發(fā)布時(shí)間：2018-05-08 00:40

  本文選題：纖調(diào)蛋白 + 腫瘤血管生成�。� 參考：《第三軍醫(yī)大學(xué)》2016年博士論文

【摘要】：背景腫瘤新生血管為腫瘤細(xì)胞的生長(zhǎng)和轉(zhuǎn)移提供了重要的物質(zhì)來源和結(jié)構(gòu)基礎(chǔ),因此抑制腫瘤血管生成不僅可以切斷腫瘤細(xì)胞的營(yíng)養(yǎng)供給,使腫瘤萎縮退化、缺血壞死,還可以阻斷腫瘤細(xì)胞經(jīng)血管途徑向遠(yuǎn)處轉(zhuǎn)移,抑制轉(zhuǎn)移瘤的形成[10]。小細(xì)胞肺癌(small cell lung cancer,SCLC)是侵襲性極高的惡性腫瘤,SCLC細(xì)胞具有分化程度低、惡性程度高的生物學(xué)特性,易早期侵犯血管發(fā)生遠(yuǎn)處轉(zhuǎn)移。據(jù)統(tǒng)計(jì),廣泛期小細(xì)胞肺癌的平均生存期僅7-12個(gè)月,5年生存率低于2%,疾病預(yù)后極差[3]。另外,小細(xì)胞肺癌也是血管依賴性腫瘤,其豐富的新生血管網(wǎng)為腫瘤細(xì)胞提供氧氣和營(yíng)養(yǎng)物質(zhì),清除局部代謝產(chǎn)物,也為腫瘤細(xì)胞早期局部擴(kuò)散和遠(yuǎn)處轉(zhuǎn)移建立了血性通道。因此,探索引起小細(xì)胞肺癌腫瘤血管異常增生的促血管生成因子及其作用機(jī)制有望為研發(fā)有效的小細(xì)胞肺癌血管生成抑制劑提供研究基礎(chǔ)。纖調(diào)蛋白(fibromodulin,FMOD)是富含亮氨酸重復(fù)序列蛋白聚糖(small leucine-rich proteoglycans,SLRPs)家族成員[11]。近年來諸多研究報(bào)道,FMOD是一種新發(fā)現(xiàn)的內(nèi)源性促血管生成因子,在生理性和病理性血管增生活動(dòng)中具有重要的調(diào)控作用[4-6]。本課題組前期研究比較SCLC親代細(xì)胞株H446及其耐藥細(xì)胞株H446/DDP的基因表達(dá)譜,發(fā)現(xiàn)FMOD在兩種細(xì)胞株中均高表達(dá),兩者mRNA表達(dá)水平?jīng)]有顯著差異。采用RT-PCR和Western Blot進(jìn)一步驗(yàn)證獲得一致結(jié)果。由此說明FMOD的異常表達(dá)與SCLC的發(fā)生發(fā)展存在潛在的聯(lián)系,但與SCLC的獲得性耐藥沒有顯著相關(guān)性。采用免疫組織化學(xué)法(immunohistochemistry,IHC)檢測(cè)FMOD在SCLC患者腫瘤組織標(biāo)本中的表達(dá)情況,發(fā)現(xiàn)FMOD在近一半腫瘤組織標(biāo)本中表達(dá)陽性,而在正常肺組織標(biāo)本中無一例陽性,這進(jìn)一步表明FMOD在SCLC中的異常表達(dá)可能與其發(fā)生發(fā)展密切相關(guān)。基于FMOD的促血管生成作用,我們推測(cè)FMOD可能參與了SCLC的血管生成。目前FMOD在腫瘤血管生成中的作用尚不清楚,其作用機(jī)制也有待深入研究。因此,本課題將在SCLC的疾病研究背景下探討FMOD對(duì)腫瘤血管生成的作用及其分子機(jī)制。首先采用基因沉默技術(shù)下調(diào)FMOD基因表達(dá),接著從臨床水平、動(dòng)物水平、細(xì)胞和分子水平多個(gè)層面研究FMOD對(duì)SCLC腫瘤血管生成的影響,闡明FMOD促SCLC腫瘤血管生成的作用及其相關(guān)機(jī)制。旨在為以FMOD作為靶點(diǎn)開發(fā)新的SCLC抗血管生成藥物提供實(shí)驗(yàn)研究的基礎(chǔ),并最終達(dá)到抑制SCLC發(fā)生發(fā)展,改善疾病預(yù)后的目的。方法1.收集SCLC臨床組織標(biāo)本,以及正常肺泡上皮組織標(biāo)本作為對(duì)照,經(jīng)甲醛固定、石蠟包埋和切片處理后采用免疫組織化學(xué)法檢測(cè)FMOD在組織標(biāo)本中的表達(dá)情況。采集標(biāo)本所對(duì)應(yīng)臨床患者的病歷資料,包括年齡、性別、吸煙、腫瘤分期和預(yù)后等臨床特征參數(shù),采用?2檢驗(yàn)法分析FMOD的表達(dá)水平和各項(xiàng)臨床指針的相關(guān)性。免疫組化法檢測(cè)組織標(biāo)本中血管內(nèi)皮標(biāo)記物CD31的表達(dá),計(jì)數(shù)微血管密度,初步驗(yàn)證FMOD異常表達(dá)與SCLC血管生成的相關(guān)性。2.將靶向FMOD的shRNA序列5’-CCGCATGAAGTACGTCTACTT-3’[7]克隆到慢病毒載體pGLV3/H1/GFP+Puro Vector中構(gòu)建重組慢病毒穿梭質(zhì)粒LV3-shFMOD。同時(shí)構(gòu)建包含無序片段5’-TTCTCCGAACGTGTCACGT-3’的重組慢病毒LV3-shCON。用重組慢病毒轉(zhuǎn)染H446細(xì)胞,嘌呤霉素篩選轉(zhuǎn)染細(xì)胞,Western Blot方法檢測(cè)經(jīng)轉(zhuǎn)染的H446細(xì)胞內(nèi)合成的FMOD和培養(yǎng)液中分泌的FMOD表達(dá)水平,目的是驗(yàn)證H446細(xì)胞是否能合成和分泌FMOD蛋白,LV3-shFMOD是否能有效抑制FMOD的表達(dá)和分泌。3.將重組慢病毒轉(zhuǎn)染后的H446細(xì)胞種植在免疫缺陷小鼠(severe combined immunodeficiency mice,SCID mice)背部皮下組織,建立SCLC移植瘤動(dòng)物模型。定期稱量SCID鼠體重,監(jiān)測(cè)體重隨時(shí)間的變化規(guī)律;定期測(cè)量移植瘤徑長(zhǎng),評(píng)估瘤體的生長(zhǎng)情況;經(jīng)尾靜脈注入對(duì)比增強(qiáng)劑進(jìn)行血管造影B超檢查[9],檢測(cè)腫瘤區(qū)域的血流動(dòng)力學(xué)指標(biāo):血流峰灰階值和峰值時(shí)間,評(píng)估腫瘤的血流灌注情況;摘取移植瘤,用免疫組化法檢測(cè)FMOD和CD31的表達(dá),進(jìn)行微血管計(jì)數(shù),從在體水平進(jìn)一步驗(yàn)證FMOD在腫瘤血管生成中的作用。4.用CCK8試劑盒測(cè)定H446細(xì)胞的增殖活性,transwell小室檢測(cè)H446細(xì)胞的侵襲和遷移能力,分析FMOD對(duì)H446細(xì)胞生物學(xué)活性的影響。收集H446培養(yǎng)上清作為條件培養(yǎng)基(conditioned medium,CM)。將人臍靜脈內(nèi)皮細(xì)胞(human umbilical vein endothelial cells,HUVECs)與CM共培養(yǎng),用CCK8試劑盒測(cè)定不同表達(dá)水平的分泌型FMOD對(duì)HUVECs增殖活性的影響。