Expression of Fas Antigen and Its Mediation of Apoptosis in Human Gastric Cancer Cell Lines

Fas, a member of the tumor necrosis factor receptor/nerve growth factor receptor family, induces apoptosis by crosslinking with Fas ligand or anti-Fas antibody in a variety of cultured cells. We examined the expression of Fas antigen and its mediation of apoptosis in six human gastric carcinoma cell lines. Flow cytometric analysis and western blotting revealed relatively high expression of Fas antigen in MKN-74 (wild-type p53 gene) and MKN-45 (wild-type), followed by MKN-1 (mutated), MKN-7 (mutated) and KATO-III (deleted). MKN-28 (mutated) showed minimal expression of the antigen. The expression was apparently enhanced by interferon-γ, except for MKN-1 and MKN-28. Anti-Fas antibody (100 ng/ml) induced nuclear fragmentation characteristic of apoptosis. Apoptosis occurred in a delayed fashion and the apoptotic index at 72 h was approximately 60% in MKN-74, 35% in MKN-45, and 20% in MKN-1 and KATO-III. A DNA ladder was noted in MKN-74 at 72 h. Expression levels of P53 and P21^Wafl did not change for up to 48 h in MKN-74. The biological effects did not correlate with endogenous Bcl-2 expression. These results indicated that a) Fas antigen is variably expressed in human cultured gastric carcinoma cells, b) the protein transduces an apoptotic signal which leads to delayed cell death, and c) susceptibility to the antibody correlates well with the expression level of Fas antigen.     

Expression of C-terminal src Kinase in Human Colorectal Cancer Cell Lines

C-terminal src kinase (CSK) is a cytoplasmic protein which decreasesactivities of the Src family protein-tyrosine kinases. We produceda polyclonal antibody specific for CSK and analyzed the expressionof CSK by immunoblotting in two human colorectal normal celllines and eighteen cancer cell lines. CSK was detected in boththe two normal and all the eighteen cancer cell lines. The expressionof CSK obtained from human colorectal cancer cell lines wasgreater than that from human colorectal normal cell lines inmost cases. The elevated expression of CSK in human colorectalcancer cell lines appeared to correspond to the elevated p60^c-src(c-Src) and p62^c-yes (c-Yes) protein-tyrosine kinase activitiesfound in other studies. Thus, CSK may not have an anti-oncogenicrole to play through the negative regulation of Src family kinasesin colorectal carcinogenesis     

Expression and Roles of Heat Shock Proteins in Human Breast Cancer

Heat shock proteins (hsps) are thought to play important roles in the cell cycle and various processes of carcinogenesis. Therefore, we evaluated the expression of hsps, mainly hsp90 and hsp70, in human breast cancer tissues. Hsp90α mRNA was expressed at much higher levels in the cancerous tissue than in the non-cancerous tissue. In addition, a close correlation between hsp90α mRNA expression and the proliferating-cell-nuclear-antigen labeling index (PCNA LI) was observed for the cancerous tissue. These findings suggest that increased expression of the hsp90α isoform may play a role in cell proliferation. On the other hand, hsp90β mRNA expression was significantly higher in poorly differentiated carcinomas than in well differentiated carcinomas of the breast. The intracellular localization of hsp70 was consistent with that of ubiquitin. In specimens showing hsp70 in the nucleus, the PCNA LI was significantly high, Hsc73 mRNA, a member of the hsp70 family, was also expressed at higher levels in cancerous tissues associated with a high PCNA LI than in non-cancerous tissues. These results suggest that hsp90α may play a role in cancer cell proliferation and that hsp90β may contribute to cell differentiation and structural constitution. In addition, hsp70, especially hsc73, is related to ubiquitin and seems to be a marker for cancer proliferation.     

