs)中g(shù)alectin-1表達(dá)及免疫調(diào)節(jié)能力差異,發(fā)現(xiàn)Ca-HSCs擁有明顯更高的galectin-1表達(dá)以及更強(qiáng)的促T細(xì)胞凋亡和促Th1/Th2免疫平衡偏移能力。收集臨床HCC患者腫瘤組織樣本和病例資料,通過免疫熒光染色證實(shí)了活化HSC標(biāo)志物α-SMA和galectin-1在肝組織中的共表達(dá)情況,通過連續(xù)石蠟切片免疫組化染色發(fā)現(xiàn)了活化HSC標(biāo)志物α-SMA和galectin-1在HCC組織間質(zhì)中的高度共表達(dá)情況。通過統(tǒng)計(jì)學(xué)分析發(fā)現(xiàn)了galectin-1表達(dá)以及CD3表達(dá)與HCC患者臨床病理特征參數(shù)的顯著相關(guān)性,進(jìn)一步通過生存分析發(fā)現(xiàn)galectin-1高表達(dá)以及CD3低表達(dá)都是預(yù)示HCC患者不良預(yù)后的獨(dú)立危險(xiǎn)因素,且這兩者間呈明顯負(fù)性相關(guān)。以上結(jié)果表明,肝星狀細(xì)胞可以通過galectin-1促進(jìn)肝細(xì)胞性肝癌中免疫抑制環(huán)境的形成并促進(jìn)腫瘤進(jìn)展。3、mi RNA-22與肝星狀細(xì)胞中半乳凝集素-1表達(dá)及促免疫抑制功能的相關(guān)性研究通過RT-PCR檢測不同組織源肝星狀細(xì)胞中mi RNA-22的表達(dá)情況后發(fā)現(xiàn),在HSC中mi RNA-22的表達(dá)與galectin-1表達(dá)呈顯著負(fù)相關(guān),且癌組織源HSC中mi RNA-22表達(dá)明顯低于正常肝組織源HSC。通過雙熒光素酶實(shí)驗(yàn)證實(shí)了mi RNA-22與galectin-1的靶向調(diào)控關(guān)系。通過mi RNA-22 mimic轉(zhuǎn)染HSC發(fā)現(xiàn),HSC中mi RNA-22的表達(dá)升高可以顯著下調(diào)其galectin-1表達(dá),并抑制HSC促免疫抑制功能的發(fā)揮。以上結(jié)果表明,mi RNA-22可以抑制肝星狀細(xì)胞中g(shù)alectin-1的表達(dá)以及其參與誘導(dǎo)的促T細(xì)胞凋亡作用和促Th1/Th2免疫平衡偏移作用,而這種負(fù)性調(diào)控可能還影響了肝星狀細(xì)胞在肝細(xì)胞性肝癌中的促免疫抑制及促腫瘤進(jìn)展作用。結(jié)論肝星狀細(xì)胞可以通過galectin-1促進(jìn)肝細(xì)胞性肝癌中免疫抑制環(huán)境的形成進(jìn)而促進(jìn)腫瘤進(jìn)展,而這一作用機(jī)制受到mi RNA-22的抑制和調(diào)控。
[Abstract]:Hepatocellular carcinoma (HCC) is a kind of malignant tumor with high global incidence and high mortality. Hepatic stellate cells (HSCs) has been proved to promote the progression of HCC by promoting tumor immunosuppression, but its specific mechanism is not completely clear. Galactin -1 (Gal) -1 (Gal). Ectin-1) is one of the most important members of the galactoglutpins family, which has been found to promote tumor progression by promoting immunosuppression and also has significant cancer promoting activity in HCC. However, its mechanism remains to be discussed in depth of.Mi RNA-22 as one of MI RNA, and its significant tumor suppressor effect in HCC has been confirmed, however, related machines The system is still unclear. In view of the similar tumor promoting mechanism of HSC and galectin-1 in HCC and the possible targeting regulation relationship between galectin-1 and MI RNA-22 found in previous studies, we speculate that MI RNA-22 may play its role in inhibiting cancer by regulating galectin-1 expression in HSC. The correlation of n-1 in promoting immunosuppression and promoting the progression of HCC tumor, and looking for evidence related to the targeting regulation of MI RNA-22, providing a new theoretical basis and experimental basis for exploring HCC therapy targeting mi RNA-22-HSC-galectin-1. Methods and results 1, the expression and role of galactocoagulin -1 in human hepatic stellate cells The primary hepatic stellate cells were extracted from the patients with benign liver diseases in the Department of hepatobiliary surgery of Southwest Hospital, and the human primary hepatic stellate cells were extracted and identified. The ability of HSC to promote T cell apoptosis and promote Th1/Th2 immune balance migration was observed by co culture with CD3+T lymphocyte. RT-PCR, Western blot, immunohistochemistry were used. The expression and secretion of galectin-1 in human HSC were confirmed by ELISA and other methods. The expression of galectin-1 inhibition and overexpression vector was constructed. After the intervention of galectin-1 expression in HSC, the ability to promote the apoptosis of T cells and the ability to