唾液酸化的路易斯寡糖-X抗原在结直肠癌细胞系LoVo、HT29中的表达及其与转移潜能的相关性

目的　探讨唾液酸化的路易斯寡糖 X (SLeX)抗原的表达与结直肠癌转移潜能的相关性。方法　采用原位分子杂交技术 ,检测结直肠癌细胞表面SLeX抗原合成酶———α1,3岩藻糖转移酶 (α1,3Fuc T)在高、低转移性结直肠癌细胞系LoVo、HT2 9细胞内的mRNA水平的表达 ,并应用免疫组化LSAB法、流式细胞仪 ,从定性、定量两个方面直接检测SLeX抗原在LoVo、HT2 9细胞内蛋白水平的表达。结果　高转移能力的LoVo细胞SLeX抗原呈高水平表达 ( 32 .8± 10 .9,P <0 .0 5 ) ,SLeX抗原合成酶α1,3Fuc TmRNA也呈高水平表达 ( 2 1.2± 7.7,P <0 .0 5 ) ;低转移能力的HT2 9细胞低水平表达SLeX抗原 ( 2 1.9± 8.8) ,并低水平表达SLeX抗原合成酶α1,3Fuc TmRNA( 10 .8± 5 .2 )。结论　SLeX抗原表达与结直肠癌细胞转移潜能密切相关     

CA—50,CEA联合检测对结,直肠癌手术前后的疗效观察

我们对64例结、直肠癌患者进行血清CA 50、CEA测定,并对其手术治疗前、后进行动态观察。现将结果报告如下。 对象和方法 一、对象: (一)正常组:33例来自本院职工体检人员及部分献血者。 (二)恶性肿瘤病人:共64例,为经X线胸片、B超、CT,胃镜、病理、血清学等检查诊断明确的住院病人。 二、方法: (一)标本:血清为常规肘静脉采血后的分离血清,标本均4～8℃贮存备用。 (二)血清CA-50 IRMA试剂盒由中国医学科学院肿瘤所提供。CEA RIA法试剂盒由上海放射免疫分析技术研究所提供,操作均按说明书进行。采用LKB1282Compugamma测定仪。     

肝细胞癌唾液酸化的路易斯寡糖-X抗原和E-上皮钙粘蛋白的表达及其意义

探讨唾液酸化的路易斯寡糖 X抗原 (sialylLewis X ,SLeX)和E 上皮钙粘蛋白 (E cadherin ,ED)表达与肝细胞癌(HCC)转移和预后的关系。应用免疫组织化学方法 ,检测分析 110例HCC组织中SLeX及ED蛋白表达 ,结合随访资料分析。结果显示 ,在HCC中 ,SLeX和ED阳性表达率分别为 39 1%和 4 4 5 %。SLeX阳性表达的HCC转移率高(P <0 0 5 )、分化程度和患者 >5年生存率低 (P <0 0 5 ) ;ED阳性表达的HCC转移率低 (P <0 0 5 )、分化程度和患者>5年生存率高 (P <0 0 5 )。SLeX表达与ED表达呈负相关 (r =- 0 5 3,P <0 0 0 1)。SLeX和ED表达与HCC转移和患者生存期密切相关 ,检测SLeX和ED蛋白的表达可作为判断HCC预后的参考指标     

结直肠癌相关抗原的纯化及分析

<正>肿瘤细胞相关抗原对肿瘤临床诊断及预后监测具有重要意义.近年来应用单克隆抗体技术,鉴定出许多肿瘤相关抗原.目前在临床上应用的结直肠癌标志物除CEA外,还有CA19-9、结直肠癌相关抗原(CCA)等.本文采用鼠抗人结直肠癌单克隆抗体纯化结肠癌组织中的CCA,并对此抗原作初步分析.1 材料与方法1.1 可溶性抗原的制备 人结肠癌组织用生理盐水洗净.将肿瘤组织剪成小块,加入0.01mol/LpH7.2(含3mol/L氯化钾)磷酸缓冲液,制成匀浆.4℃搅拌14h.10000r/min离心30min,上清液用上述缓冲液透析48h.将此提取液边搅拌边滴加3mol/L 高氯酸,至其最终浓度成0.6mol/L,4℃搅拌30min,10000r/min离心30min,上清液透析,稍作浓缩.测定蛋白浓度为2.5×10~2mg/ml.1.2 单抗亲和层析柱的制备和亲和纯化CCA将抗CCA单克隆抗体腹水(IgGl,中国科学院细胞生物研究所提供)经二次盐析后,用pH8.3 0.1mol/L 碳酸缓冲液透析平衡.每毫升 CNBr活化Sepharose 4B凝胶加20mg抗体蛋白混合.室温倒转反应2h,继用1mol/L.pH7乙醇胺封闭2h,制成单抗亲和层析柱.最后用0.15mol/L pH7.2磷酸缓冲液平衡,装柱.将上述高氨酸提取液上亲和层析柱.解高用0.2mol/L pH2.5甘氨酸-HCI缓冲液,收集并及时中和所得抗原蛋白液.1.3 免疫酶法测定 取市售的CCA试剂盒(中科院及本     

