CD105对绒毛膜癌裸鼠移植瘤生长及耐药性的影响

目的:探讨绒毛膜癌裸鼠移植瘤模型中的成瘤情况,以及CD105及相关蛋白在移植瘤中表达水平与化疗耐药的相关性.方法:将90只健康BALB/c裸鼠随机分为3组:JEG-3细胞(野生型)、JEG-3/CD105 OE(过表达型)与JEG-3/shCD105(敲减表达型),每组30只,建立CD105表达相关的绒癌裸鼠移植瘤模型...     

宫颈癌组织CD105表达与局部血管生成、癌细胞增殖及侵袭的关系

CD105(endoglin)相对分子质量为180000，是一种存在于细胞表面的同源二聚体跨膜糖蛋白，能与多种转化生长因子(TGF)超家族成员如TGF-β结合，通过调节TGF-β与其受体结合，参与信号传导及血管生成。CD105基因定位于人9号染色体，是Ⅰ型遗传性出血性毛细血管扩张症的相关基因，可能与血管生成启动有关。本研究采用免疫组织化     

早发型子痫前期可溶性CD105的表达及变化意义

子痫前期(preeclampsia)是孕产期特有的疾病,典型症状为逐渐加重的高血压、水肿、蛋白尿等,是严重危害母儿健康的产科并发症,在我国发病率9.4%～10.4%,易导致胎儿宫内缺氧,是围产儿死亡的主要原因,发病机制至今不明。越来越多的研究发现,血管生成因子与子痫前期的发生发展有密切的关系。本文就可溶性CD105淋巴细胞抗原(solubleendoglin,sEng)在早发型子痫前期发病机制中的作用及其对子痫前期的预测作用进行综述。研究表明,在子痫前期患者发病的前几周,血清中的血管生成因子水平较正常妊娠的孕妇已有显著变化。有学者研究证实CD105浓度在晚发型和早发型子痫前期的患者中均显著升高,且在早发型患者中尤其显著。血清sCD105水平的升高可能是早发型子痫前期发病的重要机理。     

细胞自噬在甲氨蝶呤耐药绒癌中的保护性作用

目的：通过检测耐药绒癌细胞内自噬的变化,以及评估抑制自噬或敲除自噬相关基因后细胞凋亡情况,探讨细胞自噬在耐药绒癌中的作用。方法：以人绒毛膜上皮癌JEG-3细胞及本课题组前期构建的甲氨蝶呤（MTX）耐药绒癌JEG-3／MTXR细胞为研究对象。采用透射电镜、免疫荧光法观察MTX处理后细胞内自噬的变化;Westernblot法检测自噬相关蛋白LC3、Bec]in-1的表达变化。RNA干扰法沉默自噬相关基因LC3后,采用Westernblot法和Annexin-V／PI双染法流式细胞仪检测MTX作用后JEG-3／MTXR耐药绒癌细胞凋亡的变化。结果：透射电镜和荧光显微镜下均可观察到MTX处理后JFG-3、JEG-3／MTXR细胞内自噬小体的产生;Icm浓度MTX处理后,两种细胞内自噬相关蛋白LC3、Beclin-1表达水平上调。相同浓度MTX处理后,JEG-3细胞内LC3、Beclin-1表达下调。沉默自噬相关基因LC3后,沉默LC3联合MTX组的JEG-3／MTXR和JEG-3细胞内凋亡相关蛋白caspase-9表达上调,凋亡率显著增多。结论：MTX可诱导耐药绒癌产生保护性自噬。     

宫颈鳞癌中膜突蛋白及CD105的表达及其临床意义

目的:研究膜突蛋白(Moesin)及CD105在各级宫颈病变中的表达水平,探讨其与宫颈鳞癌的发展及浸润的关系。方法:采用免疫组化SP法检测31例正常宫颈组织(NCE)、48例宫颈上皮内瘤样变(CIN)及66例宫颈鳞癌组织(SCC)中Moesin及CD105的表达。结果:从NCE到CIN到SCC组,Moesin的表达水平显著升高。在CIN组Moesin的表达随病变程度加重而升高;SCC组中与FIGO分期、浸润深度、盆腔淋巴转移有关,与组织分化程度及年龄无关。从NCE到CIN到SCC组,CD105表达的微血管密度(MVD)值显著升高,在SCC组中与淋巴转移及浸润深度有关,且与Moesin蛋白具有较高关联性。结论:Moesin蛋白及CD105可能在宫颈鳞癌的发展及转移过程中起重要作用,二者结合可能成为判断宫颈病变生物学行为的重要指标。     

