肿瘤相关纤维母细胞与肿瘤的研究进展

肿瘤的生物学特性不仅是由癌基因和抑癌基因来调控的,还依赖于肿瘤间质等构成的微环境。肿瘤相关纤维母细胞是肿瘤间质中的主要细胞成分,对恶性肿瘤的形成、分化、免疫逃逸和侵袭转移起关键性作用。本文就近年来肿瘤相关纤维母细胞在肿瘤发生、转移中的作用研究进展进行综述。     

肿瘤相关纤维母细胞与肿瘤发生发展的研究进展

肿瘤相关纤维母细胞是肿瘤间质中的主要细胞成分,对恶性肿瘤的形成、分化、免疫逃逸和浸润转移有着重要的调节作用,本文就肿瘤相关纤维母细胞与肿瘤发生发展的研究进展作一综述.     

间质成纤维母细胞与乳腺癌的发生发展研究进展

成纤维母细胞是乳腺癌间质的重要组成成分之一,该细胞与正常乳腺组织间质的成纤维细胞在生物学特性上存在明显的差异,不仅在肿瘤的发生、发展中起重要作用,而且与患者的预后密切相关.近年来,研究还显示乳腺癌相关成纤维母细胞与治疗效果有关,将成为乳腺癌治疗策略中不可忽略的一个方面.     

癌相关纤维母细胞对胃癌细胞增殖、凋亡、迁移和侵袭活性的影响

目的 探讨癌相关纤维母细胞(CAFs)对胃癌细胞增殖、凋亡、迁移和侵袭活性的影响,为胃癌的合理有效治疗提供新的思路和作用靶点。方法 (1)原代培养正常纤维母细胞(NFs)和CAFs,用CCK8实验和流式细胞仪检测间接共培养后,NFs和CAFs对MGC-803细胞增殖和凋亡能力的影响;(2)通过transwell共培养小室建立MGC-803与CAFs相互作用的体外模型,分析CAFs对MGC-803迁移和侵袭能力的影响。结果 (1)与NFs相比,CAFs在体外传代培养过程中,仍高表达波形蛋白(Vimentin)、纤维母细胞活化蛋白(FAP);(2)与NFs组相比,CAFs促进MGC-803细胞增殖、迁移和侵袭,但是抑制其凋亡。结论 CAFs在胃癌的生长、迁移和侵袭等过程中起着重要的作用,并且可能成为胃癌治疗的新靶点。     

肺癌间质纤维母细胞的增殖活性及其意义

背景与目的肿瘤细胞与间质细胞的相互作用在肿瘤的侵袭和转移中起着重要作用。本研究的目的是观察肺癌间质纤维母细胞的增殖活性,探讨纤维母细胞增殖活性与肺癌转移的关系。方法采用免疫组织化学方法检测94例手术切除肺癌组织中纤维母细胞的增殖活性。结果在所有肺癌病例中,无论肿瘤大小,肿瘤中心区瘤细胞PCNA标记指数(LI)均明显低于周围区瘤细胞(P<0.05),而肿瘤中心区纤维母细胞PCNALI显著高于周围区纤维母细胞(P<0.05)。癌旁肺组织中纤维母细胞PCNALI明显低于肿瘤中心区和周围区纤维母细胞(P<0.05)。中心区和周围区纤维母细胞增殖活性与肿瘤细胞的增殖活性明显相关。有淋巴结转移组肿瘤中心区纤维母细胞PCNALI(10.4±6.7)显著高于无转移组(6.5±5.9)(P<0.05),有转移组肿瘤周围区纤维母细胞PCNALI(7.2±6.1)也显著高于无转移组(5.6±4.7)(P<0.05)。结论高度增生瘤细胞和高度增生纤维母细胞在肿瘤组织中的分布有所不同,周围区肿瘤细胞代表瘤细胞的增殖活性,中心区纤维母细胞代表间质纤维母细胞的增殖活性。     

纤维母细胞活化蛋白（FAP—alpha）研究的临床意义

肿瘤间质纤维母细胞，不同于静息状态成人正常组织的纤维母细胞，特征性地表达一种糖蛋白-纤维母细胞活化蛋白（FAPα），FAPα具有二肽酰肽酶及胶原酶蛋白水解活性，因而可以降解二肽，也可以降解凝胶及Ⅰ型胶原，间质降解是肿瘤细胞侵袭的基本条件之一。以FAPα作为肿瘤生物标记以诊断及治疗肿瘤正在成为一个新兴领域。     

