内皮素-1促肿瘤血管新生机制

内皮素(ET)是一种由21个氨基酸组成的生物活性肽,有3种异构体(ET-1,ET-2,ET-3),目前研究最多也最清楚的是ET-1,在正常组织中它通过与其受体(ETR)发挥强大的缩血管及促有丝分裂作用.近年的研究证实,ET-1及ETR在多种肿瘤组织中分泌和表达,并起到促进肿瘤组织血管新生的作用.     

过表达HIF-1α和VEGF蛋白与垂体腺瘤侵袭行为的关系

目的:探讨缺氧诱导因子-1α（HIF-1α）和血管内皮生长因子(VEGF)在垂体腺瘤中的表达及与垂体腺瘤侵袭性的关系。方法:收集的32例垂体腺瘤标本中侵袭性垂体腺瘤20例,非侵袭性垂体腺瘤12例,应用免疫组化染色检测HIF-1α和VEGF蛋白的表达。并对侵袭组与非侵袭组的HIF-1α和VEGF蛋白表达及不同大小肿瘤HIF-1α和VEGF蛋白表达进行分析比较。结果:非侵袭组垂体腺瘤中HIF-1α和VEGF蛋白表达阳性率与侵袭组比较,差异有统计学意义（P＜0.05）;垂体腺瘤直径大于30 mm者HIF-1α和VEGF蛋白表达阳性率与直径10~30 mm者比较,差异有统计学意义（P＜0.05）。结论:HIF-1α和VEGF蛋白在侵袭性垂体腺瘤中的表达明显增加,并且与垂体腺瘤侵袭行为密切相关。     

水蛭抑制肿瘤血管生成的作用及其机制

目的 探讨水蛭能否通过改善肿瘤细胞缺氧微环境来抑制肿瘤血管生成及其相关机制.方法 以氯化钴模拟化学缺氧,采用MTT法观察水蛭含药血清对EA.hy926细胞的增殖抑制作用;采用RTPCR法检测缺氧诱导因子-1α(HIF-1α)mRNA、血管内皮生长因子(VEGF) mRNA的表达;Western blot法检测HIF-1α蛋白水平;Matrigel基质胶模拟体外小管形成实验检测小管生成情况.结果 在缺氧条件下,水蛭含药血清能抑制EA.hy926细胞的增殖,且呈时间和剂量依耐性.水蛭含药血清在低氧时可显著抑制HIF-1α mRNA、VEGF mRNA的表达和HIF-1α蛋白水平,与阴性对照组比较,差异有统计学意义(P＜0.05);水蛭高剂量组和阳性对照组HIF 1α mRNA表达及其蛋白表达水平相当(P＞0.05),而VEGF mRNA在高剂量水蛭作用后的表达更少(P＜0.05).小管形成实验中,水蛭含药血清作用后小管数目与阴性对照组相比明显减少,差异有统计学意义(P＜0.05),与阳性对照组相比,差异无统计学意义(P＞0.05).结论 水蛭能通过改善肿瘤缺氧微环境抑制肿瘤血管生成来发挥抗肿瘤作用,其机制可能是通过降低HIF-1α蛋白水平和mRNA的表达,以及降低由HIF-1α所介导的靶基因VEGF mRNA的表达来实现的.     