promote the migration of Th1/Th2 immune balance were positively correlated with the galectin-1 expression. The results confirmed (1) the activation of human hepatic stellate cells was confirmed. The expression and secretion of galectin-1; (2) activation of human hepatic stellate cells can promote CD3+T lymphocyte apoptosis and Th1/Th2 immune balance migration, and this ability is closely related to the expression of galectin-1 and.2. The relationship between the expression of galactoagglutinin -1 in hepatic stellate cells and the progression of hepatocellular carcinoma in the Department of hepatobiliary surgery is collected from the Southwest Hospital The human primary hepatic stellate cells were extracted from the liver tissues of the patients with hepatoma hepatoma, and the expression of galectin-1 and the immune regulation were compared in different sources of HSC (HCC source: Ca-HSCs, normal liver tissue source: N-HSCs). It was found that Ca-HSCs had a significantly higher galectin-1 expression and stronger T cell apoptosis and Th1/Th2 immune balance bias. The tumor tissue samples and case data of HCC patients were collected and the co expression of the activated HSC marker alpha -SMA and galectin-1 in the liver tissues was confirmed by immunofluorescence staining. The high co expression of the activated HSC marker alpha -SMA and galectin-1 in the interstitial tissue of HCC was found by continuous paraffin immunohistochemical staining. The significant correlation between the expression of galectin-1 and the expression of CD3 and the clinicopathological parameters of HCC patients was found by statistical analysis. Further through survival analysis, the high expression of galectin-1 and the low expression of CD3 were independent risk factors for the poor prognosis of HCC patients, and there was a significant negative correlation between these two factors. It is clear that hepatic stellate cells can promote the formation of immunosuppressive environment in hepatocellular carcinoma by galectin-1 and promote tumor progression.3. The correlation between the expression of MI RNA-22 and the expression of galactoagglutinin -1 in hepatic stellate cells and the function of promoting immunosuppression by RT-PCR detection of the expression of MI RNA-22 in the hepatic stellate cells of different tissues by RT-PCR The expression of MI RNA-22 in HSC was significantly negatively correlated with the expression of galectin-1, and the expression of MI RNA-22 in HSC of cancer tissue was significantly lower than that of normal liver tissue. The relationship between MI RNA-22 and galectin-1 was confirmed by the experiment of HSC. through the double luciferase experiment. The results showed that MI RNA-22 could inhibit the expression of galectin-1 in hepatic stellate cells and the effect of its involvement in inducing apoptosis of T cells and promoting the migration of Th1/Th2 immune balance in hepatic stellate cells, and this negative regulation may also affect hepatic stellate cells in liver cell liver. The results suggest that MI RNA-22 can inhibit the expression of galectin-1 in hepatic stellate cells. Conclusion hepatic stellate cells can promote the formation of immunosuppressive environment in hepatocellular carcinoma and promote tumor progression through galectin-1, and the mechanism of this action is inhibited and regulated by Mi RNA-22.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R735.7

