Galectin-3: differential expression between small-cell and non-small-cell lung cancer

AIMS: To compare the histological expression of galectin-3 in different lung cancers, including small-cell lung cancer (SCLC) and non-small-cell lung cancer (NSCLC). Lung cancer is the leading cause of cancer deaths in the UK. Galectin-3 is a beta-galactoside binding protein with a controversial role in malignant transformation. SCLC metastasizes early and is initially chemosensitive; NSCLC metastasizes later, offering the chance of surgical cure, but is much less chemosensitive. Mixed tumours present a diagnostic and therapeutic problem, with a poorer response to therapy. Insight into the cellular mechanisms that govern metastasis and chemoresistance will profoundly influence the future management of this disease. METHODS AND RESULTS: In this study the histological expression of galectin-3 was assessed in a panel of lung tumour specimens, using the indirect streptavidin-biotin method. A striking difference in galectin-3 expression was observed between tumours, with high expression in NSCLC (42/47 samples) and low expression in SCLC (negative in 13/18, weak in 5/18). CONCLUSION: This differential expression of galectin-3 between histological types of lung carcinoma suggests that galectin-3 may have an important influence on tumour cell adhesion, apoptosis and the response of tumours to chemotherapy.     

P53表达预测非小细胞肺癌化疗的耐药性及养肺解毒方的干预治疗

目的 探讨P53表达对预测非小细胞肺癌(NSCLC)化疗耐药性的意义及采用中药养肺解毒方对其的干预治疗.方法 选择Ⅲ、Ⅳ期NSCLC患者96例,经纤维支气管镜或者CT引导下95经皮细针肺活检获取标本,通过免疫组织化学、RT-PCR检测P53表达.依据其结果,分为P53阳性组(n=54)和P53阴性组(n=42).依据治疗方案的不同,每组再分为吉西他滨(健择,1250 mg/m2)+顺铂(70mg/m2)治疗组(T1组,P53阳性,27例;T2组,P53阴性,21例)和健择(1250mg/m2)+顺铂(70mg/m2)+养肺解毒方(随症加减,1剂/d,1次/d)治疗组(T3组,P53阳性,27例;T4组,P53阴性,21例).治疗周期为21 d,共4个周期.观察各组病例治疗后的有效率和中位生存期(MST).结果 阳性组P53 mRNA相对吸光度A值(57.65±4.78)明显高于阴性组(24.83±1.81)(P＜0.05).T2组治疗有效率42.9%(9/21)高于T1组29.6%(8/27),MST(9.55±0.54)个月也长于T1组(7.82±0.48)个月(均P＜0.05).T4组治疗有效率57.1%(12/21)高于T3组37.0%(10/27),MST(11.03±0.63)个月长于T3组的(8.78±0.51)个月(均P＜0.05).而且T3组的治疗有效率和MST明显优于T1组,T4组优于T2组(均P＜0.05).结论 P53蛋白和mRNA基因高表达可以预测非小细胞肺癌的耐药性.中药养肺解毒方可能通过降低非小细胞肺癌化疗耐药来提高疗效.     

CRKL在人非小细胞肺癌中的表达及意义

目的 CRKL(Crk-Like)基因在肺癌细胞系中存在着一定程度的扩增,我们将探讨接合物蛋白CRKL在非小细胞肺癌中的表达情况及与临床病理因素的关系。方法采用免疫组织化学和WB方法,分别检测了131例非小细胞肺癌组织和30例新鲜的非小细胞肺癌标本中CRKL的表达情况。结果 CRKL在正常支气管上皮细胞中呈阴性表达,而在肺癌组织中有44.3%(58/131)的病例存在阳性表达,WB检测发现肺癌中CRKL的表达明显高于对应的癌旁正常肺组织,CRKL蛋白在肺腺癌中的阳性表达为60.34%明显高于鳞癌(P=0.001),其表达与肺癌的低分化、高p-TNM分期(P=0.0035)、高Ki-67(P=0.0062)增殖指数和不良预后明显相关(P=0.0183)。结论 CRKL在肺癌中高表达,并与分化、分期和不良预后相关,提示CrkL蛋白可能在非小细胞肺癌的发生、发展中发挥作用。     

