RCAS1在宫颈癌组织中的表达及其与HPV16感染的关系

背景与目的:表达在SiSo细胞上的受体结合肿瘤抗原(receptor-binding cancer antigen expressed on SiSo cells,RCAS1)在多种肿瘤组织中呈高表达,并与肿瘤逃避免疫监视有关.本研究检测RCAS1蛋白在宫颈癌组织中的表达及其与HPV16感染的相关性,并探讨其临床意义.方法:采用免疫组化SP(streptavidin-peroxidase)法,分别检测71例宫颈癌、76例宫颈上皮内瘤样病变(CIN)及20例正常宫颈上皮组织中RCAS1蛋白与HPV16 E7蛋白的表达,并分析两者的表达与临床病理因素的关系.结果:宫颈癌组织中,RCAS1蛋白主要表达于癌细胞膜和/或细胞浆,HPV16 E7蛋白主要表达于癌细胞核.正常宫颈上皮组织不表达RCAS1蛋白,CIN与宫颈癌组织中RCAS1蛋白的表达率分别为39.47%和77.46%,HPV16 E7蛋白的表达率分别为0.05%、28.94%和61.97%,提示随着宫颈病变恶性程度的进展,RCAS1与HPV16 E7表达均逐渐增强(P＜0.05).低分化宫颈癌组中RCAS1表达显著高于高、中分化宫颈癌组(P=0.002),但与患者年龄、临床分期及组织学分型无关(P＞0.05);HPV16 E7在鳞癌组中的表达显著高于腺癌组(P=0.000),但与患者年龄、临床分期、组织学分级无关(P＞0.05).RCAS1的表达与HPV16感染在宫颈癌中的表达呈正相关(r=0.780,P=0.000).结论:RCAS1基因在宫颈癌组织中表达增强,RCAS1表达强度与宫颈癌恶性程度相关;RCAS1阳性的宫颈癌组织中存在HPV16感染.     

肿瘤相关抗原RCAS1与恶性肿瘤的关系

引言RCAS1(receptor-binding cancer antigen ex-pressed on SiSo cells)是由Nakashi ma等[1]于1999年发现的一种肿瘤相关抗原,在多种肿瘤组织都有表达,可被22-1-1单克隆抗体识别,可能参与肿瘤的免疫逃避机制,与肿瘤的发生发展及预后有关。22-1-1抗体是用人子宫腺癌细胞系SiSo细胞免疫的小鼠骨髓瘤细胞和脾细胞通过细胞融合方法获得的[2]。1RCAS1概述1.1RCAS1基因定位和蛋白结构研究认为其基因定位在染色体8q23上,Na-kashi ma等[1]分离了编码它的cDNA,此cDNA包括5′端242个核苷酸的非翻译区,639个核苷酸的编码区和3′端179个核苷酸的非翻译区。RCAS1蛋白是一种分子量为40kD的II型跨膜蛋白,由213个氨基酸组成,含有一个N端的跨膜片段,一个C端部分的卷曲螺旋结构。它可通过卷曲螺旋结构形成寡聚体,而且存在可溶性形式。1.2RCAS1在正常组织的分布和表达RNA印迹分析显示RCAS1基因在卵巢、睾丸、前列腺、胸腺、肌肉及心脏有表达,但免疫组化方法未在任何上述器官中发现RCAS1蛋白表达。在小肠、结...     

RCAS1与FAS蛋白在宫颈癌组织中的表达及意义

目的探讨RCAS1与FAS蛋白在宫颈癌组织中的表达、相互关系及意义。方法采用免疫组化学S-P法检测48例宫颈癌、20例宫颈上皮内瘤变（CIN）和10例宫颈良性病变上皮组织中RCAS1及FAS的表达情况。结果宫颈癌组织中RCAS1表达水平明显高于CIN及宫颈良性病变上皮组织（P〈0．05），而FAS蛋白表达水平明显低于CIN及宫颈良性病变上皮组织（P〈0．05）：RCAS1的阳性表达率与宫颈癌分化程度、淋巴结转移有关（P〈0．05），与患者年龄、组织类型及临床分期无关（P〉0．05）：FAS的阳性表达率与宫颈癌分化程度有关（P〈0．05），与患者年龄、组织类型、临床分期及淋巴结转移无关（P〉0．05）；宫颈癌组织中RCAS1与FAS蛋白表达无明显相关性（r＝-0．652，P〉0．05）。结论在宫颈癌的发生发展过程中RCAS1与FAS蛋白可能起重要作用，但两者的作用机制并无相关性。     