结,直肠癌细胞核DNA含量的MSP分析研究

用显微分光光度计检测64例结、直肠癌及其对照肠粘膜细胞核DNA含量,并行细胞核DNA含量分布直方图型分析。淋巴细胞和对照肠粘膜细胞核DNA含量直方图属Ⅰ型。64例结、直肠癌中,56例(87.50%)DNA直方图为Ⅱ～Ⅳ型。与对照肠粘膜细胞比较,结、直肠癌细胞核平均DNA含量、平均核面积和>2C DNA含量的细胞百分率以及各项数值的标准差均明显增加。肠癌细胞核DNA含量直方图与Dukes’分期和分化程度无关。     

结,直肠癌组织中PNA,PHA及CEA的免疫组化研究

用生物素标记的花生凝集素(PNA)、菜豆凝集素(PHA)及抗CEA抗体对良恶性结、直肠组织作组化染色研究,发现:PNA主要标记结、直肠癌(21/25);PHA主要与正常结、直肠杯状细胞粘液相结合(21/22);而癌旁粘膜的结合情况介于二者之间。上述凝集素能显示细胞糖结合物糖化的紊乱,因而有助于结肠癌的诊断。抗CEA抗体可帮助临床决定哪些病人需定期随访检查血中CEA水平,以便监测复发与转移。     

胃和结肠癌组织u—PAR,C—erbB—2和t—PA抗原表达的意义

本文以抗配体抗免疫组化法检测了３５例胃癌和１９例结肠癌组织中尿型纤溶酶原活化素受体（ｕ－ＰＡＲ）同时也测定了癌基因蛋白Ｃ－ｅｒｂＢ－２表达和组织型纤溶酶原活化素（ｔ－ＰＡ）抗原分布，结果表明ｕ－ＰＡＲ阳性表达多见于组织中散在肿瘤细胞表面，胃和结肠癌组织中ｕ－ＰＡＲ阳性表达率分别为２８．６％和３６．８％，Ｃ－ｅｒｂＢ－２分别为３４．３％和４２．１％，这两项指标在浸润型的阳性表达率均显著高于膨胀型（Ｐ     

结直肠癌c—erbB—2,EGFR表达与肿瘤生物学行为的关系

目的：探讨结直肠癌ｃ－ｅｒｂＢ－２及表皮生长因子受体（ＥＧＦＲ）共同表达对其生物学行为及预后的影响。方法：应用免疫组织化学方法检测６９例结直肠癌中ｃ－ｅｒｂＢ－２及ＥＧＦＲ的表达情况。结果：ｃ－ｅｒｂＢ－２和ＥＧＦＲ共同表达时，肿癌细胞增殖指数高，呈浸润性生长方式的较多，病人５年生存率较低。结论：ｃ－ｅｒｂＢ－２和ＥＧＦＲ可能有协同作用，同时检验两者的表达情况可能为临床判断病人预后提供较可靠的依据     

nm23—H1,c—erbB—2基因表达与结直肠癌转移的？…

目的：探讨ｎｍ２３－Ｈ１，ｃ－ｅｒｂＢ－２基因表达与结直肠癌转移的关系，方法：采用免疫组化ＳＡＢＣ法，检测了８５例结直肠癌中ｎｍ２３－Ｈ１，ｃ－ｅｒｂＢ－２基因的蛋白产物，结果：ＤｕｋｅｓＤ期结直场癌中ｎｍ２３－Ｈ１基因表达显著低于ＤｕｋｅｓＡ，Ｂ，Ｃ各期（Ｐ〈０．０１），ｃ－ｅｒｂＢ－２基因表达则显著高于Ｄｕｋｅｓ，Ａ，Ｂ，Ｃ各期（Ｐ〈０．０１），两基因在ＤｕｋｅｓＡ，Ｂ，Ｃ各期间的阳性表达差异     

