BRI基因的表达与非小细胞肺癌发生及转移潜能的关系

目的 探讨BRI基因在人非小细胞肺癌中的差异表达情况及其与肺癌发生及转移能力形成的相关性。方法 应用逆转录-聚合酶链反应(RT-PCR)、Northern印迹杂交方法分析BRI基因在一对遗传背景相同、转移能力不同的人肺腺癌细胞系AGZY83-a和Anip-073中的差异表达,并进一步检测BRI基因在另外6个非小细胞肺癌细胞系(SPC-A-1,A549,95D,TKB-18,GLC-82,PAa)及30例临床肺癌标本中的表达情况。结果 BRI基因在2细胞系中存在明显的差异表达,在具有高转移潜能的Anip973中高表达。在其他6个细胞系中BRI的表达与肺癌转移潜能可能有关。同时发现与同一病例的正常肺组织相比,BRI基因在肺癌组织中高表达,其过表达在有淋巴结转移的肺癌为6／8,无转移者为45．5％(10／22),并且BRI基因在肺癌与正常肺组织之间存在转录本的差异。结论 BRI基因在肺癌中表达上调,其1．6kb转录本表达的增加可能与肺癌的发生和转移能力的形成有关。     

A murine monoclonal antibody raised against human non-small cell carcinoma of the lung

Murine monoclonal antibodies (MAbs) reactive with human non-small cell carcinoma of the lung (NSCCL) were produced following immunization with a membrane preparation of Calu-3, a human adenocarcinoma of the lung cell line grown in nude mice. Positive hybrids unreactive with normal liver membranes and human peripheral white blood cells were selected for further testing. One MAb (RS5-4H6) recognized an antigen expressed in a variety of lung and other carcinoma cell lines as detected by flow cytometry. Immunohistology showed a selectivity for normal and neoplastic lung epithelium, as well as other cancers. The antigen was detected by immunohistology in 87% of tumor specimens derived from the lung, breast, colon, kidney, and ovary. Most other normal human tissues were not stained but occasional cells of the stomach, salivary and other glands, as well as kidney tubules were reactive. This MAb is an IgG1. Western blot analysis indicated that the antibody reacts selectively with an antigen greater than 300 kD. The antigen is resistant to neuraminidase treatment and periodate oxidation, but sensitive to pronase treatment, suggesting that the epitope is peptide in nature. This antibody may be potentially useful as a targeting agent for radioimmunodetection and immunoconjugate therapy.     

Microvessel count predicts metastatic disease and survival in non-small cell lung cancer

The growth of newly formed vessels, or neoangiogenesis, represents an important step in both physiological and pathological situations: in particular, tumour growth and metastasis require angiogenesis. Microvessel count (MC), which represents a measure of tumour angiogenesis, has been associated with metastatic spread in cutaneous, mammary, prostatic, head and neck, and early-stage lung cancer. In this study, the role of tumour angiogenesis as a prognostic indicator was examined in 253 primary non-small cell lung cancer (NSCLC) patients. Microvessels were counted by highlighting endothelial cells with anti-Factor VIII monoclonal antibody (MAb) in methacarn-fixed tumour samples. In univariat analysis, MC (P<0·000001), sex (P=0·0036), histotype (P<0·014), tumour status (P<0·007), and vessel invasion (P<0·019) were significantly related to hilar and/or mediastinal nodal involvement. However, in the stepwise logistic regression analysis, MC (P<0·000003) retained the most important influence on nodal metastasis. The overall survival analysis calculated by the Kaplan-Meier method revealed that tumours with high MC (>25 vessels/field) were significantly associated with increased death risk (log-rank test P=0·00067; Cox's test P=0·00046; Gehan's Wilcoxon test P=0·00108). In 94 patients, the development of metastatic disease during follow-up was significantly related to MC. Indeed, patients who developed metastasis during follow-up showed a higher MC, either as a dichotomous (P=0·01) or as a continuous (P=0·003) variable, than patients who had developed no metastasis at the time of the analysis. Moreover, in the stepwise logistic regression analysis, MC retained the most important influence on distant metastases. Microvessel count, as a method for the quantitation of tumour angiogenesis, has an important prognostic role in non-small cell lung carcinomas.     