非小细胞肺癌患者血清RCAS1水平检测临床意义分析

目的:探讨非小细胞肺癌(NSCLC)患者血清SiSo细胞表达的受体结合肿瘤抗原(RCAS1)水平及其临床意义。方法:采用酶联免疫吸附法(ELISA)定量检测102例NSCLC患者和40例良性肺部疾病患者血清RCAS1水平,并以35名正常人血清作为对照,比较不同分期、有无转移及术后1年复发与否的患者之间是否存在差异。结果:NSCLC组血清RCAS1水平〔(48.6±18.5)U/mL〕显著高于健康对照组〔(13.5±11.8)μg/mL;t=7.56,P=0.000〕和良性肺部疾病组〔(18.3±14.8)μg/mL;t=3.73,P=0.001〕。Ⅲ/Ⅳ期患者血清RCAS1水平〔(60.8±16.3)U/mL〕显著高于Ⅰ/Ⅱ期患者〔(34.6±15.3)U/mL;t=3.56,P=0.000〕。有转移患者和术后1年内复发患者血清RCAS1水平〔(58.3±16.9)、(59.6±14.9)U/mL〕分别高于无转移患者和无复发患者〔(35.6±13.2)、(33.8±13.6)U/mL〕,P值分别为0.002、0.001。结论:NSCLC患者血清RCAS1表达水平有可能作为肿瘤转移和术后复发的预测指标。     

肺癌患者血清中RCAS1水平及其临床意义

目的探讨肺癌患者血清中SiSo细胞表达的受体结合肿瘤抗原(RCAS1)水平并进行分析,为临床诊治提供依据。方法采用酶联免疫吸附试验(ELISA)分别检测90例肺癌患者、60例良性肺部疾病患者和102名健康体检者血清RCAS1水平,并进行比较分析。结果肺癌组血清RCAS1水平与良性肺疾病组和健康对照组比较差异有统计学意义(P<0.05);随病程进展肺癌患者RCAS1的阳性率逐步升高,肺癌Ⅲ、Ⅳ期患者RCAS1阳性率(82.8%、95.8%)明显高于Ⅰ、Ⅱ期患者(42.9%、56.5%)(P值均<0.05);有淋巴结转移的患者RCAS1阳性率(80.6%)明显高于未转移患者(44.4%)(P<0.05)。RCAS1诊断肺癌的敏感度为73.3%,特异性为96.9%,准确性为88.5%。结论 RCAS1对肺癌的辅助诊断有一定的临床价值,可以作为一项新的肺癌标志物应用。     

康莱特联合奥沙利铂胸腔灌注治疗恶性胸腔积液及对RCAS1、VEGF表 达的影响

目的:探讨康莱特联合奥沙利铂胸腔灌注治疗恶性胸腔积液的临床价值以及灌注治疗对恶性胸腔积液中SiSo细胞上的受体结合肿瘤抗原(RCAS1)、血管内皮生长因子(VEGF)表达的影响。方法:选取110例非小细胞肺癌并胸腔积液患者,分为康莱特组35例,奥沙利铂组35例及联合用药组40例,分别收集胸腔注药前及每次注药48h后的胸水标本,采用酶联免疫吸附测定（ELISA）方法检测胸水标本中的RCAS1、VEGF水平。结果:三组疗效比较,联合组优于康莱特组（P<0.01）。生活质量改善方面,联合组优于康莱特组与奥沙利铂组（P<0.05）。三组治疗后的毒副反应比较,康莱特组低于联合组及奥沙利铂组（P<0.05）。三组治疗前、第1周期治疗后、第2周期治疗后胸水RCAS1及VEGF表达水平均有逐渐下降趋势（P<0.05）,联合用药组较其它两组下降更明显（P<0.001）。结论:康莱特联合奥沙利铂胸腔灌注对控制恶性胸腔积液具有一定的临床价值,检测胸水RCAS1、VEGF表达水平可评估康莱特联合奥沙利铂灌注治疗效果。     