纤维母细胞性网状细胞肿瘤的临床病理学特征分析

目的分析纤维母细胞性网状细胞肿瘤的临床病理学特征。方法对1例淋巴结纤维母细胞性网状细胞肿瘤的临床资料进行回顾性分析,同时结合相关文献进行复习。结果患者左侧颈部肿块间歇性增大3年。取左侧颈部淋巴结活检。光镜下淋巴结结构破坏,可见异型肌纤维母细胞样细胞呈条束状、席纹状排列,瘤细胞呈卵圆形、梭形,胞质粉染,边界不清,空泡状核,见1～3个小核仁,核分裂偶见(平均10/10HPF),可见病理性核分裂像。免疫组织化学结果:瘤细胞Vim、α-SMA、Desmin、CD68弥漫强阳性;Calponin、S100弱阳性,Ki-67阳性率约20%,余抗体LCA、CD20、CD3、AE1/AE3、EMA、CD30、CD15、CD1α、HMB45、MelanA、CD34、ALK、CD21和CD35阴性。结论纤维母细胞性网状细胞肿瘤极为罕见,其生物学行为很难判断。临床上需与滤泡及交指状树突细胞肿瘤等鉴别。     

子宫滋养母细胞肿瘤的新类型-上皮样滋养母细胞肿瘤

１概述近年新报告的一种具有癌症特征但与胎盘部位滋养母细胞肿瘤（PSTT）及绒癌不同的滋养母细胞肿瘤，已被Mazur及Kurman犤１犦命名为上皮样滋养母细胞肿瘤（epithelioidtrophoblastictumor，ETT）。以前曾被称为非典型绒癌（atypicalchoriocarcinoma）犤２犦及多发性中间滋养母细胞结节（multiplenodulesofintermediatetrophoblast）犤３犦，迄至２０００年４月共有文献报道犤１～３犦２４例发生于子宫，除１例不详及１例缺乏前次妊娠史外，均在１～１８年分别有足月妊娠、流产、葡萄胎史。Mazur等认为原发于肺者，系因患葡萄胎或绒…     

成纤维母细胞生长因子单克隆抗体的研究

目的 制备并研究bFGF单克隆抗体,以便用于恶性肿瘤的早期诊断。方法 提纯bFGF并用杂交瘤技术获得杂交瘤细胞,再用BALB/C鼠种植收获腹水得到McAb,进行核型、效价、特异性、表位等分析。结果 所得杂交瘤细胞具有双亲细胞染色体特点McAb效价较高,特异性良好,无非特异反应,3种McAb均为IgG型。结论 McAb具有良好的性能,为进一步的临床检测应用奠定了基础。     

肿瘤相关巨噬细胞与肿瘤细胞上皮间质转化关系的研究进展

上皮间质转化(epithelial-mesenchymal transition,EMT)是上皮细胞在形态学上发生向间质细胞表型的转变,在肿瘤的侵袭和转移过程中发挥重要作用,肿瘤相关巨噬细胞(tumor-associatedmacrophage,TAM)是肿瘤的基本成分之一,通过分泌TNF-α、IL-6、EGF、VEGF、MMP-9、uPA等因子诱导肿瘤细胞发生EMT,协同其刺激新生血管形成、降解基质、促进肿瘤细胞发生局部侵袭及远处转移.     