Increased expression of a Myc target gene Mina53 in human colon cancer

Mina53 is a novel Myc target gene that we previously demonstrated to be involved in cell proliferation. We studied, here, the expression of Mina53 in colon cancer to examine its possible role in carcinogenesis. We generated a specific monoclonal anti-human Mina53 antibody and found that colon tumor cell lines expressed Mina53 highly. We also found that expression of Mina53 was elevated in colon tumor tissues by immunoblotting analysis. Tissue sections of 23 surgical cases of adenocarcinoma and 1 case of adenoma were stained immunohistochemically, and the expression of Mina53 was found to be elevated in all of the adenocarcinomas compared to adjacent nonneoplastic tissues, which showed little staining. Deeply invading tumors as well as tumors that have invaded lymphatic vessels showed strong immunoreactivity against anti-Mina53 antibody. Mina53 was expressed in all pathological grades of cancer as well as in the adenoma. Staining patterns of Ki-67, a biomarker for cell proliferation, were similar to those of Mina53 in most cases, but the percentage of tumor cells stained by anti-Mina53 was higher. Although anti-Ki-67 antibody strongly stained some well-proliferating nonneoplastic cells including cells in the deeper part of the crypts and in lymphoid germinal centers, antibody to Mina53 rarely stained those cells. Suppression of mina53 expression severely suppressed proliferation of colon tumor cells in vitro. Together, our results indicate that the elevated expression of Mina53 is a characteristic feature in colon cancer, one that may have therapeutic applications.     

FGFR1 expression and gene copy numbers in human lung cancer

FGFR1 is a receptor tyrosine kinase of which the ligands belong to the fibroblast growth factor family. To evaluate the significance of FGFR1 in lung cancer, we analysed tumours by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Tissue microarrays were constructed containing 380 lung cancer samples including squamous cell carcinomas (SCC), adenocarcinomas (ADC), non-small cell lung cancer not otherwise specified, metastases, neuroendocrine tumours, large cell lung cancer and small cell lung cancer. FGFR1 expression was analysed by IHC and scored semi-quantitatively by a four-tier approach (0, 1, 2, 3). Using dual-colour interphase FISH with probes specific for the locus on 8p12 and the centromere of chromosome 8 (CEN8), copy numbers of FGFR1 were determined. High expression of FGFR1 was associated with increased FGFR1 gene copy numbers in squamous cell carcinoma (p?相似文献   

人肺癌相关抗原基因ALT04-AG表达抑制对人肺癌细胞株生长及基因表达谱的影响

目的探讨抑制人肺癌相关抗原基因ALT04-AG表达对细胞生长特性及相关基因表达的影响。方法重组反义ALT04-AGRNA真核表达质粒,经脂质体介导转染人肺癌细胞株(L78),以MTT、FCM法分析转染细胞生长特性,以Northern b lot、免疫组化染色及基因芯片技术检测相关基因的表达。结果构建了重组表达反义ALT04-AGRNA的真核表达质粒[pALT04-AG(as)],经该质粒转染及二氟甲基鸟氨酸(d ifluorom ethylorn ith ine,DFMO)处理的L78细胞均引起ALT04-AG表达下调及细胞的增殖抑制,后者还导致细胞凋亡比例增加。结论pALT04-AG(as)转染L78细胞或用DFMO抑制多胺生物合成,可通过对相关基因表达的调控促使L78细胞恶性表型逆转,而后者的作用更为广泛。本结果为探索肿瘤的诊断与治疗提供了有意义的线索。     

人肺癌组织中单羧酸转运泵基因mRNA表达的变化

本文应用RT－PCR方法扩增了人正常肺组织、肺癌组织中的MCT1、MCT2及β-actin基因mRNA,以人β-actin基因RT－PCR产物作为外参照,通过比较同一样本组中及样本组间MCT1、MCT2相对于β-actin的RT－PCR产物的光密度比值,来分析判断MCT1、MCT2mRNA表达的高低。结果显示,MCT1在人正常肺组织、肺癌组织中的mRNA表达均高于MCT2;MCT1、MCT2基因mRNA在人肺癌组织中的表达明显高于癌旁正常肺组织。推测MCT1可能在肿瘤组织细胞内的pH调节方面发挥重要作用。     

Smoking and cancer-related gene expression in bronchial epithelium and non-small-cell lung cancers