將HUVECs和H446細(xì)胞分別培養(yǎng)于transwell共培養(yǎng)系統(tǒng)的上室和下室,測(cè)定分泌型FMOD對(duì)HUVECs侵襲和遷移能力的影響。Western Blot檢測(cè)HUVECs與CM共培養(yǎng)后,促血管生成生長(zhǎng)因子的表達(dá)情況。結(jié)果1.共收集27例SCLC肺穿針吸組織標(biāo)本,其中4例臨床病歷資料不全,3例石蠟切片標(biāo)本未檢測(cè)出癌性組織,剩余20例標(biāo)本納入研究,進(jìn)行免疫組化檢測(cè)和統(tǒng)計(jì)學(xué)分析。(1)實(shí)驗(yàn)結(jié)果顯示,FMOD在SCLC臨床組織中的陽性表達(dá)率為55%,陽性表達(dá)定位于細(xì)胞核和細(xì)胞質(zhì),FMOD在正常肺組織中均表達(dá)陰性。采用Fisher’s確切概率法分析得出,FMOD的表達(dá)與患者年齡、性別、吸煙和預(yù)后無相關(guān)性,與SCLC腫瘤分期顯著相關(guān)(P=0.017)。(2)FMOD表達(dá)陽性的組織標(biāo)本微血管數(shù)量為43.89?10.45,FMOD表達(dá)陰性的組織微血管數(shù)量為23.84?10.54,采用t檢驗(yàn)法分析得出,微血管數(shù)量在FMOD陽性表達(dá)的腫瘤組織中顯著高于FMOD陰性組織(P=0.013)。2.重組質(zhì)粒的測(cè)序結(jié)果表明慢病毒重組載體LV3-shFMOD構(gòu)建正確,包含靶向FMOD的shRNA目的序列。慢病毒滴度測(cè)定結(jié)果為1?108 TU/ml,慢病毒感染H446細(xì)胞72h,熒光強(qiáng)度最強(qiáng)。用嘌呤霉素篩選轉(zhuǎn)染成功的細(xì)胞,Western Blot檢測(cè)細(xì)胞內(nèi)和培養(yǎng)上清液中的FMOD表達(dá)水平,結(jié)果顯示轉(zhuǎn)染shFMOD的H446細(xì)胞內(nèi)和培養(yǎng)液中的FMOD含量均顯著性低于對(duì)照組(分別為P0.001,P0.05)。結(jié)果一方面表明H446細(xì)胞能合成和分泌FMOD蛋白;另一方面表明重組質(zhì)粒構(gòu)建正確,shRNA-FMOD能有效抑制FMOD表達(dá)和分泌。3.實(shí)驗(yàn)組(接種H446-shFMOD細(xì)胞)SCID鼠和對(duì)照組(接種H446-shCON細(xì)胞)SCID鼠均在細(xì)胞種植后第4天形成新生瘤體,實(shí)驗(yàn)組平均瘤體體積小于對(duì)照組(P=0.001)。隨后在第7天、第14天、第21天和第28天測(cè)量得出實(shí)驗(yàn)組瘤體體積均小于對(duì)照組(P=0.003,0.003,0.004,0.002)。實(shí)驗(yàn)組SCID鼠的體重在觀察期間持續(xù)增長(zhǎng),對(duì)照組SCID鼠體重在第14天后開始下降(P0.05),惡病質(zhì)表現(xiàn)明顯,出現(xiàn)SCID鼠死亡。血管B超造影檢查發(fā)現(xiàn)實(shí)驗(yàn)組腫瘤區(qū)域的平均血流峰值(10.08?3.49)顯著低于對(duì)照組(26.27?4.93,P0.001),實(shí)驗(yàn)組腫瘤的血流峰值時(shí)間(9.28?4.88s)較對(duì)照組(2.80?0.77s)顯著延長(zhǎng)(P=0.022)。采用免疫組化法對(duì)CD31標(biāo)記的腫瘤微血管染色,計(jì)數(shù)結(jié)果顯示實(shí)驗(yàn)組瘤體組織微血管數(shù)量為16.03?2.84,對(duì)照組相應(yīng)值為35.87?13.41,t檢驗(yàn)分析發(fā)現(xiàn)實(shí)驗(yàn)組微血管數(shù)量明顯減少(P=0.006)。4.H446細(xì)胞轉(zhuǎn)染LV3-shFMOD后,細(xì)胞增殖、侵襲和遷移活性較對(duì)照組均無顯著差異(P0.05)。將HUVECs分別培養(yǎng)于培養(yǎng)上清CM-shFMOD和CM-shCON中,CCK-8檢測(cè)發(fā)現(xiàn)HUVEC在CM-shFMOD中的增殖活性較對(duì)照顯著下降(P0.05);將HUVECs分別與H446-shFMOD和H446-shCON共培養(yǎng),檢測(cè)結(jié)果顯示與H446-shFMOD共培養(yǎng)的HUVECs侵襲和遷移能力均下降,具有統(tǒng)計(jì)學(xué)差異(P0.001)。5.將HUVECs分別培養(yǎng)于條件培養(yǎng)基CM-shFMOD和CM-shCON中,用Western Blot檢測(cè)HUVECs的促血管生成生長(zhǎng)因子的表達(dá)水平。結(jié)果顯示,與對(duì)照組相比,HUVECs與CM-shFMOD共培養(yǎng)后,血管生長(zhǎng)因子VEGF、FGF-2、PDGF-B、TGF-?1表達(dá)均顯著下調(diào)(P0.05)。結(jié)論1.FMOD在SCLC臨床組織中的異常表達(dá)與腫瘤分期相關(guān),提示FMOD可能成為預(yù)測(cè)SCLC腫瘤分期的潛在標(biāo)志物。2.FMOD在SCLC臨床組織中的異常表達(dá)與微血管密度相關(guān),提示FMOD可能參與了小細(xì)胞肺癌的腫瘤血管生成。3.成功構(gòu)建靶向FMOD基因的shRNA重組慢病毒載體,重組載體能顯著抑制SCLC細(xì)胞的FMOD表達(dá)和分泌。4.移植瘤動(dòng)物模型充分證明FMOD能誘導(dǎo)SCLC的腫瘤血管生成,并能增加腫瘤組織的血流灌注,從而促進(jìn)腫瘤的生長(zhǎng)。5.SCLC細(xì)胞合成的內(nèi)源性FMOD對(duì)其自身的增殖活性、侵襲和遷移能力無顯著影響。6.分泌型FMOD促使靜息型內(nèi)皮細(xì)胞的血管生成活性被激活,表現(xiàn)為增殖、侵襲和遷移能力的增強(qiáng)。7.FMOD促腫瘤血管生成的分子機(jī)制是:SCLC細(xì)胞合成并分泌FMOD至細(xì)胞外,經(jīng)旁分泌途徑作用于腫瘤基質(zhì)中的血管內(nèi)皮細(xì)胞。FMOD是多種血管生長(zhǎng)因子的上游調(diào)控子,刺激血管內(nèi)皮細(xì)胞上調(diào)表達(dá)促血管生長(zhǎng)因子VEGF,FGF-2,TGF-?1和PDGF-B,開啟“血管生成開關(guān)”(angiogenic switch),形成有利腫瘤血管生成的局部分子微環(huán)境,從而激活血管內(nèi)皮細(xì)胞的生物學(xué)功能,促進(jìn)腫瘤血管新生。