具有相同遗传背景的人结肠癌细胞系生物学特性的鉴定

目的 对三种取自不同转移部位的结肠癌细胞系进行生物学特性的鉴定.方法 从体内成瘤性、原位移植后的自发性转移能力,以及体外生长、克隆形成率、粘附、运动、侵袭能力等方面,探讨具有相同遗传背景的SW480、SW620以及SW480/M5三种细胞系生物学特性的差异.结果 体内实验证明SW480具有多器官转移的潜能,SW620只具有淋巴结转移的能力,SW480/M5只具有肝脏转移的能力,SW480/M5的皮下瘤生长速度最快,其次是SW620细胞;体外实验证明W48/M5和SW620的体外生长、克隆形成能力均强于SW480细胞,而两者对纤维黏连蛋白的粘附能力较SW480细胞弱,SW480/M5的运动及侵袭能力强于SW480及SW620细胞.结论 与SW480细胞相比,SW480/M5和SW620细胞具有一定的器官亲和力,三者的遗传背景一致,是研究结肠癌晚期演进的遗传改变的重要资源.     

Cytoskeletal stiffness, friction, and fluidity of cancer cell lines with different metastatic potential

We quantified mechanical properties of cancer cells differing in metastatic potential. These cells included normal and H-ras-transformed NIH3T3 fibroblast cells, normal and oncoprotein-overexpressing MCF10A breast cancer cells, and weakly and strongly metastatic cancer cell line pairs originating from human cancers of the skin (A375P and A375SM cells), kidney (SN12C and SN12PM6 cells), prostate (PC3M and PC3MLN4 cells), and bladder (253J and 253JB5 cells). Using magnetic twisting cytometry, cytoskeletal stiffness (g′) and internal friction (g″) were measured over a wide frequency range. The dependencies of g′ and g″ upon frequency were used to determine the power law exponent x which is a direct measure of cytoskeletal fluidity and quantifies where the cytoskeleton resides along the spectrum of solid-like (x = 1) to fluid-like (x = 2) states. Cytoskeletal fluidity x increased following transformation by H-ras oncogene expression in NIH3T3 cells, overexpression of ErbB2 and 14-3-3-ζ in MCF10A cells, and implantation and growth of PC3M and 253J cells in the prostate and bladder, respectively. Each of these perturbations that had previously been shown to enhance cancer cell motility and invasion are shown here to shift the cytoskeleton towards a more fluid-like state. In contrast, strongly metastatic A375SM and SN12PM6 cells that disseminate by lodging in the microcirculation of peripheral organs had smaller x than did their weakly metastatic cell line pairs A375P and SN12C, respectively. Thus, enhanced hematological dissemination was associated with decreased x and a shift towards a more solid-like cytoskeleton. Taken together, these results are consistent with the notion that adaptations known to enhance metastatic ability in cancer cell lines define a spectrum of fluid-like versus solid-like states, and the position of the cancer cell within this spectrum may be a determinant of cancer progression.     

Ki-67表达与结直肠癌Dukes分期关系

目的检测ki-67在正常结直肠组织和结直肠癌中的表达情况,探讨其表达与结直肠癌Dukes分期关系,为临床结直肠癌的诊疗判断提供参考。方法收集临床病理资料完整的结直肠癌54例,正常结直肠黏膜组织28例。应用免疫组化法(SP法)检测正常结直肠组织和结直肠癌中ki-67蛋白的表达。结果Ki-67蛋白在结直肠癌及正常结直肠黏膜中的表达率分别为85.19%和10.71%,在结直肠癌组织中的表达高于正常结直肠黏膜的表达,差异有统计学意义(P<0.01)。ki-67蛋白的表达和Dukes分期无相关性,差异无统计学意义(P>0.05)。结论ki-67蛋白过表达与结直肠癌的发生发展密切相关,但不能作为结直肠癌预后的参考指标。     

The relationship of BRMS1 and RhoGDI2 gene expression to metastatic potential in lineage related human bladder cancer cell lines

