紫杉醇诱导人肺腺癌多药耐药细胞系的建立及DNA polβ、mdr1、mrp1、GST-πl、rp和topo Ⅱ基因的表达

目的体外建立人肺腺癌细胞多药耐药细胞系,观察其生物学特性并探讨其细胞内DNA polβ和多药耐药基因mdr1、mrp1、GST-πl、rp和topo Ⅱ的表达及意义。方法以紫杉醇为诱导药,采用间歇逐步增加剂量法建立人肺腺癌多药耐药细胞系A549/TXL20;MTT法检测A549/TXL20对紫杉醇、顺铂、长春新碱、5-氟尿嘧啶和丝裂霉素的耐药指数(RF);细胞克隆形成法测定A549/TXL20对紫杉醇的敏感性;高效液相色谱测定细胞内紫杉醇的浓度;RT-PCR法测定人肺腺癌多药耐药细胞系A549/TXL20中DNA polβ、mdr1、GST-π、mrp1l、rp和topo Ⅱ的基因表达。结果①A549/TXL20形态不规则,较亲本细胞略小,核/浆比增加,倍增时间没有明显变化;②A549/TXL20对紫杉醇的RF为19.3,对顺铂的RF为67.4,对长春新碱、5-氟尿嘧啶、丝裂霉素和顺铂等抗癌药物有不同程度的耐药性;③A549/TXL20对紫杉醇的敏感性降低,其紫杉醇的含量较亲本细胞下降;④A549/TXL20细胞中DNA polβ、mdr1、GST-π、mrp1l、rp的基因表达量分别较A549细胞中的表达量明显增加,差异具有统计学意义(P<0.05)。结论建立了相对稳定的人肺腺癌多药耐药细胞系A549/TXL20,其耐药机制可能与mdr1、GST-π、mrp1l、rp和DNA polβ基因的高表达有关。     

L-精氨酸对实验性结肠癌的作用与机理

目的　研究L 精氨酸在Wistar大鼠实验性结肠癌中的作用及其机制。方法　用致癌物1 ,2 二甲基联苯 (DMH)每周 1次皮下注射 2 0周建立大鼠结肠癌模型 ,给予L 精氨酸溶液口服 ,对照组皮下注射EDTA ,大鼠共分为 6组 ,每组 1 8～ 2 0只。 2 2周后处死动物 ,研究肿瘤特性、全血淋巴细胞对有丝分裂素PHA的反应、T细胞亚型 (CD2 ,CD4,CD8)及血浆胃泌素浓度变化。结果　精氨酸可明显降低结肠癌的体积 ,而对结肠腺瘤则无明显差异。早期全程应用精氨酸明显减少侵犯到粘膜下层的腺癌数量 ,刺激淋巴细胞对有丝分裂素的反应并提高T细胞亚型的分布。但精氨酸不改变CD4/CD8比率以及胃泌素的浓度。结论　精氨酸降低实验性结肠癌的生成可能与其非特异性刺激机体淋巴细胞免疫功能有关 ,这种作用在肿瘤发生早期更显著。     

大肠癌及癌旁组织中糖链抗原50的表达

目的探讨糖链抗原50(CA50)在大肠癌及癌旁组织中表达的意义。方法采用免疫组化检测正常大肠黏膜(10例)、大肠癌远旁(40例)、近旁(40例)和癌(40例)组织中CA50的表达。结果CA50阳性表达率按正常大肠黏膜、大肠癌远旁、近旁、癌组织的顺序依次增高,且阳性表达率在4组间有显著性差异(P<0.05)。CA50表达与大肠癌的分化程度、Duke s分期和淋巴结转移有关(P<0.05),与组织学类型无关(P>0.05)。结论CA50的表达对评价大肠癌的生物学特性、判断预后具有重要的临床意义。     

Depression and oxidative damage in TNM stage Ⅲ patients with poorly differentiated gastric adenocarcinoma

Objective To investigate the association between psychological stress and oxidative damage in TNM stage Ⅲ patients with poorly differentiated gastric adenecarcinoma (GA). Methods One hundred and six patients with newly diagnosed poorly differentiated GA were assessed using the Hamilton Depression Rating Scale (HAMD), Zung Self-rating Depression Scale (SDS), Zung Self-rating Anxiety Scale (SAS), Symptom Checklist 90 (SCL-90), activities of daily living (ADL) and other multiple-item qnestionnaires. Oxidative-stress-related parameters in serum and the expression of DNA repair genes were monitored during a pretreatment period. Results The patients were divided into depression and nondepression groups (Groups A and B, respectively) based on a HAMD score cutoff of 20. The mean SDS, SAS, SCL-90, ADL and passive coping scores were higher in Group A, whereas social support and quality of life were lower. Serum total antioxidant capacity, eatalase, superoxide dismutuse concentrations and anti-superoxide anion capacity (A-ASC) were significantly decreased in Group A, whereas serum malondialdehyde (MDA) and 8-hydroxy-deoxyguanosine (8-OHdG) levels were significantly increased. Pearson correlation analysis revealed that depression was pesitively correlated with MDA, SAS, SCL-90 and ADL, but negatively correlated with A-ASC. Furthermore, real-time PCR revealed that the expression levels of hOGG1 and APEX1 were increased in Group A. Conclusion Psychological stress might be related to impaired antioxidant system in patients with GA, and it presents the first evidence of the involvement of oxidative DNA damage in the pathogenesis of depression.     