[Abstract]:Background neovascularization of tumor provides important material source and structural basis for tumor cell growth and metastasis. Therefore, inhibition of tumor angiogenesis can not only cut off the nutritional supply of tumor cells, make tumor atrophy and degeneration, ischemic necrosis, but also block the metastasis of tumor cells through the blood tube pathway to inhibit the formation of metastatic tumor [10 Small cell lung cancer (SCLC) is an aggressive malignant tumor. SCLC cells have low differentiation and high malignant biological characteristics, and early invasion of the distant metastasis of blood vessels. According to the statistics, the average survival time of the extensive stage small cell lung cancer is only 7-12 months, the 5 year survival rate is less than 2%, and the prognosis of the disease is extremely poor [3].. In addition, small cell lung cancer is also a vascular dependent tumor. Its rich neovascularization network provides oxygen and nutrients for tumor cells, scavenging local metabolites, and establishing bloody channels for early local diffusion and distant metastasis of tumor cells. Therefore, it is necessary to explore the angiogenic factors that cause abnormal proliferation of tumor vessels in small cell lung cancer. And its mechanism is expected to provide a basis for research and development of effective angiogenesis inhibitors for small cell lung cancer. Fibromodulin (FMOD) is a member of the small leucine-rich proteoglycans (SLRPs) family, a member of the family [11]., and FMOD is a newly discovered endogenous blood vessel. Generation factors have important regulatory roles in physiological and pathological vascular proliferation activities [4-6].. Earlier studies compared the gene expression profiles of SCLC parent cell line H446 and its drug-resistant cell line H446/DDP. It was found that FMOD was highly expressed in two cell lines, and there was no significant difference in the level of mRNA tables. RT-PCR and Western were used. Blot further verified the consistent results. This shows that the abnormal expression of FMOD is associated with the occurrence and development of SCLC, but has no significant correlation with the acquired resistance of SCLC. The expression of FMOD in the tumor tissue specimens of SCLC patients is detected by immunohistochemistry (immunohistochemistry, IHC), and it is found that FMOD is nearly half of the expression of FMOD. There is no positive expression in the normal lung tissue specimens, which further indicates that the abnormal expression of FMOD in SCLC may be closely related to its development. Based on the angiogenesis effect of FMOD, we speculate that FMOD may be involved in the angiogenesis of SCLC. The role of FMOD in tumor angiogenesis is not yet. It is clear that its mechanism of action needs to be further studied. Therefore, this topic will explore the role and molecular mechanism of FMOD on tumor angiogenesis under the background of SCLC disease research. Firstly, gene silencing technique is used to reduce the expression of FMOD gene, and then the study of FMOD on SCLC tumor from the clinical level, animal level, cell and molecular level. The effect of angiogenesis and the effect of FMOD on the angiogenesis of SCLC tumor and its related mechanism. The aim is to provide the basis for the development of