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。     

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。     

nm23 mRNA、CD44s和CD44v6在胃腺癌组织中的表达及其意义

目的　探讨nm2 3mRNA、CD4 4s和CD4 4v6在胃腺癌组织中的表达及其与胃腺癌病理学分级、浸润程度和转移之间的相关性。方法　应用免疫组织化学和mRNA原位杂交方法检测了 6 2例胃组织标本中nm2 3mRNA、CD4 4s和CD4 4v6的表达情况。结果　nm2 3mRNA在胃正常组织和癌组织中的表达无显著性差异 (P >0 .0 5 ) ,且与胃腺癌浸润和病理学分级无相关性 (P >0 .0 5 ) ,但与胃腺癌淋巴结转移呈明显负相关 (r =- 4 .93)。CD4 4s在胃腺癌与胃正常组织中的阳性表达率分别为 4 8% (19 4 0 )和 13.6 % (3 2 2 ) ,有显著性差异 (P <0 .0 1)。CD4 4v6只在胃腺癌中表达 ,阳性率为 6 3.3% (2 5 4 0 ) ,同时CD4 4v6表达与胃腺癌分化程度、淋巴结转移及其浸润程度呈明显正相关 (r =5 .0 4 )。结论　CD4 4v6高表达和nm2 3mRNA低表达患者淋巴结转移率高。联合检测CD4 4v6和nm2 3mRNA能更准确地判断胃腺癌的转移状态。CD4 4v6过量表达与胃腺癌的分化程度及浸润相关。     

人肺腺癌细胞系A549细胞双向电泳-飞行时间质谱研究

目的初步分析人肺腺癌细胞系A549细胞蛋白质表达情况,从蛋白组水平探讨肺腺癌发病的分子机制。方法应用2-DE技术对人肺腺癌细胞系A549细胞总蛋白进行分离,获得蛋白质表达谱;应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)结合生物信息学进行蛋白质鉴定。结果3块胶平均蛋白斑点数为1 138±49,平均匹配点数为986±32,匹配率为85.8%。随机选取背景清晰、分辨清楚、蛋白含量较高的20个斑点进行MALDI-TOF-MS分析,共获得了18个肽质量指纹图谱(PMF)。将PMFs质量数据通过Aldente软件查询SWISS-PROT数据库,根据匹配片段及氨基酸序列覆盖率等,初步鉴定出15种蛋白质。根据功能,这些蛋白质可分为:①基本代谢相关的酶类:果糖2-磷酸醛缩酶、视网醛脱氢酶1(RALDH1)、吡多醛激酶;②细胞骨架类:蛋白细胞角蛋白8(CK8)、-β2链微管蛋白;③应激相关蛋白:蛋白二硫化物异构酶A3(PDIA3);④信号转导分子:膜联蛋白1(ANX1)、膜联蛋白4(ANX4);⑤分子伴侣:热休克蛋白60(HSP60)、热休克蛋白-β1、抑制素(PHB);⑥转录及翻译相关蛋白:不均一性核糖核蛋白H(hnRNP H);⑦杂类:Rgr protein、NPM、PCBP1。结论应用2-DE/MALDI-MS方法获得了较为理想的人肺腺癌细胞系A549细胞2-DE蛋白质表达谱,初步鉴定了15种蛋白质。     

Identification of serum biomarkers for diagnosing stage I lung adenocarcinoma by MALDI-TOF mass spectrometry

Objectire To identify specific biomarkers that could improve early diagnsis of lung adenuearcinoma using matrix-assisted laser desorptian/ionization (MALDI) technology. Methods Serum samples were isolated from 17 patients with stage I lung adenuearcinoma and 17 age- and sex-matched healthy controls, and the serum proteomic profiles were obtained by matrix-assistcd laser desorption ionization time of flight (MALDI-TOF) mass spectrometry. Results Compared with healthy control group, two highly expressed potential biomarkers were identified with the relative molecular weights of 6 631.64 Da and 4 964. 21 Da. The two best novel protein peaks were automatically chosen for the system training and the development of the constructed model. The constructed model was then used to test an independent set of masked serum samples from 15 lung adenocarcinoma patients and 22 healthy individuals. The analysis yielded a sensitivity of 93.3 %, and a specificity of 95.5 %. Conclusion These results suggest that MALDI-TOF-MS ProteinChip technology is a quick, convenient, and high-output analyzing method that is capable of selecting several relatively potential biomarkers from the serum of lung adenocarcinoma patients and may have a clinical value in the future, and will provide clues to identifying new serologic btomarkers of lung adenocarcinoma.     

P27~(Kipl)与Ki-67抗原在结直肠腺瘤、腺癌组织中的表达及意义

目的　通过检测P2 7Kipl及Ki 6 7抗原在结 (直 )肠腺瘤、腺癌中的表达 ,探讨其与临床病理指标及预后的关系。方法　采用免疫组织化学方法检测了 2 0例正常结 (直 )肠粘膜、16例腺瘤、6 2例腺癌的P2 7Kipl及Ki 6 7抗原的表达。结果　正常粘膜、腺瘤至腺癌的等级与P2 7Kipl表达等级呈负相关 ,而与Ki 6 7抗原表达等级呈正相关。腺瘤与腺癌细胞质P2 7Kipl表达有显著性差异 (P <0 .0 5 )。腺癌组P2 7Kipl表达与Ki 6 7抗原表达、肿瘤有无转移、Dukes’分期及国际TNM分期呈负相关 ;Ki 6 7抗原表达与有无转移呈正相关。P2 7Kipl表达阳性腺癌患者无病生存时间长于表达阴性者。结论　在结 (直 )肠肿瘤的发生、发展及转移过程中 ,P2 7Kipl表达下调 ,Ki 6 7抗原表达上调 ;P2 7Kipl表达对评估结 (直 )肠癌患者预后有一定帮助。     

益气养血保元方对肺癌细胞PCNA、p53蛋白表达的影响

采用免疫组化法观察益气养血保元胶囊 (OQP)对体外培养人肺癌 A- 549细胞 PCNA、p53蛋白异常表达的影响。结果 :OQP作用后 A- 549细胞 PCNA、p53蛋白表达减弱。提示 :OQP可抑制 PC-NA、p53蛋白表达 ,其抑癌作用机理可能与抑制 PCNA、p53蛋白表达有关。     

E-钙黏附素及增殖细胞核抗原在胰腺癌组织中的表达及其意义

目的研究E-cadherin、增殖细胞核抗原(PCNA)在胰腺癌组织中的表达及临床意义。方法应用免疫组化方法对59例胰腺癌组织的E-cadherin、PCNA进行检测。结果E-cadherin在胰腺癌组织比相应的癌旁正常胰腺组织中表达明显减低,高表达率仅为13.6%(8/59),且各组间(与癌组织分化程度、淋巴结转移以及临床分期)E-cadherin表达差异有统计学意义(P<0.05);PCNA在胰腺癌组织比相应的癌旁正常胰腺组织表达明显增高,强阳性表达率为67.8%(40/59),且各组间PCNA表达差异有统计学意义(P<0.05);在胰腺癌组织中,E-cadherin与PCNA表达呈负相关(P<0.05)。结论E-cadherin与PCNA异常表达与胰腺癌的分化、增殖、侵袭以及转移有密切的关系。