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 ;Circulating galectin-3 in the bloodstream:An emerging promoter of cancer metastasis[J];World Journal of Gastrointestinal Oncology;2010年04期

2 楊婷婷;程若川;;galectin-3與幾種甲狀腺疾病的關(guān)系[J];中國普外基礎(chǔ)與臨床雜志;2011年08期

3 María L Bacigalupo;Malena Manzi;Gabriel A Rabinovich;María F Troncoso;;Hierarchical and selective roles of galectins in hepatocarcinogenesis, liver fibrosis and inflammation of hepatocellular carcinoma[J];World Journal of Gastroenterology;2013年47期

4 鄭華川;王威;關(guān)一夫;平尷}一;;CCl_4誘發(fā)galectin-3基因敲除鼠急性肝損傷的實(shí)驗(yàn)研究[J];中國醫(yī)科大學(xué)學(xué)報(bào);2008年05期

5 童華生;張亞歷;姜泊;蘇磊;;galectin-1表達(dá)下調(diào)對LST-R1細(xì)胞侵襲表型的影響[J];胃腸病學(xué);2010年04期

6 張巖;佟盛春;王永來;趙潤秋;;galectin-3在子宮內(nèi)膜病變中表達(dá)的研究[J];中國醫(yī)科大學(xué)學(xué)報(bào);2008年05期

7 ;Pancreatic stellate cells promote proliferation and invasiveness of human pancreatic cancer cells via galectin-3[J];World Journal of Gastroenterology;2008年13期

8 童華生;張亞歷;姜泊;蘇磊;;RNAi沉默galectin-1表達(dá)對黏附LST-R1細(xì)胞生長的影響[J];胃腸病學(xué)和肝病學(xué)雜志;2009年12期

9 張延美,董娟,胡克霞;galectin-3與甲狀腺腫瘤[J];山西醫(yī)科大學(xué)學(xué)報(bào);2003年03期

10 張磊;張思瑾;常明;胡林森;;蛋白酶體抑制劑誘導(dǎo)的帕金森病模型中g(shù)alectin-1含量變化的研究[J];腦與神經(jīng)疾病雜志;2010年02期

相關(guān)會議論文 前10條

1 ;A novel fungal galectin induces apoptosis through inhibition of farnesyltransferase activity[A];湖北省生物工程學(xué)會2004年年會學(xué)術(shù)報(bào)告及論文摘要匯編[C];2004年

2 ;Galectin-3 and wound repair[A];第七屆全國創(chuàng)傷學(xué)術(shù)會議暨2009海峽兩岸創(chuàng)傷醫(yī)學(xué)論壇論文匯編[C];2009年

3 黃健;金潔;;骨髓增生異常綜合征患者galectin-3和mdr-1表達(dá)及其相關(guān)性研究[A];2012年浙江省血液病學(xué)年會論文集[C];2012年

4 張亞歷;賴卓勝;姜泊;童華生;;RNAi沉默galectin-1表達(dá)對LST-R1細(xì)胞惡性表型的影響[A];中華醫(yī)學(xué)會第七次全國消化病學(xué)術(shù)會議論文匯編（下冊）[C];2007年

5 童華生;張亞歷;姜泊;;RNAi沉默galectin-1表達(dá)對LST-R1細(xì)胞表型的影響[A];2009年全國危重病急救醫(yī)學(xué)學(xué)術(shù)會議論文匯編[C];2009年

6 張亞歷;姜泊;童華生;;RNAi沉默galectin-1表達(dá)對LST-R1細(xì)胞增殖與凋亡的影響[A];中華醫(yī)學(xué)會第七次全國消化病學(xué)術(shù)會議論文匯編（下冊）[C];2007年

7 Yufu Ye;Shen Yan;Xiaojun Xie;Shusen Zheng;;Galectin-1 Attenuates Hepatic Ischemia Reperfusion Injury in Mice through an IL-10-dependent Mechanism[A];2013中國器官移植大會論文匯編[C];2013年

8 Yufu Ye;Sheng Yan;Qiyi Zhang;Yuan Ding;Fei Sun;Yang Tian;Lin Zhou;Haiyang Xie;Shusen Zheng;;Galectin-1 Attenuates Hepatic Ischemia Reperfusion Injury in Mice through an IL-10-dependent Mechanism[A];2013年浙江省腸外腸內(nèi)營養(yǎng)學(xué)學(xué)術(shù)年會論文匯編[C];2013年