The Difference of Gene Expression Prof ile in Human Large Cell Lung Cancer Cell Lines with Different Metastatic Potential

Background and Objective Lung cancer is the most lethal malignangy that threatens human heath and lives nowadays in the world, and meanwhile is also the one with worst clinical     

细胞周期抑制蛋白p16、p27异常表达与胃癌生物学行为的关系

目的：探索细胞周期抑制蛋白p16、p27异常表达与胃癌生物学行为的关系。方法：免疫组织化学方法（S－P法）检测胃癌组织中p16,p27的表达,结合临床病理资料和随访资料进行回顾性研究,并作出评价。结果：p16表达降低与胃癌分期增加、淋巴结转移及不良预后显著相关。p27表达降低与淋巴结转移、浆膜浸润、晚期胃癌、弥漫型胃癌及不良预后显著相关。结论;p16,p27异常表达与胃癌不良的生物学行为及预后有关,可作为预后指标,有利于个体化治疗及提高胃癌的疗效。     

Alteration of Gene Expression Profile and Signal Pathway Induced by Methylseleninic Acid in Human Lung Cancer Cell Lines

Objective and Methods Lung cancer has a fastest growing rate of morbidity and mortality among malignant tumors and poses a great threat to the human health. Chemotherapy, as one of     

膀胱癌组织中EGFr和p21蛋白的表达评价

应用ｐ２１和表皮生长因子受体（ＥＧＦｒ）单克隆抗体，通过免疫组织化学方法检测正常膀胱组织和膀胱癌组织中ｒａｓ癌基因产物ｐ２１和ＥＧＦｒ的表达状况。结果显示：５例正常膀胱组织中未发现ＥＧＦｒ和ｐ２１阳性表达，阳性表达的ＥＧＦｒ和ｐ２１均定位于肿瘤的细胞膜上。５８例膀胱癌中ＥＧＦｒ和ｐ２１阳性表达率分别为５８．６％（３４／５８）和６２．１％（３６／５８）．并且ＥＧＦｒ和ｐ２１阳性表达率与膀胱癌的病理分级、临床分期和患者术后生存期有关。提示ＥＧＦｒ和ｐ２１阳性表达在膀胱癌发生和发展中起着重要作用，可判别膀胱癌恶性程度和预测膀胱癌的预后。     

经典瞬时受体电位通道蛋白在人非小细胞肺癌组织中的表达

背景与目的 经典瞬时受体电位(transient receptor potential canonical,TRPC)通道蛋白是一种非选择性阳离子通道蛋白家族,主要位于细胞膜表面,对钙离子具有通透性.研究认为,TRPC可能构成钙池操纵性钙通道(store-operated calcium channels,SOCC)并介导钙池操纵性钙内流(store-operated calcium entry,SOCE),从而参与细胞的增殖、迁移、基因转录等生命活动.本研究检测非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中TRPC mRNA及蛋白质的表达情况,初步探讨TRPC与NSCLC的可能关系.方法 建立TRPC1-7等7个家族成员的荧光定量PCR检测方法,对24例NSCLC患者的肿瘤组织进行了TRPC mRNA的定量检测,并通过蛋白质免疫印迹法对TRPC在蛋白质水平的表达进行了验证.结果 在NSCLC患者癌组织检测到TRPC1、TRPC3、TRPC4和TRPC6 mRNA的表达,未检测到TRPC2、TRPC5和TRPC7 mRNA的表达.肺癌组织中TRPC表达丰度为:TRPC1≈TRPC6＞TRPC3＞TRPC4.蛋白质免疫印迹证实了非小细胞肺癌组织中TRPC1、TRPC3、TRPC4和TRPC6在蛋白质水平的表达.结论 非小细胞肺癌组织在mRNA和蛋白质水平均表达TRPC1、TRPC3、TRPC4和TRPC6,其中主要表达TRPC1和TRPC6,它们在构成肺癌细胞中SOCC、介导产生SOCE中的作用有待进一步研究.     

增殖细胞核抗原与卵巢癌细胞系化学敏感性的关系

目的　探讨增殖细胞核抗原 (PCNA)与上皮性卵巢癌细胞系化学敏感性的关系。方法　使用顺铂作用于A 2 780及AD 60细胞株。在MTT法测定耐药倍数基础上 ,用ABC检测PCNA表达。结果　MTT测定AD 60的耐药倍数为 2 .4。A 2 780PCNA的表达低于AD 60 ,且随药物浓度的递增染色较耐药组明显变浅。A 2 780、AD 60相同药物浓度作用后PCNA的表达有显著差异 (P <0 .0 1) ;同一细胞株不同药物浓度作用后PCNA表达有显著性差异 (P <0 .0 1)。结论　增殖细胞核抗原表达与上皮性卵巢癌细胞对细胞毒性药物的化学敏感性呈负相关。     

The Expression Levels of KAI1 Gene and Its Mechanism in Human Lung Cancer Cell Lines

Background and Objective Lung cancer is one of the most malignant cancers which is hazarding the people’s health and life in the world. In the past half century, the incidence and mortality of     

