ALK免疫组化切片重复利用进行FISH检测的可行性

目前肺癌是全球发病率和病死率增长较快的恶性肿瘤之一,其中非小细胞肺癌(non-small cell lung cancer,NSCLC)占80％以上,约5％的NSCLC出现间变性淋巴瘤激酶(anaplastic lymphoma kinase,ALK)重排[1].近年来克唑替尼等ALK基因靶向药物临床发展迅速,对晚期N...     

肺鳞状细胞癌EGFR/ALK的基因状态

目的:探讨肺鳞状细胞癌各亚型中EGFR和ALK的基因状态.方法:应用ARMS方法检测肺鳞状细胞癌石蜡组织中EGFR基因突变和ALK融合基因情况.结果:218例肺鳞状细胞癌样本中,E GFR基因突变率为4.59％(10/218),19del和L858R各为2.29％(5/218).ALK融合基因阳性率为6.14％(7/114).结论:肺鳞状细胞癌存在一定比例EGFR基因突变和ALK融合基因阳性,肺鳞状细胞癌EGFR基因和ALK融合基因常规检测不可忽视.     

食管鳞状细胞癌中CK7蛋白表达及预后意义

目的 观察细胞角蛋白7(CK7)在食管鳞状细胞癌中的表达及其与预后的关系.方法 采用免疫组化ELPS法检测512例食管鳞状细胞癌中CK7的表达,分析其表达与临床病理特征及患者预后的关系.结果 512例食管鳞状细胞癌中CK7蛋白阳性率为24.6％,CK7表达与肿瘤分化差、淋巴结转移和临床分期较晚相关(P＜0.05).生存...     

新疆地区口腔鳞状细胞癌中PLCE1蛋白的表达及临床意义

目的探讨PLCE1蛋白在新疆地区口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)中的表达及预后价值。方法采用免疫组化EnVision两步法检测124例OSCC及63例癌旁正常组织中PLCE1蛋白的表达,分析其表达与OSCC临床病理特征及预后的关系。结果OSCC及癌旁组织中PLCE1蛋白高表达率分别为41.9%(52/124)和0(0/63),差异有统计学意义(P<0.0001)。PLCE1诊断OSCC的ROC曲线下面积(AUC)为0.954(敏感度83.9%,特异性95.2%)。Kaplan-Meier及Cox单因素分析显示,PLCE1高表达(P=0.004、P=0.006)和T分期(P=0.002、P=0.004)是影响患者不良预后的危险因素。Cox多因素分析结果显示,PLCE1高表达(P=0.008)和T分期(P=0.005)是患者预后的独立因素。结论PLCE1蛋白在新疆地区OSCC组织中高表达,且与患者不良预后相关,可作为肿瘤患者不良预后的新型生物标志物。     

肺鳞状细胞癌中死亡相关蛋白激酶的表达

目的　检测肺鳞状细胞癌中死亡相关蛋白激酶 (DAP K)mRNA表达及细胞凋亡 ,探讨DAP K与细胞凋亡的关系及其在肺鳞状细胞癌发生、发展中的作用。方法　用原位分子杂交法检测 6 0例肺鳞状细胞癌、9例癌旁肺组织DAP KmRNA表达 ;用原位末端标记TUNEL法检测相应组织中细胞凋亡 ,计算凋亡指数 (AI)。结果　肺鳞状细胞癌的DAP KmRNA阳性表达率为 4 6 7% ,癌旁肺组织为 6 7 7% ,其阳性率高于肿瘤组织 (P <0 0 1 )。在肺鳞状细胞癌中 ,高分化癌DAP KmRNA阳性率为 70 % ,低分化癌为 2 3 3% ,高分化癌的DAP KmRNA阳性率高于低分化癌 (P <0 0 1 )。肺鳞状细胞癌的细胞AI为(0 6 72 8± 0 4 2 6 1 ) % ,癌旁肺组织中支气管肺泡上皮细胞AI为 (1 0 2 89± 0 2 4 33) % ,癌旁肺组织的AI高于肿瘤组织 (P<0 0 1 )。在肺鳞状细胞癌中 ,高分化癌的AI为 (0 5 82 3± 0 1 92 2 ) % ,低分化癌为 (0 4 4 6 0± 0 1 92 5 ) % ,高分化癌的AI高于低分化癌 (P <0 0 1 )。DAP KmRNA呈阳性表达的肺癌 ,其AI为 (0 5 31 7± 0 2 0 97) % ;DAP KmRNA呈阴性者 ,其AI为 (0 4 872± 0 1 91 8) % ,两组间差异有显著性 (P <0 0 5 )。在连续切片上 ,DAP KmRNA阳性细胞的分布区域与凋亡阳性细胞的分布相似。DAP KmRNA呈阳性表达     