大黄素抑制人肝癌HepG2细胞血管生成作用及其机制研究

目的 大黄素可促进人肝癌HepG2细胞的凋亡,然而其是否可抑制肝细胞癌(hepatocellular carcinoma,HCC)血管生成及其机制罕见报道.本研究探讨大黄素对人肝癌HepG2细胞血管生成以及缺氧诱导因子-1α (hypoxia inducible factor-1a,HIF-1d)和血管内皮生长因子(vascular endothelial growth factor,VEGF)的影响,以及大黄素体内抗肝癌机制.方法 采用体内鸡胚绒毛尿囊膜(chick chorioallantoic membrane,CAM)实验,实验随机分为阴性对照组(生理盐水)、大黄素低剂量组(10 μmol/L)、大黄素高剂量(20 μmol/L)和阳性对照组(0.15 mg/mL地塞米松),每组各10只,每24 h每组追加同样等量药物,共72 h,观察大黄素对CAM血管生成的抑制作用.体外培养HepG2细胞,以氯化钴(CoCl2)模拟化学缺氧,设立缺氧未处理组[阴性对照组(生理盐水)]、缺氧大黄素低剂量组(10 μmol/L)、缺氧大黄素高剂量组(20 μmol/L)和缺氧阳性对照组[10 μmol/L 5-氟尿嘧啶(5-fluorouracil,5-FU)],每组设6个复孔,处理24 h.采用小管形成实验,观察大黄素对HepG2细胞相对小管数目的影响,免疫细胞化学分析检测HIF-1α和VEGF阳性细胞的表达,Real-Time PCR分析检测HIF-1α mRNA和VEGF mRNA的表达.采用Graphpad 5.0软件进行描述性统计,组间比较采用one-way-ANOWA和Bonferroni检验.结果 CAM实验显示,实验组新生血管数目明显减少,与阴性照组比较,差异有统计学意义,F=13.374,P=0.002.阴性对照组新生血管数目为(31.47±1.81)个.给药后CAM血管生成减少,大黄素低剂量组、高剂量组和地塞米松组分别为(19.48±0.66)、(10.33±1.04)和(9.89±0.57)个,与阴性对照组比较,差异有统计学意义,P值分别为0.041、0.004和0.003;大黄素低剂量组和高剂量组与地塞米松组比较,差异无统计学意义,P值分别为0.539和1.000.小管形成实验显示,实验组相对小管数目明显减少,与阴性照组比较,差异有统计学意义,F=21.529,P＜0.001.阴性对照组相对小管数目为(100.00±0.00)％.给药后,相对小管数目明显减少,大黄素低剂量、高剂量组和阳性对照组分别为(65.85±12.67)％、(46.94±8.34)％和(41.77±7.53)％,与阴性对照组比较,差异有统计学意义,P值分别为0.049、0.001和＜0.001;大黄素低剂量组和高级剂量组与阳性对照组比较,差异无统计学意义,P值分别为0.104和0.069.免疫细胞化学分析显示,大黄素低剂量组、高剂量组和阳性对照组HIF-1α和VEGF阳性细胞显著减少.Real-Time PCR分析显示,实验组HIF-1α mRNA和VEGF mRNA表达降低,与阴性照组比较,差异有统计学意义(F=31.908,P＜0.001;F=25.146,P＜0.001).阴性对照组HepG2细胞HIF-1α mRNA和VEGF mRNA相对表达分别为0.92±0.15和0.95±0.15;大黄素低剂量组HIF-1α mRNA和VEGF mRNA相对表达分别为0.45±0.07和0.51±0.08,与阴性对照组比较,差异有统计学意义,P值分别为0.021和0.013;大黄素高剂量组HIF-1α mRNA和VEGF mRNA相对表达分别为0.32±0.05和0.40±0.07,与阴性对照组比较,差异有统计学意义,P值分别为＜0.001和0.001;阳性对照组HIF-1α mRNA和VEGF mRNA相对表达分别为0.30±0.04和0.34±0.05,与阴性对照组比较,差异有统计学意义,P值均＜0.001;大黄素低剂量组、高剂量组HIF-1α mRNA和VEGF mRNA相对表达与阳性对照组比较,差异无统计学意义,P值分别为0.663、0.362、0.443和1.000.结论 大黄素可能通过抑制HIF-1d mRNA的表达,从而降低VEGF mRNA的表达来抑制HepG2细胞新生血管的形成.     

肺癌中HIF-1α、VEGF表达及其与放射治疗关系的研究进展

为解决非小细胞肺癌放射治疗后期肿瘤乏氧而产生放射耐受,各国学者尝试使用乏氧细胞放疗增敏剂、高压氧疗、中子疗法以及放疗前辅助化疗等方法,但疗效无明显提高。靶向药物的出现看似能解决一定的问题,但往往对自身机体有一定的损伤,并且有同时耐受的现象发生,如何解决这些问题,是肿瘤放化疗研究热点。研究发现,肺癌患者HIF-1α及VEGF蛋白表达情况对于放射治疗中应用增敏剂及治疗方案的制定有一定的意义。文章主要对此综述。     