Selective p38 activation in human non-small cell lung cancer

The mitogen-activated protein kinase (MAPK) pathways transmit signals from the cell membrane to the nucleus. Activation of MAPK cascades may play a role in malignant transformation. We hypothesized that enhanced expression of one or more of these pathways would occur in human lung cancers. Using Western blot analysis of tissue homogenates from resected non- small cell lung cancers and matched non-neoplastic lung tissue, we determined that only activated p38 was consistently increased in tumor compared with normal tissue. In vitro kinase assays confirmed that the levels of activated MAPK correlated with the activity of the enzymes, and immunohistochemical analysis confirmed the cellular localization of the activated MAPKs. We incubated a lung cancer cell line in a hypoxic chamber to simulate the hypoxic environment in solid lung tumors, but found no increase in p38 activation. Contrary to our expectations, ERK and JNK, the MAPK pathways traditionally associated with cell growth and perhaps malignant transformation, were not consistently activated in the human lung tumor samples. However, p38, a MAPK usually associated with stress responses, growth arrest, and apoptosis, was activated in all of the human lung cancer samples, suggesting an additional role for this pathway in malignant cell growth or transformation.     

人非小细胞肺癌组织中自发性癌细胞凋亡的研究

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p73：人类非小细胞肺癌细胞株中的肿瘤抑制基因(英文)

目的：p73基因是与p53相类似的基因,在人类神经母细胞瘤中被假定作为肿瘤抑制基因。为了证实p73在非小细胞肺癌中是否也是肿瘤抑制基因，我们应用StyⅠ内切酶多态分析法研究了6株非小细胞肺癌细胞的等位基因表达模式。方法：利用RT-PCR检测p73基因在这6株肺癌细胞中转录水平的表达，同时用免疫组织化学方法检测5株非小细胞肺癌细胞所诱发的裸鼠种植瘤中P73蛋白的表达。结果：p73基因在这6株非小细胞肺癌中均为纯合性等位基因表达，而GC/GC基因型为主要类型。p73基因在转录水平和蛋白水平完全丧失表达。结论：根据实验结果可推测，p73在这6株非小细胞肺癌细胞中仍扮演了肿瘤抑制基因的角色。     

人非小细胞肺癌细胞中端粒酶活性的检测与研究

目的 研究非小细胞肺癌细胞癌组织中端粒酶活性表达,探讨端粒酶生表达与非小细胞肺癌发生发展的关系。方法 收集经手术及病理证实的非小细胞肺癌４８例标本,１２例肺癌癌旁肺组织、７例非肿瘤病例所取正常肺组织作对照。用端粒酶检测试剂盒检测组织本端粒酶活性。结果 ７５．００％（３６／４８）非小细胞肺癌组织标本检出端粒酶活性,８．３３％（１／１２）癌旁肺组织检出端粒酶活笥,７例非肿瘤标本所取的正常肺组织均未检出     

Molecular pathology of non-small cell lung cancer

Our increasing understanding of the molecular pathogenesis of non-small cell lung cancer (NSCLC), particularly adenocarcinomas, has opened the door to ‘personalised medicine’ and the advent of new therapeutic strategies. Over the last few years new drugs, or classes of drugs, have been licensed and entered clinical practice for use in advanced NSCLC. The activity of these drugs is dependent on the presence of specific molecular or protein changes in cancer cells which are usually identified using ‘companion diagnostic tests’ specifically designed for this purpose. Pathologists and Pathology Departments have had to forge new links with Clinical Scientists in order to facilitate these additional investigations on the, often limited, tissue obtained for diagnosis. This collaboration plays a critical role in providing the link that allows integration of the traditional morphological diagnosis with the results of these new ‘companion diagnostic’ tests to guide patient management.     