new SCLC anti angiogenesis drugs with FMOD as the target, and to achieve the goal of inhibiting the development of SCLC and improving the prognosis of the disease. The method 1. collects the specimens of SCLC clinical tissue and normal alveolus. As control, the expression of FMOD in tissue specimens was detected by immunohistochemical staining by formaldehyde fixation, paraffin embedding and slice treatment. The medical records of the corresponding clinical patients were collected, including age, sex, smoking, tumor staging and preclinical characteristics, and the analysis of FMOD by the 2 test method. The correlation between expression level and clinical pointer. Immunohistochemical method was used to detect the expression of vascular endothelial marker CD31 in tissue specimens, count microvascular density, and preliminarily verify the correlation between abnormal expression of FMOD and SCLC angiogenesis, and the shRNA sequence 5 '-CCGCATGAAGTACGTCTACTT-3' [7] targeted to FMOD was cloned to the lentivirus vector pGLV3/H1/GFP+. The recombinant lentivirus shuttle plasmid LV3-shFMOD. was constructed in Puro Vector and the recombinant lentivirus containing disordered fragment 5 '-TTCTCCGAACGTGTCACGT-3 was constructed with recombinant lentivirus transfected H446 cells, purinamycin was used to screen transfected cells. Western Blot method was used to detect FMOD in the transfected H446 cells and FM secretion in the culture medium The expression level of OD is to verify whether the H446 cells can synthesize and secrete the FMOD protein, and whether LV3-shFMOD can effectively inhibit the expression of FMOD and secrete.3. to implant the recombinant lentivirus transfected H446 cells into the subcutaneous tissue of the immune deficient mice (severe combined immunodeficiency mice, SCID), and establish a transplanted tumor animal model. The weight of SCID rats was weighed regularly to monitor the change of body weight with time; the length of the tumor size was measured regularly and the growth of the tumor was evaluated; the contrast enhancement agent was injected into the tail vein to perform the angiography B ultrasound examination [9], and the hemodynamic indexes of the tumor area were measured, the peak time and the peak time of the blood flow peak were measured, and the blood flow perfusion of the tumor was evaluated; The expression of FMOD and CD31 was detected by immunohistochemical method, the microvessel count was detected, and the role of FMOD in the angiogenesis of the tumor was further verified from the body level..4. was used to determine the proliferation activity of H446 cells by CCK8 kit, and the ability to detect the invasion and migration of H446 cells by Transwell chamber, and to analyze the effect of FMOD on the biological activity of H446 cells. H446 culture supernatant was collected as a conditioned medium (conditioned medium, CM). Human umbilical vein endothelial cells (human umbilical vein endothelial cells, HUVECs) were co cultured with CM, and the effects of secretory cells with different levels of expression on the proliferation activity were measured with CCK8 kit. The effects of