内皮素-1与肿瘤血管生成的关系

内皮素-1是一种血管收缩因子，近期研究发现肿瘤细胞和血管内皮细胞可以分泌内皮素-1，促使这些细胞增殖、迁移，抑制其凋亡，并通过促进VEGF分泌，协同作用于肿瘤血管生成。因此，内皮素-1被认为是一种新型血管生长因子，抑制内皮素-1的治疗可以在抗肿瘤血管生成中发挥一定的作用。     

干细胞与肿瘤血管新生

肿瘤血管新生的过程是在一系列正反馈和负反馈因子作用下发生的复杂的链级反应。血管干细胞是产生血和内皮细胞的前体细胞。由于它特有的生物学特性,其在肿瘤血管新生方面的作用受到广泛关注。近年来,大量的研究表明,干细胞参与并促进肿瘤血管新生,从而促进肿瘤的生长和转移。而修饰后的干细胞则可以起到反向的作用。本文就肿瘤血管新生的调控,干细胞的特性及其在肿瘤血管新生过程中的作用和潜在的应用价值作一综述。     

缺氧诱导因子1α和血管内皮生长因子表达及其与肺癌生物学行为的相关性研究

目的探讨缺氧诱导因子-1α(HIF-1α)和血管内皮生长因子(VEGF)表达及其与肺癌生物学行为的相关性。方法采用免疫组织化学方法对68例肺癌组织和50例正常肺组织的HIF-1α及VEGF进行检测。结果肺癌组织中HIF-1α阳性率为69.1%,显著高于正常肺组织(P<0.01);肺癌组织与正常肺组织中VEGF阳性率分别为70.6%和4.0%,差异有统计学意义(P<0.01);HIF-1α和VEGF表达与肺癌组织的临床分期、组织分化及淋巴转移密切相关(P<0.05);肺癌组织HIF-1α和VEGF的表达显著相关(P<0.05)。结论 HIF-1α和VEGF参与了肺癌的发生发展,与肿瘤组织分化、侵袭转移等生物学行为密切相关,可作为反映肺癌预后的重要指标。     

HIF-1α和VEGF在大肠癌中的表达及其相关性研究

[目的]探讨HIF-1α和VEGF在大肠癌浸润、转移过程中的表达及其相互作用机制。[方法]采用免疫组化SP法检测90例大肠癌组织及其相应癌旁组织中HIF-1α和VEGF的表达。[结果]癌旁组织中HIF-1α的表达均为阴性,大肠癌组织中的阳性表达率为73.33%（66/90）,显著高于癌旁组织,差异有统计学意义（P〈0.05）。HIF-1α在大肠癌中阳性表达率与临床分期、浸润深度和淋巴结转移有关（P〈0.05）,与大肠癌分化程度无关（P〉0.05）。癌旁组织中VEGF的表达均为阴性,在大肠癌组织中的阳性表达率为70.00%（63/90）,显著高于癌旁组织,差异有统计学意义（P〈0.05）。VEGF在大肠癌中阳性表达率与临床分期、浸润深度和淋巴结转移有关（P〈0.05）,与大肠癌分化程度无关（P〉0.05）。大肠癌组织中HIF-1α和VEGF蛋白表达具有正相关性（r=0.592,P〈0.05）。[结论]HIF-1α与VEGF的表达率均随着大肠癌浸润深度、淋巴结转移和临床分期的发展而增高,共同参与了大肠癌的发生、发展,可作为大肠癌浸润和转移的重要判定指标。HIF-1α可能通过上调VEGF的蛋白表达,促进大肠癌血管生成而促进大肠癌的转移,抑制HIF-1α的表达可能是治疗大肠癌的新手段。     

缺氧诱导因子-1与卵巢肿瘤

缺氧诱导因子-1(HIF-1)是细胞在缺氧的环境下产生的具有转录活性的核蛋白,其能够使组织对缺氧产生适应性反应,并与包括卵巢肿瘤在内的肿瘤的血管生成、能量代谢、侵袭性转移和预后有关.深入研究HIF-1与卵巢肿瘤之间的关系.探讨其在卵巢肿瘤中的作用和表达机制,将可能为卵巢肿瘤的治疗提供一条新途径.     