氯化锂对白血病THP-1细胞生长的抑制作用及其对Wnt通路的影响

摘 要：目的 研究氯化锂（LiCl）在体外对白血病细胞THP-1增殖、凋亡、周期以及Wnt通路的影响。 方法 氯化锂不同浓度不同时间作用THP-1细胞后,用MTT法检测细胞增殖,流式细胞术检测细胞凋亡和周期分布,Western blot检测细胞内Wnt通路的变化。 结果 氯化锂能抑制白血病THP-1细胞的增殖,诱导细胞凋亡,使细胞周期阻滞于G2/M期,且均呈浓度和时间依赖性。氯化锂不同浓度不同时间作用THP-1细胞后,t-GSK3β、cyclinD1表达不变,p-GSK3β、β-catenin表达升高,短时间低浓度作用后c-myc表达升高,但随作用时间延长和浓度增加,c-myc表达降低。 结论 氯化锂对THP-1细胞有显著的增殖抑制、凋亡诱导、细胞G2/M期阻滞作用,并激活Wnt通路,影响Wnt通路下游蛋白c-myc、cyclinD1的表达。     

受体结合型癌抗原在胃肠道肿瘤免疫逃逸中的作用

应用免疫组化方法检测受体结合型癌抗原(RCAS1)在胃癌和直肠癌中的表达,同时用原位末端标记法(TUNEL)分析胃癌组织和直肠癌组织中淋巴细胞凋亡情况,以探讨RCAS1与胃肠腺癌发生发     

E-钙黏蛋白在结肠癌侵袭转移机制中的研究进展

结肠癌的侵袭行为与侵袭抑制因子——E-钙黏蛋白表达减少、缺失密切相关。E钙黏蛋白通过与肿瘤相关抗原RCAS1、抗黏附蛋白dysadherin、β-酪蛋白肽等相互作用,与结肠癌侵袭转移机制密切相关,其间涉及免疫逃避机制、肿瘤转移的细胞分子机制、肿瘤信号传导通路、细菌与肿瘤关系等近年研究热点。     

吐温80对BGC—823细胞抑制作用及凋亡相关因子的影响

目的 观察吐温80对人胃癌细胞BGC－823的抑制作用及凋亡相关因子的影响。方法 采用MTT法,测定不同浓度（0．15％、0．1％、0．05％）的吐温80及其不同作用时间（2小时、18小时、34小时）条件下,对BGC－823细胞的抑制作用;运用免疫组化ABC法染色,观察0．1％吐温80％作用（2小时、18小时、34小时）后BGC－823细胞Hsp70、bcl-2、bax的表达。结果 （1）吐温80的抑制效应随作用浓度及时间增加而明显增高（P＜0．01）;（2）吐温80作用不同时间对细胞Hsp70表达及分布影响不明显,而bcl-2、bax随作用时间延长有表达增加趋势,作用34小时,bcl-2主要分布于胞浆,bax主要分布于胞核。结论 吐温80对肿瘤细胞具有较强抑制作用,吐温80对bcl-2、bax表达及分布影响可能与其抑制作用有关。     

11种化疗药物对卵巢癌细胞的抑制作用及其对有核细胞毒性的研究

目的通过体外实验,观察11种化学治疗药物对人卵巢癌细胞的杀伤作用,及其对外周血有核细胞(白细胞、淋巴细胞和粒细胞)的毒性效应,并探讨这2种作用可能存在的相关性。方法采用经典的MTT比色法分别检测34例卵巢癌患者新鲜肿瘤组织标本的癌细胞对11种化疗药物的敏感性和药物对外周血有核细胞的毒性。结果各药物对癌细胞抑制率的差异有显著性;其中,TAX和VM-262种化疗药物的抑制率分别达到50.97%和41.38%,可认为属于卵巢癌细胞的敏感药物;其余9种药物对卵巢癌细胞的抑制率均较低。各药物对癌细胞的抑制率与其对有核细胞中的粒细胞的毒性作用有明显的相关性。结论TAX和VM-26,属于卵巢癌细胞敏感的化疗药物;检测的11种化疗药物,其对卵巢癌的疗效与其对粒细胞的毒性显著相关;在缺乏进行肿瘤细胞药敏试验条件的情况下,卵巢癌患者外周血粒细胞毒性试验,是1种可行的替代检测措施。     