GLA对肺癌细胞株E-CD表达及侵袭能力的影响

目的:观察GLA对肺癌细胞株体外生长抑制,E-CD表达及侵袭能力的影响。方法:采用细胞计数、SP免疫组织化学、图像分析及侵袭小室等方法,观察人肺腺癌细胞株A549、小细胞肺癌细胞株NC-H446在GLA、ARTA处理不同时间后细胞生长、E-CD表达及侵袭能力的影响。结果:在GLA干预下,NC-H446、A549两细胞株生长受到抑制,E-CD阳性表达率分别由原来的12%±8%、23%±11%提高到34%±9%、39%±13%,IOD分别由78.5%±16.9%、109.8%±13.2%增加到200.3%±21.7%、229.7%±11.0%,前后相比具有显著差异(P<0.01);穿过基质胶的细胞数分别为126.5±43.61、106.9±55.48,与对照组相比有显著差异(P<0.01);上述情况与ARTA相比,无明显差异(P>0.05)。结论:GLA能明显抑制两细胞株生长,促进E-CD的正常表达,减弱肿瘤细胞的侵袭转移能力。     

功能性Fas 配体在大肠癌细胞的表达及其调控

 目的　研究功能性 Fas配体在大肠癌细胞的表达及其调控。方法　分别采用反转录 -聚合酶链反应和氚释放细胞活性测定技术检测了大肠癌细胞株中 Fas配体 m RNA表达和 Fas依赖的细胞毒活性及其调控。结果　在检测的六株大肠癌细胞中 ,全部表达 Fas配体 m RNA。部分大肠癌细胞具有 Fas依赖的细胞毒活性。其中 DLD- 1细胞活性具有量效关系 ,并且乙酸肉豆寇佛波醇 (PMA)和离子霉素 (ionomycine)刺激能显著增强 DLD- 1的细胞毒活性。结论　至少部分大肠癌细胞表达功能性 Fas配体 ,并可能以此反击机体免疫系统 ,逃避免疫监督。     

miRNA-34 a对人结肠癌细胞 HCT116增殖及侵袭转移的影响

目的：研究微小RNA-34a（microRNA-34a,miR-34a）在人结肠癌细胞株HCT116中的过表达对细胞增殖和侵袭的影响及其机制。方法设计合成并构建携带绿色荧光蛋白（ GFP）的miR-34a真核表达载体,脂质体法稳定转染HCT116细胞。通过Realtime PCR验证转染后miR-34a的表达变化。 MTT法检测转染前后HCT116细胞增殖能力变化, Transwell法检测转染前后HCT116细胞侵袭能力改变,western blot检测目的蛋白的表达。结果 miR-34a转染HCT116细胞后其表达量增加到（7．32±1．34）倍,而HCT116细胞增殖能力也受到显著抑制,下降到（0．49±0．10）;Bcl-2蛋白受到miR-34a表达的抑制,而BAX的表达则受到miR-34a表达的增强;HCT116细胞侵袭能力在miR-34a过表达后显著增强,相应的MMP-2和MMP-9表达也出现显著增强。阴性对照组与空白对照组比较无显著性差异。结论 miR-34a的过表达能够抑制人结肠癌细胞HCT116的增殖及侵袭转移,与调控Bcl-2／BAX和MMP-2和-9蛋白的表达密切相关。     

抗人胃癌3H11人-鼠嵌合抗体的构建及表达

为了降低抗人胃癌鼠单抗3H11的免疫原性,以利于该单抗在临床中的应用。我们构建并表达了3H11的人-鼠嵌合抗体,将3H11的轻、重链可变区基因分别插入到含有人k链及IgGl重链恒定区基因的真核细胞表达载体中,构建了3H11人-鼠嵌合抗体轻、重链表达载体。应用Lipofectin方法先将嵌合轻链表达载体转染到Sp2/0细胞中,经用含霉酚酸的选择培养基筛选及克隆化培养,获得稳定分泌3H11人-鼠嵌合轻链的转染细胞株。再将嵌合重链表达载体转染该细胞系,用含有组氨醇的选择培养基筛选,获得组氨醇抗性细胞株,经亚克隆后得到可稳定分泌人k链和人IgGl的转染细胞系,经ELISA检测该细胞系所分泌的上清含有可与人胃癌细胞系803结合的人IgG抗体活性,RT-PCR结果显示该细胞株有人-鼠嵌合抗体mRNA的转录,证明已获得分泌3H11人-鼠嵌合抗体的细胞系。     