hnRNP A2/B1 mRNA在非小细胞肺癌和转移淋巴结中表达及意义

目的：探讨肺癌早期诊断基因核内不均一核糖体表达对肺癌淋巴结微转移的作用。观察hnRNP A2/B1 在各组织类型肺癌中的表达。方法：42例非小细胞肺癌患者开胸手术纵隔淋巴结清扫获取原发灶及纵隔淋巴结病理标本,12例肺良性病变手术切除标本作正常对照。荧光定量PCR检测切除肿块和淋巴结的hnRNP A2/B1 mRNA表达。结果：(1)转移性淋巴结中hnRNP A2/B1 mRNA 表达水平561.393显著高于无转移淋巴结156.669 (P＜0.05)。(2)Ⅲ期肺癌组织中hnRNP A2/B1 mRNA 表达水平305（249,533）显著高于Ⅰ期和Ⅱ期肺癌组织(2＝13.453,P＜0.01)。(3)hnRNP A2/B1 mRNA在鳞癌和腺癌中的表达水平分别为207(152,305)和249(177,476), Z=-0.899,P>0.05。(4)58组淋巴结阳性,84组阴性,42个病人142组区域淋巴结hnRNP A2/B1 mRNA阳性率为40.84%(58/142),12例肺良性疾病组织均阴性,HE染色42个病人的50组转移淋巴结阳性率为35.21%,与FQ-PCR 58枚（52.11%）无显著差异（P>0.05）。结论：NSCLC 癌组织存在着hnRNP A2/B1 的过度表达。NSCLC癌组织中hnRNP A2/B1表达水平与淋巴结转移状态及P-TNM 分期有关。 hnRNP A2/B1 检验敏感性、特异性高,可用于肺癌的辅助诊断的标记。     

Carbonic anhydrase-related protein VIII increases invasiveness of non-small cell lung adenocarcinoma

Carbonic anhydrase-related protein VIII (CA-RP VIII) is believed to be an oncofetal antigen and is overexpressed in colorectal and non-small cell lung cancer. However, the pathobiological properties of CA-RP VIII in lung cancer remain unclear. In the present study, we examined ultrastructural changes caused by exogenous CA-RP VIII expression in a well-differentiated lung adenocarcinoma cell line, PC-9. Many vacuoles lined by cilia, sometimes large vacuoles pushing the nuclei to one side, were found in the cytoplasm of CA-RP VIII-expressing PC-9 cells, but not in control PC-9 cells. Moreover, signet-ring cells containing abundant intracytoplasmic mucin were often found among CA-RP VIII-expressing PC-9 cells, but rarely among control PC-9 cells. We subsequently examined CA-RP VIII expression in atypical adenomatous hyperplasia and early-stage lung adenocarcinoma (Stage Ia). Significant expression of CA-RP VIII was observed in invasive lung adenocarcinoma but not in noninvasive adenocarcinoma. Interestingly, CA-RP VIII was strongly expressed in signet-ring cell cancer and invasive mucinous adenocarcinoma components. CA-RP VIII also appeared to enhance the invasiveness of PC-9 cells in Matrigel invasion assay. The present findings suggest that CA-RP VIII expression in lung adenocarcinoma is related to cancer cell invasion.     

非小细胞肺癌新辅助免疫治疗的研究进展

肺癌的发病率和死亡率普遍较高,严重威胁大众身体健康。近年来,免疫检查点抑制剂的出现给晚期非小细胞肺癌的治疗带来了新的希望。结合理论基础,科学家推测非小细胞肺癌患者可能受益于新辅助免疫治疗。随着研究的深入, 发现新辅助免疫治疗非小细胞肺癌具有较高的安全性和可行性,并可获得振奋人心的病理缓解率。但目前仍存在缺乏远期预后,以及尚未明确潜在受益人群、合理的治疗方案和疗程等问题。本文对新辅助免疫治疗非小细胞肺癌的发展过程、作用机制、研究现状以及与治疗相关的挑战进行综述。     