内质网应激介导的凋亡耐受在甲氨蝶呤耐药绒癌发生机制中的作用

目的:初步探讨内质网应激介导的凋亡耐受在甲氨蝶呤耐药绒癌发生机制中的作用。方法:以人绒毛膜上皮癌JEG-3细胞及本课题组前期构建的甲氨蝶呤耐药绒癌细胞(JEG-3/MTXR)为研究对象。采用WST法评估甲氨蝶呤处理后JEG-3/MTXR和JEG-3亲本细胞的细胞活力,Western blot法测定两种细胞中caspase-9激活、GRP78和GADD153蛋白的表达。RNA干扰法沉默JEG/MTXR细胞中GADD153基因,Western blot法测定caspase-9蛋白激活的情况。结果:不同浓度梯度的甲氨蝶呤诱导后,JEG-3亲本细胞系的细胞生存率显著降低。相同浓度甲氨蝶呤作用后,JEG-3/MTXR细胞内切割caspase-9无增加,JEG-3亲本细胞内caspase-9激活并切割。JEG-3/MTXR和JEG-3细胞中GRP78表达水平呈时间依赖性上调;JEG-3/MTXR细胞中GADD153蛋白水平无显著上调,而JEG-3亲本细胞内出现显著上调。沉默GADD153基因可抑制MTX引起的切割caspase-9蛋白水平上调。结论:内质网应激介导的凋亡耐受可能参与甲氨蝶呤耐药绒癌发生机制。     

6种常见耐药标志物在氟尿苷耐药绒癌细胞系构建过程中的动态表达

目的:通过检测对氟尿苷(FUDR)耐药的绒癌细胞系构建过程中肺耐药相关蛋白(LRP)、二氢叶酸还原酶(DHFR)、survivin、谷胱甘肽 S-转移酶-π(GST-π)、多药耐药基因(MDR1)和多药耐药相关蛋白基因(MRP)6种常见耐药标志物的动态表达,探讨其耐药机制并进行耐药标志物的初步筛选.方法:用化学发光法和荧光定量PCR技术,分别检测不同浓度FUDR诱导的JEG-3细胞中β-人绒毛膜促性腺激素(β-HCG)的分泌量和上述6种常见耐药标志物在耐药细胞构建过程中的动态表达.结果:不同基因在药物诱导过程中的变化趋势不尽相同.只有LRP基因的表达随着诱导药物浓度和肿瘤细胞耐药性的增加而逐渐上调.结论:耐药的产生是分阶段的,不同阶段有不同的因素参与.就目前的研究结果而言,只有LRP的表达变化与FUDR诱导浓度和耐药性有明显正相关,提示其可以作为绒癌对FUDR耐药的候选预测指标.     

CD105在妇科恶性肿瘤中的应用及研究进展

肿瘤血管的生成在肿瘤的生长和转移中至关重要,其可以满足肿瘤生长的代谢需要并为远处转移提供血管通路,因此被认为与肿瘤的预后密切相关。CD105又名Endoglin,是转化生长因子β（TGF-β）受体超家族的成员之一,在标记肿瘤新生血管中具有较好的特异性。近年来,CD105越来越多地应用于妇科恶性肿瘤的诊断及预后判断,其在血管内皮细胞以外的肿瘤实质及间质细胞中的作用及机制正逐步被揭示,全面了解CD105的生物学特性将为探究妇科恶性肿瘤的发生发展机制、预防、诊断及治疗提供新的思路和治疗靶点。     