Ⅰ型胶原蛋白基因在反流性食管炎、Barrett食管及食管腺癌中的表达

目的探讨反流性食管炎、Barrett食管及食管腺癌黏膜组织中Ⅰ型胶原蛋白(COL1A1)基因mRNA的表达水平及意义。方法经胃镜取材,采用荧光定量PCR方法检测反流性食管炎、Barrett食管、食管腺癌组织中COL1A1基因mRNA的表达水平。结果 COL1A1mRNA在正常组和反流性食管炎阴性组食管黏膜中呈低表达(0.154±0.119,0.152±0.105),在Barrett食管与食管腺癌食管黏膜中呈明显的高表达(0.396±0.170,0.726±0.561)。并且在从正常→反流性食管炎→Barrett食管→食管腺癌的发生、发展过程中,COL1A1mRNA的表达呈现渐进性增强,并与病期有关。结论 COL1A1基因的异常表达与反流性食管炎、Barrett食管和食管腺癌的发生、发展密切相关;COL1A1mRNA的表达水平可用作监测Barrett食管和食管腺癌早期发生及干预治疗效果的有用指标。     

大肠癌Rb和CyclinD1基因的表达及其临床意义

目的　探讨大肠癌中Rb和CyclinD1基因的表达水平及临床意义。方法　采用免疫组化SP法检测 4 0例大肠癌和 10例正常大肠组织中Rb和CyclinD1的表达。结果　 10例正常大肠黏膜中均可见Rb阳性表达 ,而大肠癌中Rb阳性率仅为 4 5 % ;CyclinD1在正常黏膜中几乎未见阳性表达 ,在癌组织中阳性率为 4 0 % ;Rb表达与大肠癌淋巴结转移相关 (P <0 .0 5 ) ,Rb阳性者的五年生存率显著高于Rb阴性者 (P <0 .0 5 ) ,而与肿瘤组织学分级和Dukes分期均无相关性 ;CyclinD1表达与大肠癌各临床病理参数无显著相关性。结论　Rb表达缺失和CyclinD1过度表达均与大肠癌的发生有关 ,可作为判断大肠癌预后的重要指标。     

THE CLINICAL SIGNIFICANCE OF MONITORING CARBOHYDRATE ANTIGEN CA50 IN COLORECTAL CARCINOMA

AntigenssuchasQ-fetoproteinandcarcinoembryonicantigenhavebeenusedasmarkersforthediagnosisandpostoperativeevaluationofpatientswithmalignanttumors.Lindholmetaldetectedamalignanttumorcelllinebyusinghybridomatechniquein1983,themonoclonalantibodiessecretedbythecelllineweredefinedasC50.TheantigenrecognizedbyC50monoclonalantibodywasnamedCA50.Itwasraisedagainstacolorectaladenocarcinomacellline.CA50antigenexistsonthecellsurfaceintheformoflipidorlipoproteincomplex,belongingtothemarkerofsphingoglycoli…     

尾型同源盒基因-2过表达对结肠癌细胞生物学性状的影响

目的观察尾型同源盒基因-2(CDX2)对结肠癌细胞生物学性状的影响,探讨CDX2基因在结肠癌发生发展中的作用。方法克隆人结肠CDX2基因,构建含CDX2的pEGFP-C1-CDX2表达载体,同时设pEGFP-C1空载体组及空白对照组。利用脂质体法将pEGFP-C1-CDX2和pEGFP-C1质粒分别转染Lovo细胞,应用Western blotting技术检测Lovo细胞中CDX2基因的表达;应用MTT法及流式细胞仪检测CDX2对Lovo细胞增殖、凋亡及周期的影响;应用ELISA法检测CDX2过表达对Lovo细胞基质金属蛋白酶-2(MMP-2)表达的影响。结果成功构建含CDX2真核表达载体。瞬时转染pEGFP-C1-CDX2的Lovo细胞48 h有较高水平的CDX2蛋白的表达;与pEGFP-C1空载体组及空白对照组比较,转染pEGFP-C1-CDX2组的Lovo细胞在转染72 h时生长未见明显抑制,G1期及S期所占比例无明显改变,凋亡率亦无明显增加(P均>0.05),但MMP-2分泌明显减少(P<0.05)。结论 CDX2过表达对Lovo细胞增殖、凋亡及周期无明显影响,但能够明显减弱Lovo细胞的侵袭力,从而对结肠癌浸润转移起抑制作用。     

血管内皮生长因子C及淋巴管密度的表达对判定Ⅱ期结直肠癌预后的意义

目的 分析56例Ⅱ期结直肠癌术后经病理证实的癌组织中血管内皮生长因子C(vascular endothelial growth factor-C, VEGF-C)表达和淋巴管密度(lymphatic vessel density, LVD)与其临床病理特征的关系及其对预后判定的意义,为Ⅱ期结直肠癌患者的综合治疗提供有意...