Tobacco smoking is the leading cause of lung cancer worldwide. Gene expression in surgically resected and microdissected samples of non-small-cell lung cancers (18 squamous cell carcinomas and nine adenocarcinomas), matched normal bronchial epithelium, and peripheral lung tissue from both smokers (n = 22) and non-smokers (n = 5) was studied using the Affymetrix U133A array. A subset of 15 differentially regulated genes was validated by real-time PCR or immunohistochemistry. Hierarchical cluster analysis clearly distinguished between benign and malignant tissue and between squamous cell carcinomas and adenocarcinomas. The bronchial epithelium and adenocarcinomas could be divided into the two subgroups of smokers and non-smokers. By comparison of the gene expression profiles in the bronchial epithelium of non-smokers, smokers, and matched cancer tissues, it was possible to identify a signature of 23 differentially expressed genes, which might reflect early cigarette smoke-induced and cancer-relevant molecular lesions in the central bronchial epithelium of smokers. Ten of these genes are involved in xenobiotic metabolism and redox stress (eg AKR1B10, AKR1C1, and MT1K). One gene is a tumour suppressor gene (HLF); two genes act as oncogenes (FGFR3 and LMO3); two genes are involved in matrix degradation (MMP12 and PTHLH); three genes are related to cell differentiation (SPRR1B, RTN1, and MUC7); and five genes have not been well characterized to date. By comparison of the tobacco-exposed peripheral alveolar lung tissue of smokers with non-smokers and with adenocarcinomas from smokers, it was possible to identify a signature of 27 other differentially expressed genes. These genes are involved in the metabolism of xenobiotics (eg GPX2 and FMO3) and may represent cigarette smoke-induced, cancer-related molecular targets that may be utilized to identify smokers with increased risk for lung cancer.     

The expression of beta-catenin in non-small-cell lung cancer: a clinicopathological study

AIMS: To investigate the expression of beta-catenin in non-small-cell lung cancer (NSCLC) and its clinical significance. METHODS: 101 patients were surgically treated for NSCLC by lobectomy or pneumectomy with systematic lymph node dissection. Follow up was available in all patients, ranging from 24 to 110 months. Immunostaining of tissue sections from primary tumours and (when present) their lymph node metastases was performed and evaluated using a monoclonal antibody against beta-catenin. Correlations were investigated between beta-catenin immunostaining in primary tumours and E-cadherin immunostaining (data available from a previous study), lymph node stage, and survival. RESULTS: There were significant correlations between scores for beta-catenin immunostaining and E-cadherin immunostaining in primary tumours (p = 0.007), and between the beta-catenin immunostaining score in primary tumours and in their lymph node metastases (p = 0.006). An inverse correlation was found between the beta-catenin immunostaining score in primary tumours and lymph node stage N0, N1, or N2 (p = 0.03). According to the Kaplan-Meier survival estimate, the level of beta-catenin expression in primary tumours was a statistically significant prognostic factor (p = 0.01). CONCLUSIONS: Reduced beta-catenin expression in surgically treated NSCLC is clearly associated with lymph node metastasis and an infavourable prognosis. The existence of a functional relation between E-cadherin and beta-catenin is supported by the results of this clinicopathological study.     

PGRN基因沉默对人非小细胞肺癌A549细胞增殖及迁移的影响

目的:探讨小干扰RNA(small interference RNA,si RNA)介导的颗粒蛋白前体(progranulin,PGRN)基因沉默对非小细胞肺癌A549细胞增殖、迁移和凋亡的影响及其机制。方法:分别用q PCR和Western blot法检测A549细胞和正常人支气管上皮(HBE)细胞中PGRN的m RNA和蛋白表达水平。采用脂质体转染法将PGRNsi RNA转染A549细胞,采用q PCR和Western blot法验证PGRN表达的变化;应用MTT实验检测细胞活力;活细胞计数法和结晶紫染色实验检测细胞增殖能力;划痕愈合实验和Transwell实验检测细胞迁移能力;并用Western blot法检测增殖细胞核抗原(PCNA)、细胞周期蛋白D1(cyclin D1)、Bcl-2和Bax的蛋白表达水平以及PGRN下游信号通路中细胞外信号调节激酶1/2(ERK1/2)和蛋白激酶B(Akt)的磷酸化水平。结果:PGRN在A549细胞中的m RNA和蛋白水平均明显高于HBE细胞(P0.05);转染PGRN-si RNA后A549细胞中PGRN的m RNA和蛋白水平均明显下调,细胞活力、增殖能力以及迁移能力均明显降低(P0.05)。沉默PGRN基因的表达,可下调PCNA、cyclin D1和Bcl-2的蛋白表达,而上调Bax的蛋白表达,且磷酸化的ERK1/2(p-ERK1/2)和磷酸化的Akt(p-Akt)的蛋白水平明显降低(P0.05)。结论:PGRN基因沉默能明显抑制非小细胞肺癌A549细胞的增殖和迁移能力,PI3K/Akt和MAPK/ERK信号通路可能在该过程中发挥重要作用。     