We have recently characterized a human bladder cancer cell line T24 and a more aggressive lineage related variant of it, T24T. To gain further insights, we have studied their metastatic ability in an in vivo model system. Results show that T24 forms significantly fewer [4/12 (1/11) mice had metastases with 1-2 lesions/mouse] metastasis in SCID/bg mice than T24T [14/14 (6/6) mice had metastases with a mean of 24-28 lesions/mouse]. To begin exploring the mechanisms underlying this difference, we evaluated the mRNA and protein expression levels of metastasis-suppressor genes, known to be important in the progression of other cancers, in our model of bladder cancer progression. A higher mRNA expression of BRMS1, a metastasis suppressor in breast cancer, was observed in T24 cells. In addition, RhoGDI2 mRNA expression was only observed in T24 when compared to T24T, suggesting that Rho activation might play a significant role in the metastatic cascade. However, a basal level mRNA expression of KISS1, described as metastasis suppressor in melanoma and breast, was observed in both the lines and had slightly higher expression in T24T. No difference of Nm23-H1, KAI1, MKK4/SEK1 and E-Cadherin protein levels were noted between these two lines. In summary, it appears that the T24/T24T paired cell lines constitute a useful model for the study of human bladder cancer metastasis that will allow both the discovery and mechanistic evaluation of genes potentially involved in this process. This revised version was published online in July 2006 with corrections to the Cover Date.     

不同转移潜能肿瘤细胞胸腺素β10表达水平及微丝结构的差异性研究

目的 研究胸腺素β10(Tlaymosinβ10,Tβ10)表达水平与人类肿瘤转移潜能的相关性以及细胞内微丝骨架的变化差异。方法 利用Northern-blot和细胞爬片免疫组织化学的方法检测4组9种具有不同转移潜能的人类肿瘤细胞系中Tβ10 mRNA和蛋白的表达水平;通过组织化学的方法显示细胞内微丝骨架的变化。结果 Tβ10的mRNA和蛋白表达水平在高转移潜能的人类肺癌、黑色素瘤和乳腺癌细胞中高于相应的不／低转移肿瘤细胞;高转移性癌与不／低转移癌细胞比较,前者细胞内肌动蛋白多聚体减少,微丝骨架杂乱、模糊,而低转移癌系微丝骨架粗大、结构清晰、排列整齐。结论 在所研究的人类肿瘤中,Tβ10 mRNA和蛋白表达水平与肿瘤的转移能力呈正相关性;肿瘤转移潜能的增高伴有肌动蛋白多聚体的丢失和微丝结构的解聚,微丝骨架的这种变化与Tβ10的表达增高具有相关性。     

结直肠癌细胞中脾酪氨酸激酶基因甲基化状态和表达的关系

目的：探讨结直肠癌细胞中脾酪氨酸激酶基因甲基化和表达的关系。方法：应用亚硫酸盐修饰测序、甲基化特异性聚合酶链反应和蛋白印迹技术检测结直肠癌细胞脾酪氨酸激酶的甲基化状态以及表达情况；荧光素酶报告分析法研究启动子区域CpG岛的甲基化与启动子活性的关系；甲基化转移酶抑制剂处理脾酪氨酸激酶甲基化失表达的结直肠癌细胞株，观察处理前后细胞内脾酪氨酸激酶基因甲基化状态和表达情况。结果：（1）23个结直肠癌细胞中，9个细胞启动子发生甲基化而失去蛋白质表达；其余则正常表达，甲基化发生率为39.2％；（2）9个甲基化的细胞中，7个存在微卫星不稳定；而14个未发生甲基化的细胞中，仅有4个存在微卫星不稳定。二者之间的差异显著（P<0.05）；（3）脾酪氨酸激酶启动子全长和未甲基化启动子荧光素酶的活性分别是甲基化组的4.5和4.7倍；5-Aza-CdR可恢复甲基化启动子的活性；（4）5-Aza-CdR可去甲基化而使脾酪氨酸激酶基因重新表达，而且具有时间依从性。结论：结直肠癌细胞中，启动子区域的甲基化导致Syk基因丧失表达，5-Aza-CdR可以去甲基化而恢复脾酪氨酸激酶基因的表达。     

Modulation of cell growth and PPARgamma expression in human colorectal cancer cell lines by ciglitazone

Studies have shown that ligand activation of peroxisome proliferator-activated receptor gamma (PPARgamma) can induce differentiation and inhibit proliferation of several cancer cells. The present study was performed to investigate the effects of the PPARgamma ligand, ciglitazone, and the involvement of PPARgamma in modulating the growth of human colorectal cancer cells. Lactate dehydrogenase release assay showed that ciglitazone potently inhibited HT-29 (well-differentiated) and COLO-205 (poorly differentiated) colorectal adenocarcinoma cell growth. Measurement of apoptosis by flow cytometry using a fluorescein-conjugated monoclonal antibody against cytokeratin 18 revealed a high induction of apoptosis by ciglitazone in a time-dependent fashion. The expression of PPARgamma1 but not PPARgamma2 mRNA was significantly downregulated as measured by real-time quantitative PCR, and the PPARgamma protein levels were decreased as determined by Western blot analysis. We conclude that ciglitazone treatment suppressed colon cancer cell growth via induction of apoptosis. However, the anticancer effects of ciglitazone may not depend solely on PPARgamma activation.     