PDGF upregulates CLEC-2 to induce T regulatory cells

The effect of platelet derived growth factor (PDGF) on immune cells is not elucidated. Here, we demonstrate PDGF inhibited the maturation of human DCs and induced IL-10 secretion. Culture of PDGF-DCs with T cells induced the polarization of T cells towards FoxP3 expressing T regulatory cells that secreted IL-10. Gene expression studies revealed that PDGF induced the expression of C-type lectin like receptor member 2, (CLEC-2) receptor on DCs. Furthermore, DCs transfected with CLEC-2 induced T regulatory cells in DC-T cell co-culture. CLEC-2 is naturally expressed on platelets. Therefore, to confirm whether CLEC-2 is responsible for inducing the T regulatory cells, T cells were cultured with either CLEC-2 expressing platelets or soluble CLEC-2. Both conditions resulted in the induction of regulatory T cells. The generation of T regulatory cells was probably due to the binding of CLEC-2 with its ligand podoplanin on T cells, since crosslinking of podoplanin on the T cells also resulted in the induction of T regulatory cells. These data demonstrate that PDGF upregulates the expression of CLEC-2 on cells to induce T regulatory cells.     

5-氟尿嘧啶、长春新碱与去甲二氢愈创木酸配伍对人恶性胶质瘤细胞株的作用

 目的 观察体外应用去甲二氢愈创木酸（ＮＤＧＡ）、予氟尿嘧啶（５－Ｆｕ）、长春新碱（ＶＣＲ）三种药物单用或合用对人恶性胶质瘤细胞系 ＳＨＧ－４４细胞的作用，并探讨其作用的可能机制。方法用 ＭＴＴ法检测药物作用效应，用免疫组化染色法检测细胞 ｃｙｃｌｉｎ Ｄ１基因的蛋白表达情况。结果 （１）三种药物单用及两药合用时随着药物浓度的增加其抗肿瘤效应也增加，且两药合用时药物的效应增强；（２）先给ＮＤＧＡ ２４ｈ后再给 ５－Ｆｕ或 ＶＣＲ，与同时给药对该细胞的抗肿瘤效应无显著差异（Ｐ＞０．０５），而先给 ５－Ｆｕ或 ＶＣＲ ２４ｈ后再给 ＮＤＧＡ，与同时给药对细胞的抗肿瘤效应有显著差异（Ｐ＜０．０５）；（３）免疫组化染色结果表明，与对照组相比，ＮＤＧＡ处理后该细胞 ｃｙｃｌｉｎ Ｄ１基因的蛋白表达明显降低。结论 ＮＤＧＡ与 ５Ｆｕ或 ＶＣＲ间有协同作用，且这种作用可能与 ＮＤＧＡ降低细胞 ｃｙｃｌｉｎ Ｄ１基因的蛋白表达有关。     

Effects of presensitization in vivo on cell-mediated responses to embryonic antigens in vitro

Lymphoid cells specifically reactive with antigens shared by rat bowel carcinomas and mid-term embryo cells were generated by in vitro culture on monolayers of embryo cells. Spleen cells from WF females were cultured for 5 days on monolayers of syngeneic embryo cells or adult cells and assayed for cytotoxic activity on syngeneic embryo, bowel carcinoma, or adult fibroblast target cells in microcytotoxicity and 51Cr-release assays. After culture on embryo monolayers, spleen cells were cytotoxic for embryo and tumor but not for adult fibroblast target cells. Enhanced cytotoxicity was recorded when the spleen cells were cultured from females after interstrain (WF X BN) pregnancy rather than from virgin females. In contrast, previous intrastrain (WF X WF) pregnancy appeared to depress the generation of spleen cells cytotoxic for target cells bearing embryonic antigens.     

肺癌肿瘤干细胞分离鉴定及进展

本文综述肺癌肿瘤干细胞分离鉴定及相关研究进展。肺癌肿瘤干细胞分离主要有依细胞表型特征和生物学特性而建立的两大类方法,ＣＤ１３３、ＡＢＣＧ２、ＢＭＩ-１等可作为分选肺癌肿瘤干细胞细胞标志物。侧群（ｓｉｄｅｐｏｐｕｌａｔｉｏｎ,ＳＰ）细胞分选法可用于未知表面标志物的肺癌肿瘤干细胞分离。肺癌肿瘤干细胞的鉴定分为体外实验和动物致瘤力实验。微流控芯片技术未来可用于肺癌肿瘤干细胞分离鉴定。     

Paracrine interactions between mesothelial and colon-carcinoma cells in a rat model