Influence on Biological Behavior of Colon Cancer Stem Cells after RNA Interfering CD133

OBJECTIVE To observe the effects of blocking CD133 gene expression and activation on biological characteristic of the colon cancer stem cells.METHODS CD133+ colon cancer stem cells (CCSCs) were separated from EpCAMhigh CD44+ CCSCs through fluorescenceactivated cell sorting (FACS). The proliferation, the capability of spherical cell formation, neoplasia, and the expression of ABCG2 mRNA of CD133+ CCSCs were observed after the CD133+ CCSCs were infected with LV-CD133shRNA. CD133 negative cells were isolated from EpCAMhigh CD44+ CCSCs with FACS, and the CD133 proteins in CD133- cells were detected with Western blot.RESULTS CD133+ CCSCs were isolated from EpCAMhigh CD44+ CCSCs using FACS, and they accounted for 89.2% in the stem cells. In the experimental group, after the CD133+ CCSCs were knocked down by LV-CD133shRNA RNAi, the growth pattern of the cells in the stem cell culture changed into adherent growth from suspended growth, and couldn't generate spherical cells. Results of MTT assay showed that the CD133+ CCSCs infected with LV-CD133shRNA grew slowly, compared to the cells in the control groups. There was a decrease in the cloning efficiency. The infected cells were transplanted into the BALB/c nude mice. During the observation, no neoplasia was found in the CD133+ cells infected with LV-CD133shRNA. The level of ABCG2 mRNA expression was lowered greatly (P < 0.01). CD133- cells were obtained from the EpCAMhigh CD44+ CCSCs using FACS, in which the expression of CD133 protein was positive.CONCLUSION CD133 retains the biological characteristics of the colon cancer stem cells.     

NID1增强人卵巢癌细胞OVCAR-3干细胞特性及分子机制研究

摘 要：［目的］ 探讨NID1对卵巢癌干细胞表型的潜在影响以及NID1与干细胞标志蛋白联合对预后的潜在价值。［方法］ 采用肿瘤干细胞球形成实验、荧光定量RT-PCR和Western blot分别检测稳定过表达外源NID1的OVCAR-3细胞的自我更新能力和卵巢癌干细胞标志物的表达情况。借助生物信息学分析评估NID1在卵巢癌临床样本中与卵巢癌干细胞标志物的相关性及两者联合对预后预测的价值。［结果］ 与空载细胞相比,稳定过表达NID1的OVCAR-3细胞自我更新能力增强（P=0.006）,同时伴随着干细胞标志物CD44（P=0.000）和ABCG2（P=0.000）表达水平升高。经ERK/MAPK通路的抑制剂U0126下调ERK1/2的磷酸化水平后,其CD44和ABCG2的水平下降。此外,基于两套卵巢癌表达谱芯片数据的生物信息学分析表明,NID1的表达水平与CD44（P=0.006;P=0.036）、ABCG2（P=0.000;P=0.000）表达水平均显著正相关,而NID1高表达并且携带干细胞表型的卵巢癌患者总体存活时间最短（P=0.026;P=0.000）。［结论］ NID1可能通过激活ERK/MAPK通路诱导卵巢癌细胞出现干细胞特性,NID1与干细胞标志物联合可能具有预后预测意义。     

Suitability of CD133 as a Marker for Cancer Stem Cells in Melanoma

Objectives: CD133 is considered a cancer stem cell (CSC) marker in various malignancies; however, its role as a biomarker of malignant melanoma remains controversial. The present study was conducted to evaluate the suitability of CD133 surface antigen as a CSC marker in melanoma. Methods: Human melanoma cells were fractionally separated by magnetic cell separation depending on the CD133 phenotype and transplanted into immunodeficient mice to evaluate their tumorigenic capacity. Furthermore, the time until the development of a palpable tumor and the growth rate were measured, and the final tumor volume was assessed after 8 weeks. The immunohistochemical expression of CD133 in the induced neoplasia was then compared using histomorphometry. Results: Notably, neoplasms were induced in all the groups (n = 48), including in the CD133-negative group. Tumors induced by unsorted cells had the largest volume (p = 0.014) but were detected significantly later in this group (p ≤ 0.001). Interestingly, all explanted tumors expressed CD133, with no significant differences among groups. Conclusions: In contrast to the results obtained in prior studies, the suitability of CD133 as a CSC marker could not be demonstrated. The current encouraging progress in targeted therapy for malignant melanoma highlights the need to identify more effective targets.     