晚期非小细胞肺癌ERCC1基因表达量的临界值研究及验证

目的 检测200例非小细胞肺癌FFPE样本中ERCC1的相对表达量,确定判别ERCC1表达量等级的临界值并对其进行回顾性验证.方法 采用实时荧光定量PCR技术检测FFPE样本中ERCC1和内参基因的表达量,并通过2-ΔCt法计算ERCC1的相对表达量.以其中位值为判别ERCC1表达量等级的临界值,并通过患者应用铂类化疗药物的短期、长期疗效进行回顾性验证.结果 200例FFPE样本中,ERCC1和内参基因均可检出的检出率为89.0％.ERCC1相对表达量与患者年龄、性别、分型、分期及有无吸烟史等差异均无显著性(P＞0.05).高表达、低表达ERCC1患者在应用药物后的客观有效率分别为22.0％、53.7％ (P ＜0.05).采用COX模型进行多因素回归分析,发现ERCC1表达量是影响患者无进展生存、总体生存的独立因素(P＜0.05).ERCC1高表达、低表达患者接受铂类化疗药物治疗的中位无进展生存时间分别为8个月、14个月,差异有显著性(P=0.018);ERCC1高表达、低表达患者中位总体生存时间分别为10个月、15个月,差异有显著性(P =0.028).结论 判定非小细胞肺癌ERCC1表达量等级的临界值适合进行后续验证,为相关检测标准的制定提供依据.     

血小板因子4基因转染对肺癌细胞中血管生成因子基因表达的影响

目的：进一步探讨人血小板因子4（hPF4）抑制血管生成的作用机制,方法：构建含全长hPF4cDNA重组真核表达载体pcDNA3-hPF4,北朝鲜其转染到有肺巨细胞癌细胞系PLA801D细胞内,应用RT-PCR及免疫组化法观察肿瘤细胞自分泌的血管内皮生长因子（VEGF）、碱性成纤维细胞生长因子（bFGF)、白细胞介素8（IL-8）等的mRNA和蛋白表达水平的变化,结果：导入pcDNA3-hPF4,PLA801D细胞能稳定表达hPF4mRNA,其VEGF、bFGF、IL-8的mRNA和蛋白表达水平均明显降低,hPF4可直接调控VEGF、bFGF、IL-8等基因转录,影响其蛋白质生物合成,结论：提示hPF4可直接下调肿瘤细胞自分泌的VEGF、bFGF及IL-8等血管生成因子基因转录,抑制肿瘤细胞释放肿瘤血管生成因子,是hPF4抗血管生成抑制肿瘤细胞生长的机制之一。     

Prognostic significance of RIN1 gene expression in human non-small cell lung cancer

Ras interaction/interference 1 (RIN1), originally identified as a Ras effector protein, has been implicated in tumorigenesis and development of human cancers. The aim of this study was to detect RIN1 expression in human non-small cell lung cancer (NSCLC) and to analyze its association with prognosis of NSCLC patients. Quantitative real-time RT-PCR was performed to examine the expression of RIN1 mRNA in 25 cases of NSCLC and corresponding non-tumor tissue samples. Immunohistochemistry was performed to detect the expression of RIN1 in 90 NSCLC tissues. We found that the expression levels of RIN1 mRNA in NSCLC tissues were significantly higher than those in corresponding non-tumor tissues. High-level RIN1 expression was observed in 53.3% (48 of 90 cases), and correlated with poor tumor differentiation (P = 0.024), TNM stage (P = 0.032), and lymph node metastasis (P = 0.018). Patients with high expression levels of RIN1 showed lower overall survival rate than those with low expression levels (P = 0.033). Multivariate analysis showed that high RIN1 protein expression was an independent prognostic factor for NSCLC patients (P = 0.021). Our study suggests that over-expression of RIN1 may play an important role in the progression of NSCLC and RIN1 expression may offer a valuable marker for predicting the outcome of patients with NSCLC.     

Tumour fragment spheroids from human non-small-cell lung cancer maintained in organ culture

Biopsy material from 17 human non-small-cell lung carcinomas (NSCLC) was maintained in agar overlay culture as tumour fragment spheroids for 40 days. A practical procedure for the formation of spheroids and organ culture is described. The mechanically dissociated tumour specimens showed a variation in their ability to generate spheroids that was not related to the ploidy or the histological differentiation of the biopsies. Light microscopic observations revealed a heterogeneous spheroid population with a mixture of tumour cells and stromal elements. Most of the histological elements normally found in human NSCLC could be seen in the spheroids. The cellular components in the spheroids varied between highly cellular to sparsely cellular, dominated by stromal elements. The squamous carcinomas were in general found to generate highly cellular spheroids more often than the adenocarcinomas. Spheroids with a different cellular content could be selected in vitro by using a morphometric technique. Diameter measurements showed a large variability in spheroid growth. Most of the spheroids decreased in size although bromodeoxyuridine labelling indicated active cell proliferation in the specimens. Frequent changes of medium did not affect spheroid growth. The culture system presented provides a model for studying the cellular heterogeneity as well as the biological characteristics of tumour tissue from individual patients in vitro.