Simultaneous detection of genetic and immunological markers in non-small cell lung cancer: prediction of metastatic potential of tumor

The restriction fragment length polymorphism of c-Ha-ras-1 and L-myc genes and expression of cell surface effector molecules were studied to determine their potential utility as markers for assessing risk of metastasis in 84 lung cancer patients. We performed a comparative study of primary lung carcinomas, metastases, adjacent tissues and blood samples in a group of patients with lung cancer of different histological types, grade of differentiation and presence of regional and distant metastasis. No differences in the frequency of c-Ha-ras-1 rare alleles were found between lung cancer patients and unaffected controls. The detection of common a4-allele seems to be associated with metastasis and low differentiation of lung carcinomas. S-allele of L-myc was observed in 82.6% of patients with metastatic lesions. Homozygosity of L-allele patients was not evidence for distant metastasis and only 17.4% of these patients have metastatic lesions of the lymph nodes. The expression of HLA class I and receptor of transferrin (TrRec) were tested immunohistochemically in the same patients. In the group of squamous cell carcinomas with regional metastases the expression of HLA class I antigens was decreased [7/21 (33.3%) positive staining tumors versus 13/20 (65.0%) in the group without metastases]. The opposite situation was observed for TrRec. The data of restriction fragment length polymorphism of oncogenes and expression of two cell surface effector molecules, identified in the same patients, were combined. The registration of more than one poor marker, tested in individuals with squamous cell carcinoma, closely correlated with dissemination and advanced stage of the disease. Nearly 90% (20/22) of patients with well and moderately differentiated tumor revealed metastatic lesions versus 6.6% (1/15) of patients with manifestation of a single poor marker. Finally, proposals could be made for the development of a risk group that incorporates both clinical and molecular biology features in the prediction of metastasis.     

重组变构人TRAIL对非小细胞肺癌细胞的体外抑制作用

目的 研究重组变构人TRAIL(rmhTRAIL)对人非小细胞肺癌(NSCLC)细胞和人正常细胞的生长、抑制作用及其与化疗药的联合作用。方法 采用四唑盐(MTT)和集落形成试验法检测细胞的生长,并观察了与顺铂(DDP)联合应用对细胞生长的作用。结果 rmhTRAIL对多种。NSCLC细胞有显著的抑制作用,而对人正常细胞无影响;rmhTILAIL与DDP联合应用具有协同或增强对NSCLC细胞的抑制作用,而且对正常人细胞不表现毒性。结论 rmhTRAIL是一种安全、有效的抗肿瘤生物制品。     

The prevalence of human papillomavirus infection in Korean non-small cell lung cancer patients

Human papillomavirus (HPV) infection is a co-carcinogen of lung cancer and contributes to its pathogenesis. To evaluate the prevalence of HPV infection, polymerase chain reaction (PCR) was employed to detect HPV 16, 18, and 33 DNA in tumor tissues of 112 patients with non-small cell lung cancer (NSCLC) who underwent curative surgery from Jan. 1995 to Dec. 1998 at Severance Hospital, Seoul, Korea. The patients consisted of 90 men and 22 women. Nineteen patients were under 50 years old (17%), and 92 patients (82%) were smokers. Fifty-three patients had adenocarcinomas, while 59 patients had non-adenocarcinomas. Early stage (I and II) cancer was found in 64 patients (57.1%) and advanced stage (III and IV) found in 48 (42.9%). The prevalence of HPV 16, 18, and 33 were 12 (10.7%), 11 (9.8%), and 37 (33.0%), respectively. Smoking status, sex, and histologic type were not statistically different in the presence of HPV DNA. The presence of HPV 16 was more common in younger patients and HPV 18 was more common in advanced stage patients. This study showed that the prevalence rate of HPV 16 and 18 infections in NSCLC tissue was low, suggesting HPV 16 and 18 infections played a limited role in lung carcinogenesis of Koreans. However, the higher prevalence of HPV 33 infections in Korean lung cancer patients compared to other Asian and Western countries may be important and warrants further investigation.     