人绒毛膜促性腺激素和胸苷合成酶在氟尿嘧啶核苷连续诱导耐药绒癌细胞系构建过程中的动态表达

目的:通过检测氟尿嘧啶脱氧核苷(fluorodeoxyuridine,FUDR)连续诱导的耐药绒癌细胞构建过程中β-人绒毛膜促性腺激素(β-human chorionic gonadotropin,β-HCG)和胸苷合成酶(thymidylate synthase,TS)的动态表达,检验β-HCG分泌水平和TS基因的表达量是否适用于筛查绒癌细胞对该药耐药的指标。方法:采用连续诱导法,利用FUDR诱导人绒癌细胞系JeG-3,建立耐药细胞株JeG-3/FUDRC2;用化学发光法检测培养液上清中激素分泌水平,用荧光定量PCR检测TS mRNA在诱导过程中的动态表达。结果:(1)历时12月成功建立了绒癌耐药细胞系JeG-3/FUDRC2,其耐药指数为12.46±2.22;(2)随着药物浓度和肿瘤细胞耐药性的逐渐增加,绒癌细胞的β-HCG分泌最初表现为上调,诱导药物达到一定浓度时其分泌达到高峰,而后骤然下降,并有随药物浓度和耐药性的继续增加而逐渐下降的趋势;(3)低浓度药物诱导后,TS基因表达明显上调,但随着药物浓度和耐药性增加,其表达不升反降,甚至低于亲本细胞TS基因的表达量。结论:成功建立了连续诱导耐药的绒癌细胞株JeG-3/FUDRC2。绒癌细胞β-HCG分泌和TSmRNA表达与FUDR作用浓度以及耐药性无明显相关性。就目前的研究结果而言,β-HCG分泌和TS mRNA的表达水平不适宜用作绒癌细胞是否对FUDR耐药的指标。     

CD4、CD8、FoxP3在子宫颈癌新辅助化疗前后表达的变化及临床价值

目的:探讨CD4、CD8、FoxP3在子宫颈癌新辅助化疗(NACT)前后表达的变化及临床价值。方法:选择上海交通大学医学院附属仁济医院妇产科2013年10月至2017年9月收治的应用NACT的局部晚期(ⅠB2～ⅡB)子宫颈癌患者NACT前后免疫组化检测CD4+、CD8+及FoxP3表达的变化,并通过计算机软件进行定量分析其与化疗疗效、无病生存期(PFS)、总体生存期(OS)之间的关系。结果:局部晚期子宫颈癌NACT前后CD4、CD8、FoxP3在组织中表达的变化与患者年龄、病灶大小无相关性(P0. 05),但是CD4、CD8与子宫颈癌FIGO分期呈正相关(P 0. 05),与组织病理学分级呈负相关(P 0. 05)。NACT后CD4、CD8、FoxP3的平均单位面积阳性值较化疗前显著升高(P=0. 016,P=0. 009,P=0. 005);局部晚期子宫颈癌患者NACT疗效与CD4、CD8和FoxP3表达的变化无明显相关性(P0. 05),但NACT后CD8表达的增加与患者PFS呈正相关(r=0. 309,P=0. 041),而与CD4(P=0. 581)、FoxP3(P=0. 134)无明显关系。患者的OS与CD4、CD8和FoxP3化疗前后表达的变化无明显相关性(P 0. 05)。结论:局部晚期子宫颈癌NACT后,CD4、CD8、FoxP3表达较NACT前升高,CD4、CD8表达的变化与肿瘤分期、病理分级相关,而且CD8化疗前后的变化可以作为独立的预后指标。     

The prognostic value of endoglin (CD105) expression in ovarian carcinoma

Intratumoral angiogenesis has become an important issue after the identification of antiangiogenic therapeutics. The purpose of this study was to investigate the prognostic value of CD105 in patients with ovarian cancer and also to compare with CD31. Fifty-eight patients were included to this study. All the paraffin blocks were reviewed, and angiogenesis was determined by immunohistochemical staining, using anti-CD105 and anti-CD31 monoclonal antibodies. The mean microvessel density (MVD) with CD105 and CD31 were 28.78 +/- 22.20 and 28.69 +/- 18.57, respectively (P = 0.97). With respect to prognostic factors, CD31 was only significant for suboptimal cytoreduction (P = 0.02), and CD105 was significant for both advanced stage and suboptimal cytoreduction (P = 0.02 and P = 0.05, respectively). For survival analysis, patients were divided into three groups by quartiles for each marker (group 1, <25%; group 2, 25-75%; and group 3, >75%). By CD31, only significant difference was noted between group 1 and group 2 (P = 0.03). In analysis with CD105, the survival rate of patients with group 3 was significantly worse than group 1 and group 2 (P = 0.01 for both). In multivariate analysis, cytoreduction and MVD determined by CD105 remained significant. In this study, endoglin was found to be an independent predictor of poor survival. Therefore, it could be used for antiangiogenic therapies.     