This study used a co-culture system with Transwell tissue-culture inserts to investigate the role of primary cultures of rat peritoneal mesothelial cells on the proliferation of rat colon-carcinoma cells (CC531 cells). Mesothelial cells significantly inhibited the growth of CC531 cells, while, conversely, CC531 cells stimulated the growth of mesothelial cells. Receptor-binding studies demonstrated the presence of high-affinity IGF-I receptors on the mesothelial and CC531 cells. Both cell types also produced IGF-I, as measured by radioimmunoassay. IGF-I stimulated DNA synthesis in mesothelial cells, but had no effect on the growth of CC531 cells. In co-culture, it was found that IGF-I potentiated the inhibitory effect of mesothelial cells on CC531 cells. The effect of IGF-I on mesothelial-cell proliferation was additive to the stimulatory effect of CC531 cells. TGF-β had no effect on the growth of the CC531 cells, suggesting that this growth (-inhibitory) factor is not involved in the inhibitory effect of mesothelial cells on CC531 cell growth. The study provides evidence for the existence of a paracrine loop between mesothelial and colon-carcinoma cells, giving more insight into the basic cellular mechanisms that may modulate the growth of intraperitoneal colon carcinoma. Inhibition of CC531-cell proliferation by rat mesothelial cells might explain the earlier finding that tumour cells grow poorly in a surgically uncompromised abdomen. Int. J. Cancer 73:885–890, 1997. © 1997 Wiley-Liss, Inc.     

间充质干细胞与肿瘤关系的研究进展

间充质干细胞是一类具有高度自我更新能力和多向分化潜能的组织干细胞,已被视为一种＂种子细胞＂。近年来针对间充质干细胞对肿瘤生长发展的影响已进行了大量的研究,但其对肿瘤的确切作用还未完全清楚,本文就其对肿瘤生长的影响及其相关机制作一简要综述。     

Experimental study of the prophylactic and therapeutic effects of venin on metastasis and recurrence of liver cancer

Objective To study the inhibitory effect of venin on adhesion and invasive ability of SMMC-7721 ceils and to examine the prophylactic and therapeutic effect of venin on liver cancer metastasis and recurrence after hepatectomy. Methods The blocking effect of venin on the intercellular adhesive molecule (ICAM-1) of 7721 cells was analyzed by irnmunofluorescence How cytometry. The influence of venin on the invasive ability of 7721 cells was observed by cell-migration experimentation and detachment of 7721 cells attached to fibronectin (FM), and the influence of venin on adhesion of 7721 cells to FN by the MTT method, 7721 ceils to 7721 cells, 7721 cells lo lymphocytes, and 7721 cells to endoihelial cells by a cellular adhesion test. The preventive and therapeutic eftect of venin on metastasis and recurrence of a liver cancer model was observed in nude mice alter hepatectomy. Results The expression of ICAM-1 in the venin-treated group was significantly lower than that in the untreated group. Venin could not inhibit the invasive ability of 7721 cells, and could not exfoliate the 7721 cells adhered to FN. It could inhibit the adhesion between 7721 cells and 7721 cells, and between 7721 and endothehal cells, but could not inhibit the adhesion between 7721 and lymphocytes. The nude mice treated with venin had less intrahepatic or extrahepatic metastases and recurrences after hepatectomy. Conclusion Venin can inhibit the adhesive ability of SMMC -7721 cells and can also prevent and treat the metastasis and recurrence of liver cancer in nude mice after hepalectomy.     

CD1d expression level in tumor cells is an important determinant for anti-tumor immunity by natural killer T cells

Invariant natural killer T (iNKT) cells are thought to regulate anti-tumor immunity. Human iNKT (i.e. Valpha24+ NKT) cells have been reported to recognize CD1d on target cells and show cytotoxicity directly on the target cells in vitro. However, the anti-tumor effect of mouse iNKT (i.e. Valpha14+ NKT) cells has been repeatedly reported to be dependent on the activity of natural killer (NK) cells via interferon-gamma, with no evidence of direct cytotoxicity. In the present study, we report that in vitro cytolysis of EL-4 mouse lymphoblastic lymphoma cells by Valpha24+ NKT cells and in vivo eradication of these cells are both dependent on the level of CD1d expression on the tumor cell surface. These observations possibly suggest that direct cytotoxicity of tumor cells by iNKT cells is common to both humans and mice, and that the high expression level of CD1d may be a predictor whether the tumor is a good target of iNKT cells.