Endoglin在非小细胞肺癌中的表达及临床意义

目的 研究不同转移潜能非小细胞肺癌(NSCLC)细胞系中以及癌与癌旁组织中endoglin (ENG)的表达情况,探讨ENG在NSCLC的发生发展和转移复发中的作用.方法 体外培养5株NSCLC细胞及1株正常支气管上皮细胞(HBE),随机选取22例NSCLC手术切除后癌与癌旁组织样本,分别采用real time PCR及Western blot法检测其中ENG mRNA和蛋白表达情况.采用x2检验分析ENG的表达和临床资料之间的相关性.结果 高转移性3株肺癌细胞系中ENG mRNA和蛋白表达显著升高,而正常细胞株及低转移性2株肺癌细胞中ENG表达缺失.22例肺癌样本中,其中19例癌组织中ENG mRNA和蛋白呈阳性表达(86.36％),且表达量明显高于相对应的癌旁组织(P＜0.01).ENG表达与患者临床病理特征年龄、性别、肿瘤大小、临床分期、病理分级、组织类型等均无关,而与患者淋巴结转移情况密切相关(P＜0.01).结论 ENG在NSCLC中的表达与淋巴结转移显著相关,有可能作为预测NSCLC转移及预后的分子标志.     

川陈皮素对非小细胞肺癌A549细胞的抑制作用

目的:观察川陈皮素(5,6,7,8,3′4′-hexamethoxyflavone)对非小细胞肺癌A549细胞的抑制作用并研究其作用机制。方法:以不同浓度的川陈皮素作用于A549细胞,分别用MTT法、细胞生长曲线研究川陈皮素对A549的增殖抑制作用,Giemsa染色观察细胞形态变化。结果:MTT提示川陈皮素浓度为4mg·L-1~128mg·L-1时,对A549的48、72小时抑制率为15%~85%,48小时IC50为25.7mg·L-1,72小时IC50为16.7mg·L-1;生长曲线提示川陈皮素对A549细胞的抑制作用呈明显时效和量效关系;Giemsa染色后细胞出现明显的凋亡形态学变化。结论:川陈皮素对非小细胞肺癌A549细胞具有增殖抑制作用,其机制可能是诱导细胞凋亡。     

非小细胞肺癌患者组织原发灶和转移灶中ROS1融合基因的研究

目的：研究晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)原发灶和转移灶ROS1融合基因阳性率,探讨其相关性。方法：收集2013年1月至2015年5月中国人民解放军北京军区总医院、军事医学科学院附属医院、浙江省肿瘤医院、大连大学附属中山医院和山西医学科学院山西大医院原发灶384例,其中配对转移灶246例,统计得出ROS1融合基因阳性率并分析原发灶与转移灶ROS1融合基因的一致性、ROS1融合基因阳性与临床基线资料间的关系。结果：ROS1融合基因阳性率原发灶为2.60%(10/384)。ROS1融合基因阳性率配对原发灶为2.85%(7/246),配对转移灶为1.63%(4/246),配对的246对原发灶、转移灶组织中,转移灶融合基因阳性而对应的原发灶融合基因阴性1例,原发灶融合基因阳性而对应的转移灶融合基因阴性4例,转移灶较原发灶检出ROS1融合基因阳性率高,两者差别有统计学意义(χ2=52.341,P=0.000);转移灶和原发灶ROS1融合基因阳性的一致率好(κ=0.536,P=0.000),通过转移灶判断原发灶融合阳性的情况,敏感性为42.86%(3/7),特异性为99.58%(238/239)。结论：非小细胞肺癌中转移灶可以预测原发灶ROS1融合基因情况,在难以取得原发灶的情况下转移灶可以作为ROS1融合基因测的备选手段。     

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.