BMP4 Administration Induces Differentiation of CD133+ Hepatic Cancer Stem Cells, Blocking Their Contributions to Hepatocellular Carcinoma

CD133(+) cancer stem cells (CSC) contribute to hepatocellular carcinoma (HCC) progression and resistance to therapy. Bone morphogenetic protein BMP4 plays an important role in hepatogenesis and hepatic stem cell differentiation, but little is known about its function in hepatic CSCs. In this study, we showed that high-dose exogenous BMP4 promotes CD133(+) HCC CSC differentiation and inhibits the self-renewal, chemotherapeutic resistance, and tumorigenic capacity of these cells. Interestingly, we found that low-dose exogenous BMP4 upregulated CD133 protein expression in vitro, and endogenous BMP4 was preferentially expressed in CD133(+) HCC CSCs, suggesting that low doses of BMP4 may facilitate CSC maintenance. A reduction in endogenous BMP4 levels decreased CD133 protein expression in vitro. In HCC tissues, expression of the BMP4 signaling target gene SMAD6 was positively correlated with CD133 expression. Activation of the Erk1/2 signaling pathway led to BMP4-mediated reduction in CD133 expression, which was reversed by treatment with MEK inhibitors. Taken together, our findings indicated that BMP4 might be a potent therapeutic agent in HCC that targets CSCs. Cancer Res; 72(16); 4276-85. ?2012 AACR.     

乳腺浸润性导管癌组织中ALDH1+/CD133+干细胞样细胞与血管生成的关系

目的 探讨乳腺浸润性导管癌组织中肿瘤干细胞标志物ALDH1、CD133的表达及其与肿瘤血管生成的关系。方法 应用免疫组织化学双染法检测120例乳腺浸润性导管癌组织中ALDH1⁺/CD133⁺ 干细胞样细胞,单染法检测血管性标记CD34、CD105及VEGF的表达情况。统计ALPH1⁺/CD133⁺干细胞样细胞与临床病理因素;CD34、CD105与VEGF的关系。结果 25.83％（31/120）的病例存在ALDH1⁺/CD133⁺干细胞样细胞,ALDH1⁺/CD133⁺干细胞样细胞与ER、VEGF的表达及MVD均相关（P＜0.05）,但与年龄、肿瘤直径、PR、Her-2、组织学分级及淋巴结转移均无关（P＞0.05）。结论 乳腺浸润性导管癌组织中ALDH1⁺/CD133⁺干细胞样细胞可能通过调节VEGF的表达促进肿瘤新生血管的生成。     

Liposarcoma Cells with Aldefluor and CD133 Activity have a Cancer Stem Cell Potential

ABSTRACT: Aldehyde dehydrogenase (ALDH) has recently been shown to be a marker of cancer stem-like cells (CSCs) across tumour types. The primary goals of this study were to investigate whether ALDH is expressed in liposarcomas, and whether CSCs can be identified in the ALDHhigh subpopulation. We have demonstrated that ALDH is indeed expressed in 10 out of 10 liposarcoma patient samples. Using a liposarcoma xenograft model, we have identified a small population of cells with an inducible stem cell potential, expressing both ALDH and CD133 following culturing in stem cell medium. This potential CSC population, which makes up for 0,1-1,7 % of the cells, displayed increased self-renewing abilities and increased tumourigenicity, giving tumours in vivo from as few as 100 injected cells.     