乳腺癌肿瘤相关成纤维细胞对MDA-MB-231细胞增殖、凋亡、黏附及侵袭的影响

目的:研究乳腺癌肿瘤相关成纤维细胞(carcinoma-associated fibroblasts,CAFs)对乳腺癌细胞增殖、凋亡、黏附和侵袭能力的影响.方法:首先,从6例乳腺癌病人肿瘤及癌旁正常乳腺组织块分别分离培养成对的CAFs和正常成纤维细胞(normal fibroblasts,NFs).然后,通过Transwell小室将CAFs或NFs与乳腺癌MDA-MB-231细胞体外共培养,进行增殖(MTT)、侵袭、黏附实验及流式细胞仪细胞凋亡检测.结果:MDA-MB-231与CAFs或NFs共培养后乳腺癌细胞的增殖活性显著增强(P＜0.05).与NFs相比,这种差异在同CAFs共培养组更为明显(P =0.011).与CAFs共培养之后,MDA-MB-231的早期凋亡显著减少(P=0.026);而与NFs共培养之后则无显著差异.CAFs或NFs对MDA-MB-231细胞的细胞周期和晚期凋亡无明显影响.与CAFs或NFs共培养24小时后,MDA-MB-231细胞的侵袭、黏附能力明显增强,黏附数目为(34.7±4.84)个/视野(CAFs)和(20.16±0.09)个/视野(NFs),(P =0.000);侵袭数为(89±4.62)个/视野(CAFs)和(81.6±6.08)个/视野(NFs),(P＜0.05).结论:CAFs能促进乳腺癌MDA-MB-231细胞的增殖、减少其早期凋亡;同时对乳腺癌细胞的黏附和侵袭能力有促进作用.可是NFs的作用较CAFs为弱.有关CAFs影响乳腺癌肿瘤细胞的生物学特性的机制有待于进一步研究.     

The effect of hepatocellular carcinoma-associated fibroblasts on hepatoma vasculogenic mimicry

Vasculogenic Mimicry (VM) is the main source of blood supply in the early stage of tumor growth. Carcinoma-associated fibroblasts (CAFs) are one of the most important host cells in the tumor microenvironment. Some studies have found that CAFs can promote tumor angiogenesis, but there are few reports on the relationship between CAFs and VM. Tissue samples were collected from 60 cases of hepatocellular carcinoma (HCC) and 10 persons with normal liver function. The relationship between VM expression and clinicopathologic features was analyzed. Furthermore, the relationship between VM expression and vimentin or α-SMA expression was analyzed. Primary culture of hepatocellular CAFs and the collection of conditioned media were carried out. The effects of hepatocellular CAF conditioned medium on the formation of VM and the levels of VM-related proteins and genes in MHCC-97H cells were studied. The positive rate of VM was 35.0% in HCC tissues. There was no VM expression in normal liver tissues. VM expression was related to tumor diameter, Edmondson grade, clinical stage, and liver cirrhosis. The expression of vimentin and α-SMA in VM-positive patients was higher than in VM-negative patients. Different concentrations of hepatocellular CAF conditioned medium could promote the formation of VM and increase the expression of VM-related genes and proteins (MMP2 and EphA2) in MHCC-97H cells. The results show that there was a significant correlation between VM formation and the expression of vimentin or α-SMA in HCC tissues. The conditioned medium of hepatocellular CAFs may promote VM formation and the expression of VM-related genes and proteins (MMP2 and EphA2) in hepatoma cell line MHCC-97H.     

不同分化程度口腔鳞癌旁的成纤维细胞增殖活力比较

背景与目的：有关口腔鳞状细胞癌（OSCC）发生发展的机制目前尚未阐明,过去多数研究仅单纯从上皮的角度进行探讨,忽略了问质（宿主）细胞的变异在OSCC发生发展中的重要作用.本研究探讨在不同分化程度OSCC旁的癌相关成纤维细胞（CAFs）其增殖活性是否存在差异.方法：将前期冻存的第三代不同分化程度OSCC旁的CAFs和同部位正常成纤维细胞（NFs）进行复苏并鉴定,测定细胞生长曲线、有丝分裂指数曲线、MTT法,了解各类细胞的增殖活性是否存在差异.重复试验3次,每次检测3孔细胞,将平均值作为最终结果,并用两因素方差分析对检测值进行统计分析.结果：CAFs每天的细胞生长计数值、有丝分裂计数值、MTF检测值均高于NFs,CAFs的生长及存活能力均强于NFs（P＜0.05）,不同分化程度OSCC旁的CAFs增殖活性的差异有显著性（P＜0.05）,分化程度较低的舌癌旁CAFs的增殖活性更强.结论：OSCC与CAFs存在交互作用,CAFs增殖活性的改变可能对OSCC的分化程度产生影响.     