9 童華生;賴曉嶸;張亞歷;姜泊;;Galectin-1對LoVo細(xì)胞生長惡性表型的影響[A];中華醫(yī)學(xué)會第七次全國消化病學(xué)術(shù)會議論文匯編（下冊）[C];2007年

10 童華生;賴曉嶸;張亞歷;姜泊;賴卓勝;;細(xì)胞膜表面Galectin-1抗體阻斷對LST-R1細(xì)胞與Ⅰ型膠原黏附的影響[A];中華醫(yī)學(xué)會第七次全國消化病學(xué)術(shù)會議論文匯編（下冊）[C];2007年

相關(guān)博士學(xué)位論文 前8條

1 鄭玲燕;Galectin-1介導(dǎo)NFs向CAFs轉(zhuǎn)化與胃癌apatinib敏感性的關(guān)系的研究[D];浙江大學(xué);2016年

2 張濤;酸堿修飾的檸檬果膠的分離純化及其與半乳凝素-3相互作用的構(gòu)效關(guān)系研究[D];東北師范大學(xué);2016年

3 游逾;miRNA-22抑制肝星狀細(xì)胞中g(shù)alectin-1表達(dá)及其促肝細(xì)胞性肝癌免疫抑制作用的研究[D];第三軍醫(yī)大學(xué);2016年

4 張鴻;galectin-3在CCl_4肝損傷中作用機(jī)制的研究[D];中國醫(yī)科大學(xué);2005年

5 童華生;大腸側(cè)向發(fā)育型腫瘤差異蛋白篩選及galectin-1對大腸癌細(xì)胞表型的影響[D];第一軍醫(yī)大學(xué);2006年

6 高錫強(qiáng);CD45糖基化對galectin-3結(jié)合與凋亡誘導(dǎo)的影響&感染途徑對艾滋病恒河猴CD4~+T細(xì)胞表達(dá)CD95的影響[D];北京協(xié)和醫(yī)學(xué)院;2013年

7 梁美艷;半乳糖結(jié)合蛋白（Galectin）-9抗腫瘤作用及其在子宮頸癌組織中的表達(dá)和臨床意義[D];中國醫(yī)科大學(xué);2008年

8 張榮成;蛋白標(biāo)記物在心力衰竭患者的臨床應(yīng)用[D];北京協(xié)和醫(yī)學(xué)院;2015年

相關(guān)碩士學(xué)位論文 前10條

1 顧彩紅;Tim3及其配體galectin-9在急性髓系白血病的表達(dá)及功能研究[D];蘇州大學(xué);2016年

2 張國強(qiáng);肝細(xì)胞肝癌中g(shù)alectin-9表達(dá)及其臨床意義[D];南方醫(yī)科大學(xué);2011年

3 孫婭莉;仔豬galectins基因的組織分布及斷乳前后其在胃腸道表達(dá)差異研究[D];河南農(nóng)業(yè)大學(xué);2010年

4 劉守勝;牙鲆半乳糖凝集素galectin-1的抗病毒活性研究[D];中國海洋大學(xué);2014年

5 雒真龍;Galectin-7在小鼠移植皮膚中的表達(dá)及其意義[D];華中科技大學(xué);2010年

6 郭一川;Skp2、p27、galectin-3在子宮頸上皮內(nèi)瘤變中的表達(dá)及意義[D];新疆醫(yī)科大學(xué);2008年

7 趙世彬;IVF失敗者子宮內(nèi)膜galectin-3表達(dá)及其容受性評估[D];河北醫(yī)科大學(xué);2011年

8 褚芬芬;NLRP3在高糖誘導(dǎo)HK-2細(xì)胞轉(zhuǎn)分化中的作用及其表達(dá)與galectin-1的關(guān)系[D];中南大學(xué);2014年

9 黃青;siRNA干擾大腸癌細(xì)胞株galectin-3蛋白表達(dá)的研究[D];廣州醫(yī)科大學(xué);2013年

10 徐躍;中華蟾蜍與日本蟾蜍皮膚galectin-3 cDNA的克隆、序列分析及原核表達(dá)[D];浙江農(nóng)林大學(xué);2012年

，

本文編號：1919517