Targeting the BRAF Signaling Pathway in CD133pos Cancer Stem Cells of Anaplastic Thyroid Carcinoma

Background: Cancer stem cells (CSCs) with a self-renewal ability in tumor cells population, execute a pivotalfunction in tumorigenesis, retrogression, and metastasis of malignant cancers such as anaplastic thyroid carcinoma(ATC). Materials and Methods: In this study, we isolated CSCs subpopulation with CD133 surface marker fromthree ATC cell lines by magnetic cell sorting assay. After confirming the segregation by the flow cytometry method,BRAF and sodium-iodide symporter (NIS) genes were investigated in them before and after incubation with BRAFinhibitor. Also, we evaluated the NIS protein expression and localization. Results: Established upon q-RT PCR data,when compared to human normal thyrocytes, the BRAFV600E gene was over-expressed in CD133pos cells (>1705.99 ±55.55 fold, Mean ± SEM, n=3, P- value<0.05), whilst the expression of NIS gene was very restricted (< 0.0008 ± 5.43fold, Mean ± SEM, n=3, P- value<0.05) in them. Also, our results showed that BRAF inhibition affected NIS proteinexpression and localization. Conclusions: Current study showed that the differentiate genes/proteins expression canbe induced in the CSCs via focus on signal transduction pathways and targeting their molecules, that are involved inexpression of these genes/proteins. Therefore, attention to targeting CSCs along with routine thyroid cancer therapy,can help to ATC treatment.     

CD44、CD133耦连miR-425装载纳米脂质体对结直肠癌干细胞PDL-1表达的下调作用

目的 探究CD44、CD133耦连miR-425装载纳米脂质体对结直肠癌干细胞PDL-1表达的下调作用.方法选择结直肠癌细胞系HCT116分选出CD44、CD133/1+细胞,进行miR-425装载纳米脂质体转染,分为对照组(转染con-trol载体)、实验组(转染miR-425慢病毒载体),空白组以纳米脂质体代替,测定干细胞生长状态,检测PDL-1表达水平.结果分选的CD 133/1+细胞呈圆形,12 h后开始贴壁生长,状态良好.qRT-PCR检测显示实验组的miR-425的相对表达水平为(182.44±22.19),对照组与空白组分别为(1.84±1.11)和(1.67±0.98),实验组明显高于对照组与空白组(P<0.05).MTT检测结果表明,与空白组(0.78±0.22)、对照组(0.81±0.17)比较,实验组(0.24±0.15)的HCT116细胞生长速度明显减慢(P<0.05).空白组与对照组的细胞增殖率分别为(0.48±0.14)与(0.51±0.18),都明显高于实验组的(0.30±0.11).Western blot检测分析显示实验组、对照组、空白组的PDL-1相对表达量分别为(0.33±0.13)、(0.76±0.21)和(0.82±0.15),实验组明显低于对照组与空白组(P<0.05).结论CD44、CD133耦连miR-425装载纳米脂质体能抑制结直肠癌干细胞PDL-1的表达,从而显著抑制干细胞的生长、增殖能力.     

CD133作为胶质瘤干细胞标志物应用的局限性

胶质瘤是神经外科最常见的恶性肿瘤,临床尚缺乏有效的治疗手段。肿瘤干细胞被认为是胶质瘤发生、发展的源头而受到越来越多的关注。干细胞标志物CD133广泛表达于人体多种肿瘤中,同时,它也是研究得最多且应用最广的胶质瘤干细胞表面标志物。然而,利用CD133进行胶质瘤干细胞的鉴定和筛选正逐渐受到质疑。本文将对近年来人们发现的CD133作为胶质瘤干细胞标志物应用的局限性进行综述。     

Expression of CD133 Cancer Stem Cell Marker in IDH-Mutant and IDH-wildtype (Isocitrate Dehydrogenase) Astrocytoma

Objective: This study evaluated the differences between IDH1-R132H and CD133 expression in different categories of  astrocytoma. Material and methods: This study used a cross-sectional design.  Sixty-seven paraffin embedded block of Diffuse Astrocytoma (DA), Anaplastic Astrocytoma (AA) and Glioblastoma (GB) were assessed using using the monoclonal antibody IDH1-R132H and Rabbit polyclonal antibody CD133. Results: It was found that there was a significant relationship between the expression of IDH1-R132H and CD133 in DA, AA and GB (p<0.001). Astrocytoma with IDH-mutant molecular status will express more markers of cancer stem cell CD133 than IDH-wildtype. Conclusion: The IDH1-R132H and CD133 can provide predictive value on treatment success, disease prognosis, recurrence and can be considered as target combination therapy with chemotherapy.