secretory FMOD on the invasion and migration of HUVECs,.Western Blot was used to detect the expression of angiogenic growth factor after co culture of HUVECs and CM. Results 1. a total of 27 specimens of SCLC lung needle aspiration were collected, of which 4 cases of clinical records were incomplete and 3 paraffin sections did not detect cancer. Tissue, the remaining 20 specimens were included in the study to perform immunohistochemical detection and statistical analysis. (1) the results showed that the positive expression rate of FMOD in SCLC clinical tissues was 55%, the positive expression was located in the nucleus and cytoplasm, FMOD was negative in normal lung tissue. The expression of Fisher 's and the expression of FMOD were obtained. There was no correlation between age, sex, smoking and prognosis, and significant correlation with SCLC tumor staging (P=0.017). (2) the number of microvasculature in tissue specimens with positive FMOD expression was 43.89? 10.45, and the number of microvascular tissues with negative FMOD expression was 23.84? 10.54. The number of microvessels was significantly higher in the FMOD positive expression of tumor tissues by t test. The sequencing of FMOD negative tissue (P=0.013).2. recombinant plasmid showed that lentivirus recombinant vector LV3-shFMOD was constructed correctly, including shRNA target sequence targeting FMOD. The result of lentivirus titer was 1? 108 TU/ml, lentivirus infected H446 cells 72h, and the fluorescence intensity was the strongest. The successful transfection of cells was screened with purinamycin, Western Blot detection was fine. The expression level of FMOD in the intracellular and culture supernatant showed that the content of FMOD in H446 cells transfected with shFMOD was significantly lower than that of the control group (P0.001, P0.05). The results showed that H446 cells could synthesize and secrete FMOD protein on the one hand; on the other hand, the construction of recombinant plasmid was correct and shRNA-FMOD could effectively inhibit FMOD. The expression and secretion of.3. experimental group (inoculated H446-shFMOD cells) SCID mice and control group (inoculated H446-shCON cell) SCID mice were formed fourth days after cell cultivation, the average volume of tumor body in the experimental group was less than the control group (P=0.001). Then the volume of the tumor body in the experimental group was less than that of the control group on the seventh day, fourteenth days, twenty-first days and twenty-eighth days (P =0.003,0.003,0.004,0.002). The weight of SCID rats in the experimental group continued to increase during the observation period. The weight of SCID mice in the control group began to decline after fourteenth days (P0.05), and the cachexia was obvious and the SCID mice died. The mean blood flow peak of the tumor area in the experimental group (10.08? 3.49) was significantly lower than that of the control group (26.27? 4.93, P0.001). The peak time of blood flow (9.28? 4.88s) in the experimental group was significantly longer than that of the control group (2.80? 