CD105 expression is an independent predictor of survival in patients with endometrial cancer

OBJECTIVE: The purpose of this study was to detect the prognostic value of CD105 (endoglin) and also to compare with CD34 and vascular endothelial growth factor (VEGF) in patients with endometrial adenocancer. METHODS: Ninety patients with endometrial carcinoma, who were treated at Gazi University Hospital, were included. Staging was performed according to the FIGO recommendations. Angiogenesis was estimated by using CD105 and CD34 and tested for possible significant relation with age, stage, histologic type, grade, depth of myometrial invasion, lymphovascular space invasion, lymph node metastasis, and overall survival (OS). In addition, VEGF staining intensity and distribution were analyzed with respect to all these variables. RESULTS: The mean age at the time of diagnosis was 57.7 years (range, 28-81 years). The mean microvessel density (MVD) for CD105 was 32.87+/-19.99, and it was 55.46+/-31.25 for CD34 (P<0.001). A significant correlation was noted between these two markers (r=0.257, P=0.02). The mean VEGF score was 4.13+/-1.73, and it was significantly correlated with MV counts determined by CD105 (r=0.291, P=0.006). It was not significantly related with CD34 (r=0.031, P=0.78). With respect to clinicopathological variables, none of the comparisons was found to be significant. The mean follow-up period was 60.5 months. To analyze the prognostic value of MVD, the patients were divided into three groups with respect to quartiles (or=75%). With CD105 staining, the 5-year OS rates for patients with the highest MVD count (>or=75%) were significantly poorer than the remaining two groups (P=0.01 for both). None of the comparisons for CD34 was significant. Survival analysis for VEGF was performed by grouping patients using staining characteristics. No significant difference was noted for OS. Multivariate analysis showed that MVD determined by CD105 correlated significantly and independently with OS (P=0.02). None of the remaining variables was significant in multivariate analysis. CONCLUSION: The current study showed that CD105 is an independent predictor of survival in patients with endometrial cancer. We recommend the use of this highly specific and prognosis-related antigen in further investigations.     

新生儿感染时CD分子变化及临床意义

目的 探讨不同胎龄新生儿感染时T、B、NK细胞及中性粒细胞表面CD分子变化及临床意义。方法 2004年2~6月复旦大学儿科医院用流式细胞仪检测34例早产儿及33例足月儿CD表达。结果(1)足月感染组CD3［(76.89±2.52)%］高于足月非感染组［(64.40±5.69)%］，早产非感染组［(80.93±9.13)%］高于足月非感染组；早产非感染组CD4［(61.20±2.21)%］高于足月非感染组［(47.60±4.27)%］，早产感染组［(53.63±3.23)%］低于早产非感染组；CD8各组间无统计学差异性。(2)CD19各组间无统计学差异性。(3)足月感染组CD56CD16［(5.88±0.62)%］低于非感染组［(13.00±5.31)%］，足月非感染组高于早产非感染组［(6.13±1.25)%］。(4)足月感染组CD64［(5056.92±1255.58)分子/单位］高于足月非感染组［(2112.60±1157.21)分子/单位］，早产感染组［(4619.67±1395.99)分子/单位］高于早产非感染组［(2407.45±1247.16)分子/单位］。(5)败血症、肺炎及其它感染组CD64高于非感染组。以CD64大于3000分子/单位为阳性，CD64诊断感染灵敏度为79.6%，特异度为75.1%；以CD64大于2000分子/单位为阳性，CD64诊断感染灵敏度为88.9%，特异度为60%；CRP诊断感染灵敏度为65.3%，特异度为86%。结论早产儿细胞表面CD3、CD4、CD16CD56变化不同于足月儿，可能与早产儿免疫功能低下有关；CD64可能会成为一种新型感染诊断指标。 Abstract ObjectiveTo evaluate the change and the clinical significance of the cell CD on the surface of T,B,NK and Neutrophils cells.MethodsFrom Feb. to June 2004,50 preterms and 49 terms with infection or non infection were sturied.The level of peripherical blood CD3,CD4,CD8,CD19,CD16CD56 and CD64 were measured by flow cytometry.Results① The level of CD3 in terms(76.89±2.52)% with infection was higher than that in terms without infection(64.40±5.69)%.The level of CD3 in preterms without infection(80.93±9.13)%was significantly higher than that in terms without infection.The level of CD4 in preterms without infection(61.20±2.21)% was significantly higher than that in terms without infection(47.60±4.27)%.The level CD4 in preterms with infection (53.63±3.23)% was significantly lower than that in preterms without infection ；The level of CD8 was not different in all groups.②The level of CD19 has no difference in all groups.③The level of CD56CD16 in terms with infection(5.88±0.62)% was significantly lower than that in terms without infection(13.00±5.31)%，the level of CD56CD16in terms without infection was significantly higher than that in preterms without infection(6.13±1.25)%.④ The level of CD64 in terms with infecton (5056.92±1255.58)Molecule/Unit was higher than that in terms without infection (2112.60±1157.21)M/U.The level of CD64(4619.67±1395.99)M/U in preterms with infection was significantly higher than that in the preterms without infection(2407.45±1247.16)M/U.⑤The level of CD64 in patients with sepsis,pneumonia and other infections were higher than that in those patients without infection.If the positive standard of CD64 was over 3000 M/U,then the sensitivity was 79.6% and the specificity was 75.1%.If the positive standard of CD64 was over 2000 M/U,then the sensitivity was 88.9% and the specificity was 60%.The sensitivity of CRP was 65.3% and the specificity was 86%.Conclusion The level of CD3、CD4、CD16CD56 in preterms with or without infection differs from those in terms,which is probably due to the low immunity function of preterms.CD64 may be a kind of new diagnosis guideline. Key wordsTerms;Preterms;CD molecule;CD64     