CAFs对乳腺癌细胞生物学特性的影响及机制探讨

目的：通过乳腺癌间质成纤维细胞（ carcinoma-associated fibroblasts,CAFs）与乳腺癌细胞系MDA-MB-231共培养的体外细胞实验及裸鼠接种的在体动物实验,观察CAFs对乳腺癌细胞增殖、凋亡、侵袭和转移活性的影响,并探讨其作用机制。方法：体外实验：分离培养浸润性导管癌组织中CAFs和正常成纤维细胞（ normal fibroblasts,NFs）,然后分别与乳腺癌细胞MDA-MB-231体外共培养,采用MTT法、流式细胞仪、Ma-trigel人工模拟基底膜法分别检测乳腺癌细胞的增殖、凋亡、细胞黏附和侵袭能力。动物在体实验：选择乳腺癌细胞系MDA-MB-231、CAFs和NFs,结合生理盐水（NS）、基质细胞衍生因子-1（SDF-1）及其配体拮抗剂AMD3100,组成不同的组别并接种于裸鼠（共6组）。观察肿瘤的大小,有无淋巴结、肺、肝脏转移。留取血标本及肿瘤组织行SDF-1表达水平的检测。结果：MDA-MB-231与CAFs和NFs共培养后乳腺癌细胞的增殖活性显著增强,其中CAFs的作用较NFs更强（P＝0.011）;CAFs组的黏附能力（34.70±4.84个/视野）明显强于NFs组（20.16±3.09个/视野）,P＝0.000;而CAFs组的侵袭性（89.0±4.62个/视野）也明显强于NFs组（81.6±6.08个/视野,P＝0.045）。CAFs组中的MDA-MB-231的早期凋亡率（2.9±2.4）较NFs组（5.0±4.2）明显降低（P＝0.026）;MDA231﹢CAFs ﹢NS 组的种植肿瘤平均体积最大（9.092±2.662cm3, P＝0.000）;此外,该组共有4只（66.6％）,MDA231﹢NS组有2只（33.3％）存在腋窝淋巴结转移,未见肝肺转移灶。在MDA231﹢CAFs﹢NS组中,血标本 SDF -1值（75.25±16.23pg/ml）、肿瘤组织标本中 SDF -1mRNA值（11.686±8.926）、组织中SDF-1蛋白表达水平（1.006±0.327）均为最高,与其他各组相比均有统计学差异（ P＝0.000）。结论：CAFs可影响乳腺癌肿瘤细胞的生物学特性,具有促进肿瘤细胞增殖,增强其黏附、侵袭及转移能力。其机制可能是通过乳腺癌间质成纤维细胞分泌SDF-1与其特定的受体CXCR4结合这一信号通路来实现的。     

Tangled fibroblasts in tumor-stroma interactions

Fibrogenesis, expressed as “desmoplasia,” is found in many kinds of tumors and considered to be associated with malignancy. In fact, fibroblasts in cancer stroma have attracted considerable attention in the last 2 decades. Although it is accepted that fibroblasts are important in tumor development and progression, the mechanisms involved are highly complex and difficult to disentangle. Which role do fibroblasts play in tumors—are they friends or foes? Can fibroblasts account for the relationship between inflammation, fibrosis and tumor? Do fibroblasts acquire gene changes during carcinogenesis and how many signaling pathways are involved in tumor‐stroma interactions? What is the outcome of fibroblast activation in tumors—senescence, death or recovery to a progenitor cell such as the fibrocyte? These questions still need further exploration and will be discussed in this review.     

Cancer associated fibroblasts: the dark side of the coin

Valid experimental evidence has recently shown that progression of malignant tumors does not depend exclusively on cell-autonomous properties of the cancer cells, but is also deeply influenced by tumor stroma reactivity and undergoes a strict microenvironmental control. Beside structural environmental components as extracellular matrix (ECM) or hypoxia, stromal cells as macrophages, endothelial cells, and cancer-associated fibroblasts (CAFs) play a definite role in cancer progression. This review summarizes our current knowledge on the role of CAFs in tumor progression towards an aggressive phenotype, with particular emphasis on invasiveness, stemness, and preparation of metastatic niche. The controversial origins of CAFs as well as the therapeutical implications of targeting CAFs for anticancer therapy are discussed.     