肺鳞状细胞癌中p53蛋白表达及p53基因突变的检测

目的 通过检测肺鳞状细胞癌及癌旁组织中Ｐ５３蛋白积聚及相同癌组织中Ｐ５３基因的突变，探讨Ｐ５３蛋白积聚及Ｐ５３基因突变在肺鳞癌发病中的意义。方法 采用免疫组化和银染－ＰＣＲ－ＳＳＣＰ方法检测１２０例肺鳞癌及癌旁肺组织中Ｐ５３蛋白的状况及相同鳞癌组织中，Ｐ５３基因５、６、７、８外，显子突变的情况。结果 Ｐ５３蛋白了性率为５２．５％，Ｐ５３基因突变率为５６．７％，突变便数在第５、６、７、８外显子的分布     

肺鳞状细胞癌和腺癌中人乳头瘤病毒感染及p53蛋白表达的研究

肺癌是人类最常见的恶性肿瘤之一，严重威胁人类健康。肺癌主要与吸烟有关，其它因素如病毒感染与肺癌的关系也是目前研究的热点，其中人乳头瘤病毒(human papillary virus，HPV)感染与肺癌的关系引起了广泛注意。HPV是小型DNA肿瘤病毒，其治病主要是通过两种病毒基因蛋白影响细胞周期调节。约10％的肿瘤发现有HPV感染，主要见     

子宫颈鳞状细胞癌中PDHA1的表达及临床意义

目的 探讨丙酮酸脱氢酶E1亚单位α(pyruvate dehydrogenase E1 component subunit alpha,PDHE1α)(又称PDHA1)在子宫颈鳞状细胞癌(cervical squamous cell carcinoma,CSCC)中的表达及临床意义.方法 采用免疫组化EnVision法...     

ALK gene rearranged lung adenocarcinomas: molecular genetics and morphology in cohort of patients from North India

ALK gene rearrangement in the lung adenocarcinomas is the second most common (1.6–11.7% of NSCLC) targetable genomic change after EGFR mutations. However, the prevalence and clinicopathological features of ALK‐rearranged lung adenocarcinomas from North India are lacking. A total of 240 cases of lung adenocarcinoma were screened for EGFR mutations and for ALK expression. Smoking status, TNM stage, and treatment response were recorded in all cases. Out of 240 cases screened, 37 cases were positive for EGFR mutations and 17 cases (7.08%) showed ALK positivity with immunohistochemistry and break‐apart FISH. On excluding 37 EGFR mutation–positive cases, the incidence of ALK‐positive adenocarcinoma appears to be higher (17/203 cases, 8.03%). Eight were men and nine were women with mean age of 51.7 years. Majority (62.5%) were non‐smokers and had unresectable disease (70.6% stage IV, 17.6% IIIB). The morphological patterns noted were solid (12 cases), papillary (four cases), and micropapillary (one case). Signet ring (two cases) and clear cell change (one cases) were noted. Out of five patients who received crizotinib, three had partial response and two had stable disease. ALK‐rearranged lung adenocarcinomas account for a minor proportion of NSCLC with prevalence similar to that reported in literature. However, as contrast to published data in our series, patients were in older age group and had solid and papillary pattern on morphology with an aggressive course.     

荧光原位杂交技术检测肺癌患者痰液中部分染色体数量的改变

目的应用染色体特异性探针对痰液脱落细胞进行荧光原位杂交(fluorescence in situ hybridization,FISH)用以分析肺癌的染色体畸变,探讨FISH技术辅助诊断肺癌的可行性和有效性。方法选用3、7、17号染色体着丝粒探针,p16、p53探针对40例疑似肺癌患者痰液的脱落细胞进行FISH研究。结果肺癌患者痰液脱落细胞中3、7、17号染色体数目和p16、p53畸变阳性率分别为41.9%、45.2%、41.9%、54.9%、51.6%,其中p16、p53畸变率较高。FISH联合应用五种探针诊断肺癌的敏感性为80.6%,特异性为77.8%,诊断效率为80.0%。结论肺癌的发生、发展与染色体的畸变有关,应用FISH技术检测肺癌患者痰液细胞染色体数目畸变,可作为肺癌诊断的一项辅助方法。     

Expression of the epithelial L1 antigen as an immunohistochemical marker of squamous cell carcinoma of the lung