乙酰肝素酶与绒毛膜癌侵袭能力的关系

目的 通过研究乙酰肝素酶（Hpa）在绒毛膜癌高、低侵袭力细胞系及正常早孕绒毛组织中的表达情况,探讨Hpa与绒毛膜癌侵袭能力的关系。方法 通过Matrigel体外侵袭实验检测不同侵袭力绒毛膜癌细胞系JEG-3和JAR的体外侵袭能力;应用免疫细胞化学方法及蛋白印迹法（westem blot）检测Hpa蛋白在不同绒毛膜癌细胞系及正常早孕绒毛组织中的表达,并进行相关性分析。结果 （1）JEG-3细胞的穿膜细胞数[（191±17）个]较JAR细胞的穿膜细胞数[（106±13）个]明显增多,两者比较,差异有统计学意义（P〈0．05）。（2）Hpa蛋白在绒毛膜癌细胞JEG-3、JAR中均有表达,且主要定位于细胞质。（3）在正常早孕绒毛组织、JEG-3细胞和JAR细胞中均可检测到Hpa蛋白,JEG-3细胞中Hpa蛋白的表达量（1．560±0．180）明显高于JAR细胞Hpa蛋白的表达量（0．610±0．170）,绒毛膜癌细胞系中Hpa蛋白的表达量又显著高于正常早孕绒毛组织Hpa蛋白的表达量（0．190±0．008）,两两比较,差异均有统计学意义（P〈0．05）。（4）相关性分析显示,Hpa蛋白的表达量与绒毛膜癌细胞体外侵袭能力呈正相关关系（r=0．89,P〈0．05）。结论 Hpa蛋白在绒毛膜癌细胞中表达较正常绒毛组织高;Hpa蛋白在滋养细胞中的表达随滋养细胞侵袭能力的增加其表达上调;滋养细胞产生过量的Hpa对基底膜蛋白的水解作用在绒毛膜癌的侵袭、转移中发挥重要作用。     

ABCG2与CD133在宫颈癌的表达及意义

目的:探讨ABCG2及CD133蛋白在宫颈癌的表达及临床意义。方法:用免疫组化SP法检测92例宫颈癌、40例宫颈上皮内瘤变(CIN)和30例正常宫颈组织中AB-CG2及CD133蛋白的表达。结果:ABCG2在宫颈癌、CIN和正常宫颈组织的阳性表达率分别为69.6%、42.5%、23.3%,CD133为57.6%、37.5%、23.3%,两者在宫颈癌的表达均高于CIN及正常组织,差异均有统计学意义(P0.05);两者表达与宫颈癌患者年龄、肿瘤大小、临床分期无相关性(P0.05),而在不同分化组间表达的差异有统计学意义(P0.05)。结论:ABCG2和CD133蛋白异常表达与宫颈癌的发生、发展有关,二者联合检测可作为评估宫颈癌恶性程度、指导治疗的重要参考指标。