0.77s) (P=0.022). The microvascular staining of CD31 labeled tumor by immunohistochemistry was used in the experimental group. The count results showed that the number of microvessels in the tumor tissue of the experimental group was 16.03? 2.84, the corresponding value of the control group was 35.87? 13.41, and the number of microvessels in the experimental group was detected by t test. After transfecting (P=0.006).4.H446 cells to LV3-shFMOD, the proliferation, invasion and mobility of cells were not significantly different than those of the control group (P0.05). HUVECs was cultured in culture supernatant CM-shFMOD and CM-shCON respectively. CCK-8 detection found that the proliferation activity of HUVEC in CM-shFMOD was significantly lower than that of control (P0.05). H446-shCON co culture, the detection results showed that the invasion and migration ability of HUVECs co cultured with H446-shFMOD decreased, with statistical difference (P0.001).5. HUVECs was cultured in the conditioned medium CM-shFMOD and CM-shCON respectively, and Western Blot was used to detect the expression level of HUVECs vascular generation growth factor. The results showed that the.5. was compared with the control group. After co culture of HUVECs and CM-shFMOD, the expression of vascular growth factor VEGF, FGF-2, PDGF-B, TGF-? 1 was significantly down (P0.05). Conclusion the abnormal expression of 1.FMOD in the SCLC clinical tissue is related to the tumor staging, suggesting that FMOD may be a potential marker for predicting SCLC tumor staging. The abnormal expression of.2.FMOD in clinical tissue and microvascular density may be the potential marker of SCLC tumor staging. Degree related, suggesting that FMOD may be involved in the angiogenesis of small cell lung cancer.3. to successfully construct the recombinant lentivirus vector of the target FMOD gene. The recombinant vector can significantly inhibit the FMOD expression of SCLC cells and secrete the.4. transplanted tumor animal model, which fully proves that FMOD can induce SCLC tumor angiogenesis and can increase the blood flow of the tumor tissue. Perfusion, thus promoting the growth of endogenous FMOD synthesized by.5.SCLC cells, has no significant effect on its own proliferative activity, invasion and migration ability, and the.6. secretory FMOD stimulates the activation of the angiogenesis activity of resting endothelial cells, which shows the molecular mechanism of proliferation, invasion and migration of.7.FMOD to promote tumor angiogenesis. SCLC cells synthesize and secrete FMOD to extracellular, the vascular endothelial cell.FMOD, which is mediated by paracrine pathway in the tumor matrix, is an upstream regulator of a variety of vascular growth factors, stimulating vascular endothelial cells to increase expression of vascular growth factor VEGF, FGF-2, TGF-? 1 and PDGF-B, and to open the "vasculogenic switch" (angiogenic switch). The local molecular microenvironment, which is favorable for tumor angiogenesis, activates the biological functions of vascular endothelial cells and promotes tumor angiogenesis.

【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R734.2

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