The distribution of epithelial L1 antigen was evaluated in 139 bronchogenic carcinomas which had been classified by a panel of pathologists according to the WHO recommendation of 1981. L1 was not found in three large cell and 13 small cell carcinomas, but it was expressed by tumour cells in 67 of 69 squamous cell carcinomas (97%), in three of four adenosquamous carcinomas (75%), and in three of 49 adenocarcinomas (6%). The staining for L1 antigen was more diffusely distributed in the positive adenocarcinomas than in the squamous cell carcinomas. Its expression in squamous cell carcinomas was typically confined to relatively small tumour cell groups and never included a complete specimen. Semi-quantitative estimation of the immunostaining showed no clear relationship to the degree of differentiation and scores for proliferation, but L1 expression was negatively related to nuclear aberration (P less than 0.025) and malignancy scores (P less than 0.002). The good agreement between morphological classification and expression of L1 makes this a valuable marker in the diagnosis of lung carcinomas.     

AQUA and FISH analysis of HER-2/neu expression and amplification in a small cell lung carcinoma tissue microarray

AIMS: Most small cell lung carcinoma (SCLC) patients have metastatic disease at the time of diagnosis and are faced with poor prognosis and limited treatment options. Reports of HER-2/neu gene amplification and overexpression in this malignancy have raised the possibility of applying targeted immunotherapy with trastuzumab, the monoclonal antibody used to treat metastatic breast cancer. However, a review of the studies measuring HER-2/neu gene amplification and protein expression in SCLC reveals discordant results. The aim of the present study was to re-examine HER-2/neu expression in SCLC in relation to gene copy number using the new, highly sensitive, immunofluorescence automated quantitative analysis (AQUA) technology. METHODS AND RESULTS: Fluorescence in situ hybridization (FISH) was used to measure HER-2/neu gene copy number and amplification status and AQUA was used to measure protein expression in a series of 23 SCLC tumours on a tissue microarray. None of the 17 SCLC specimens assessable by FISH exhibited HER-2/neu gene amplification as defined by a HER-2/neu/chromosome 17 ratio = or > 2. Twelve of 17 (70.1%) SCLC samples were polysomic for chromosome 17 with corresponding increases in HER-2/neu gene copy numbers. Intermediate levels of protein expression corresponding to AQUA scores in the range of 4-24 were detected in all 23 specimens. High protein expression levels corresponding to AQUA scores up to 83, observed previously in association with gene amplification and poor prognosis in breast cancer cases, were not detected in the present study. No statistically significant association was observed between absolute chromosome 17 or HER-2/neu gene copy numbers and protein expression levels in tumour cells (P > 0.45). CONCLUSIONS: The lack of gene amplification and robust HER-2/neu protein expression in SCLC tumour cells in this series does not suggest a prominent role for the HER-2/neu gene in SCLC tumour progression and does not support the general applicability of targeted immunotherapy with trastuzumab to this malignancy.     

免疫组织化学和荧光原位杂交检测肺腺癌ROS1表达的对比

目的比较免疫组织化学(immunohistochemistry,IHC)和荧光原位杂交(fluorescent in situhybridization,FISH)技术在检测肺腺癌中ROS1蛋白表达和融合基因的差异性和一致性,评估IHC在临床上的应用价值。方法应用IHC和FISH技术对332例肺腺癌中ROS1蛋白表达和融合基因进行检测,比较两者检测的结果和相关性,并探讨ROS1融合基因与临床病理特征之间的关系。结果 332例肺腺癌中FISH阳性者13例,阴性者319例,ROS1融合基因阳性率为3.9%;ROS1蛋白表达结果:0者312例,1+者2例,2+者5例,3+者13例,ROS1蛋白过表达率为6.0%;经FISH检测312例ROS1蛋白表达为0的标本无ROS1融合基因,2例1+的标本中也无ROS1融合基因;5例2+的标本中有2例ROS1融合基因,13例3+的标本中11例有ROS1融合基因;IHC检测0和1+、2+、3+标本FISH检测阳性符合率分别为0(0/314)、40%(2/5)和84.6%(11/13);IHC检测ROS1蛋白为2+~3+的标本中72.2%(13/18)显示ROS1融合基因,0~1+患者中无1例有ROS1融合基因(Kappa系数=0.831,P0.01);IHC检测ROS1蛋白表达的敏感性和特异性分别100%和98.4%。结论 IHC检测ROS1蛋白表达和FISH检测ROS1融合基因,两种方法有较高的符合率,一致性较好,IHC有较高的敏感性和特异性,可作为临床检测肺腺癌ROS1融合基因的简单而有效的方法。