P53 mutations in stromal fibroblasts sensitize tumors against chemotherapy

The efficacy of chemotherapy is usually viewed as the outcome of cancer-cell-autonomous processes while the contribution of stroma is being overseen. Here we show that p53 mutations in stromal fibroblasts, a genetic lesion that is detectable in primary breast, prostate and probably other cancers, while they accelerate tumorigenesis they also sensitize tumours against conventional chemotherapy by doxorubicin and cis-platinum. The mechanism by which p53 of stromal fibroblasts affects the response of a tumour against chemotherapy is likely to involve the induction of senescence in the fibroblasts which in turns results in the production of growth factors acting onto the cancer cells by paracrine mechanisms. Our findings identify stromal fibroblasts as important modulators of the efficacy of anticancer therapy.     

The role of fibroblasts in the modulation of integrin-dependent interactions between the gastric cell line HGT-1 and fibronectin

Fibronectin plays an important role in gastric cancer progression. However, little is known about the microenvironmental factors modulating integrin-dependent interactions between gastric cancer cells and fibronectin. We therefore studied the regulation by fibroblasts of the integrin-dependent adhesion and migration of the gastric cancer cell line HGT-1 onto fibronectin. We first determined, by immunofluorescence, immunoblotting and flow cytometry, that HGT-1 cells expressed alpha3, alpha5, alpha6, alphaV and beta1 integrin chains, and the alphaVbeta3 and alphaVbeta5 dimers. We verified that HGT-1 cells xenografted to the immunosuppressed newborn rat retained the integrin repertoire detected in vitro and were able to induce the formation of tumors rich in fibronectin. By using an in vitro assay in the presence of neutralizing antibodies, we verified that HGT-1 adhesion and migration onto fibronectin involved beta1, alphaV and alpha5 integrin chains; we verified, by using an in situ adhesion test to rat gastric wall frozen sections, that in situ HGT-1 adhesion to fibronectin was integrin dependent. In coculture experiments, we showed that organ-specific fibroblasts from stomach, lung and dermis were able to induce, in a site-specific manner, the expression of beta1, alpha5 and alphaV integrin chains in HGT-1 cells, their integrin-dependent adhesion and migration on fibronectin and their capacity to secrete oncofetal fibronectin. In conclusion, our results show the capacity for tissue-derived fibroblasts to modulate the integrin-dependent interactions between the gastric cell line HGT-1 and fibronectin. They strongly suggest that, in gastric cancer, stromal fibroblasts contribute to promote fibronectin-mediated local invasion by tumor cells.     

Expression of transforming growth factor-beta 1 and growth in soft agar differentiate prostate carcinoma-associated fibroblasts from normal prostate fibroblasts

Carcinoma-associated fibroblasts (CAF) promote tumor progression of pre-neoplastic epithelial cells. To investigate the basis of this phenomenon, we compared the properties of fibroblasts cultured from normal human prostate (NHPF) to prostate CAF. NHPF and CAF were assayed for growth potential, cell death and proliferative capacity by measuring population doubling time, cell cycle distribution and capability to form colonies in soft agar. Resistance to genotoxic (UV radiation: 0-50 J/cm2) and chemotoxic (0-200 nM Taxol) agents were compared between CAF and NHPF by measuring cell viability and cell cycle analysis. Transforming growth factor beta1 (TGF-beta1) immunoreactivity was assessed in non-malignant and malignant prostatic tissue. No detectable differences were found when comparing CAF and NHPF with respect to population doubling time, cell cycle distribution and response to genotoxic and chemotoxic agents. The mean number of colonies in soft agar was 120.5 for CAF vs. 18.2 for NHPF (p < 0.05). Because TGF-beta1 and matrix metalloproteinase (MMP)-9 have been associated with growth of fibroblasts in soft agar and tumor promotion, we measured the expression of these factors in NHPF and CAF by ELISA. There was no difference in expression of MMP-9; however, TGF-beta1 was expressed in higher concentrations in CAF than in NHPF (p < 0.0014). Furthermore, TGF-beta1 expression was higher in the carcinoma-associated stroma of prostate cancer tissue than stroma of non-malignant prostatic tissue. Increased capability of CAF as compared to NHPF to form colonies in soft agar may be due to a higher expression of TGF-beta1 and correlates with the ability of CAF to promote malignant progression of prostate epithelial cells.