Aurora-B expression and its correlation with cell proliferation and metastasis in oral cancer

Aurora-B kinase is a chromosomal passenger protein and is essential for chromosome segregation and cytokinesis. Aurora-B overexpression in various cancer cells induces chromosomal number instability to produce multinuclearity and relates to metastasis. Here, we examined the expression of Aurora-B in oral squamous cell carcinoma (OSCC) to elucidate the relationship between Aurora-B expression and clinico-pathological findings by immunohistochemistry. Aurora-B expression was observed in normal oral squamous epithelia and OSCC cases, but the number of positive cells was significantly higher in OSCC than in normal squamous epithelium (p < 0.01). The labeling index of Aurora-B was significantly correlated with lymph node metastasis (p < 0.01) and histological grades of differentiation (p < 0.01). We also compared Aurora-B expression with Ki-67 expression and a positive correlation was found (p < 0.0001). Moreover, Aurora-B expression is significantly more frequent in multinuclear tumor cells than in total tumor cells. In summary, we found that Aurora-B expression was well correlated with cell proliferation, induction of multinuclear cells, histological differentiation, and metastasis in OSCC. These findings suggest that Aurora-B may be involved in tumor progression and that Aurora-B can be a new diagnostic and therapeutic target for OSCC.     

BRI基因在一对转移能力不同的肺腺癌细胞系中的序列分析与表达研究

目的 确认淀粉样纤维蛋白基因(amyloid fibrils，BRI)基因在l对同源但转移能力不同的肺腺癌细胞系AGZY83-a和Anip973中的序列并分析其表达。方法 采用测序技术，Northern印迹杂交。G显带后荧光原位杂交分析BRI基因在肺腺癌细胞系的序列与表达。结果 BRI基因在高转移肺腺癌细胞系Anip973中高表达，在其低转移母系AGZY83-a中低表达，两细胞系BRI基因染色体定位区均存在断裂重排。该基因染色体定位区在Anip973中出现扩增。已知BRI基因的-116bp～-5bp处碱基序列和-115bp～-5bp处碱基序列在AGZY83-a和Anip973中分别突变为CTCAGCAGCCCGC和TCAGCCGC。结论 BRI基因在转移能力不同的肺腺癌细胞系差异表达与该基因的染色体定位区域的断裂重排无关，与该基因染色体定位区拷贝数增加及5′非翻译区存在不同的突变可能相关。     

Aberrant Cosmc genes result in Tn antigen expression in human colorectal carcinoma cell line HT-29

The Tn antigen, which arises from mutation in the Cosmc gene is one of the most common tumor associated carbohydrate antigens. Cosmc resides in X24 encoded by a single gene and functions as a specific molecular chaperone for T-synthase. While the Tn antigen cannot be detected in normal cells, Cosmc mutations inactivate T-synthase and consequently result in Tn antigen expression within certain cancers. In addition to this Cosmc mutation-induced expression, the Tn antigen is also expressed in such cell lines as Jurkat T, LSC and LS174T. Whether the Cosmc mutation is present in the colon cancer cell line HT-29 is still unclear. Here, we isolate HT-29-Tn⁺ cells from HT-29 cells derived from a female colon cancer patient. These HT-29-Tn⁺ cells show a loss of the Cosmc gene coding sequence (CDS) leading to an absence of T-synthase activity and Tn antigen expression. Additionally, almost no methylation of Cosmc CpG islands was detected in HT-29-Tn⁺ as well as in HT-29-Tn^- and Tn^- tumor cells from male patients. In contrast, the methylation frequency of CpG island of Cosmc in normal female cells was ~50%. Only one active allele of Cosmc existed in HT-29-Tn⁺ and HT-29-Tn^- cells as based upon detection of SNP sites. These results indicate that Tn antigens expression and T-synthase inactivity in HT-29-Tn⁺ cells can be related to the absence of CDS in Cosmc active alleles, while an inactive allele deletion of Cosmc in HT-29 cells has no influence on Cosmc function.