沙立度胺对人乳腺癌细胞增殖及血管内皮生长因子表达的影响

[目的]观察沙立度胺对人乳腺癌细胞系MCF-7、MDA-MB-231细胞增殖以及血管内皮生长因子(VEGF)表达的抑制作用.[方法]应用MTT方法及半定量RT-PCR技术,分别观察沙立度胺对MCF-7和MDA-MB-231细胞增殖及VEGF表达的抑制作用.[结果]沙立度胺在(10～100)μg/ml范围内对MCF-7和MDA-MB-231细胞有明显的抑制增殖作用,并伴随VEGFmRNA表达水平下降;50μg/ml明显抑制两株细胞VEGFmRNA的表达.[结论]沙立度胺在一定浓度范围内能抑制乳腺癌细胞VEGFmRNA的表达,抑制细胞增殖反应.     

High Expression of MICA in Human Kidney Cancer Tissue and Renal Cell Carcinoma Lines

The overall incidence and mortality of renal cell carcinoma (RCC), the most common kidney cancer, aresteadily increasing for reasons that are not fully explained. Our aim was to explore the expression of membraneMHC class I chain-related gene A (mMICA) in human RCC cell lines and tissue specimens, and to determineexpression of soluble MICA (sMICA) in serum of patients with renal cell carcinoma, we used flow cytometry(FCM) and immunohistochemistry as well as an enzyme linked immunosorbent assay (ELISA) . The resultsshowed that percentage of mMICA expression was significantly increased in human kidney cancer tissues andRCC cell lines (786-O and Ketr-3) than that in healthy adults and human embryonic kidney 293 (HEK293)cell line individuality (P<0.05). sMICA content in healthy adults was negative, but in renal cancer patients wassignificantly elevated (P<0.05). Our research showed that high expression of MICA in human kidney cancer,this results show that MICA might serve as potential tumor-associated antigen (TAA) in RCC.     

Induction of p53-independent Apoptosis Associated with G2M Arrest Following DNA Damage in Human Colon Cancer Cell Lines

The tumor suppressor p53 protein induces apoptosis in response to various kinds of DNA damage in normal cells, but it is still unclear whether or not apoptosis induced by DNA damage correlates with the p53 status in tumor cells. We determined the status of p53 by functional analysis of separated alleles in yeast in five human colon cancer cell lines, SW-480, SW-620, DLD-1, COLO320 and LS174T and investigated whether p53 is necessary for apoptosis and cell cycle arrest after treatment of the cells with a DNA-damaging agent, etoposide (VP-16), or γ-irradiation. Of these cell lines, only LS174T expresses a functional p53. Apoptosis was detected in SW-480 and COLO320 cell lines, but not in the other cell lines, including LS174T cell line with a normal p53 function. Furthermore, cell cycle analysis revealed accumulation in the G2M phase preceding induction of apoptosis in SW-480 and COLO320 cells, but not in the other cells. These results suggest that apoptotic induction by DNA damage is not necessarily related to p53 status and that induction of p53-independent apoptosis following DNA damage may correlate with G2M arrest in the cell cycle, at least in the colon cancer cell lines used in this study.     

常用化疗药物对大肠癌细胞系PTEN蛋白表达的影响

目的　探讨常用化疗药物对大肠癌细胞系PTEN蛋白表达的调控作用。方法　采用WesternBlot法 ,检测氟脲嘧啶( 5 Fu)、丝裂霉素及顺铂作用于大肠癌细胞HT 2 96h及 2 4h后PTEN蛋白表达情况。结果　 3种药物作用 6h后 ,与对照组比较 ,PTEN表达量均呈不同程度增高 ,其中以顺铂的上升幅度最大 ,为对照组的 2 .16倍 ,5 Fu为对照组的 1.2 1倍 ,丝裂霉素为对照组的 1.5 0倍。作用 2 4h后 ,PTEN表达量顺铂为对照组的 3 .5 3倍 ,5 Fu为对照组的 1.68倍 ,丝裂霉素为对照组的 2 .2 1倍。且用上述药物作用 6h后 ,大肠癌细胞的生长速度较对照组明显减慢。结论　常用化疗药物可通过上调癌细胞中PTEN表达量来实现其抗癌作用