EB病毒基因在喉鳞状细胞癌中的表达

目的：ＥＢ病毒感染与喉鳞状细胞的发生发展是否有关迄今未明。本文目的在于了解ＥＢ病毒基因在喉鳞状细胞癌组织中的表达情况，探讨各种ＥＢ病毒基因产物在喉鳞状细胞癌发生发展中的生物学意义。方法：采用原位杂交和免疫组化技术，检测２９例不同分化类型喉鳞状细胞癌组织中ＥＢ病毒编码的小ＲＮＡ（ＥＢＥＲｓ）、潜伏感染膜蛋白（ＬＭＰ１）、ＥＢ病毒核抗原（ＥＢＮＡ－１）、溶解感染立即早期基因编码蛋白ＺＥＢＲＡ、早期基因编码蛋白ＥＡ－Ｄ、晚期基因编码蛋白ＶＣＡ和ＭＡ。结果：各检测指标在未分化癌、低分化和中分化鳞癌均有一定比例的阳性，但在２例高分化鳞癌均为阴性。发现ＥＢ病毒潜伏性和溶解性感染基因产物表达分别随着癌分化程度增高而减弱和增强。７例低分化和１例中分化鳞癌ＬＭＰ－１除胞膜和胞浆阳性外，胞核也呈明显阳性。２例低分化和１例中分化鳞癌中有少量癌细胞ＺＥＢＲＡ阳性。结论：部分喉鳞状细胞癌尤其是未分化和低分化鳞癌的发生发展与ＥＢ病毒感染密切相关，提示癌细胞的分化程度与ＥＢ病毒基因表达的差异有关。     

大肠癌中C－erbB－2癌基因产物的表达及意义

应用ＡＢＣ免疫组化方法和Ｃ－ｅｒｂＢ－２癌基因的单克隆抗体对９３例大肠组织标本进行了免疫组化染色。结果表明：在６８例大肠癌中，Ｃ－ｅｒｂＢ－２阳性表达为６１．８％。其中高分化腺癌组Ｃ－ｅｒｂＢ－２阳性表达为４０％；中分化腺癌组阳性表达为５０％。低分化腺癌组阳性表达为８１．２％；粘液细胞癌组阳性表达为８５．７％。粘液细胞癌组和低分化癌组Ｃ－ｅｒＢ－２阳性表达明显高于其它２组（Ｐ＜０．０１）。在已有淋巴结转移的大肠癌中，Ｃ－ｅｒｂＢ－２癌基因阳性表达为７６．２％（３２／４２）。在无淋巴结转移的大肠癌中，Ｃ－ｅｒｂＢ－２阳性表达为３８．５％（１０／２６）。二者比较，Ｃ－ｅｒｂＢ－２癌基因表达有显著性差异（Ｐ＜０．０１）。在１５例大肠腺瘤标本中，Ｃ－ｅｒｂＢ－２癌基因阳性表达为６．７％。１０例慢性炎症和１０例正常肠粘膜Ｃ－ｅｒｂＢ－２则均为阴性。以上这些结果提示：Ｃ－ｅｒｂＢ－２癌基因产物不仅可视为肿瘤的一种标志，而且与大肠癌的分化程度、淋巴结的转移密切相关，这证明Ｃ－ｅｒｂＢ－２癌基因的检测对大肠癌的病理诊断和预后判断有很高的实用价值。     

C－erbB＿2、p53、p21在卵巢粘液性癌表达及其临床意义的研究

本实验对５０例卵巢粘液性囊腺癌中Ｃ－ｅｒｂＢ２（ｎｅｕ）癌基因，ｐ５３抗癌基因、ｐ２１蛋白（ｒａｓ基因产物）的表达进行研究，观察其在卵巢癌、交界性瘤、良性瘤的表达率，并采用半定量的ＡＢＣ免疫组化染色方法，研究不同强度表达与病理分化、临床分期、预后关系，得到初步结果。Ｃ－ｅｒｂＢ２在卵巢粘液癌中表达４８％，其限性表达率及表达强度与预后和病理分化有关。ｐ５３在卵巢粘液癌中表达４Ｏ％，但其表达率、表达强度与预后、手术分期和病理分化无明显相关性。ｐ２１在粘液癌中表达１２％，其表达率及强度与预后和病理分化无明显关系。在交界瘤、良性瘤中无表达。     

胃癌组织中肥大细胞与C－erbB－2癌基因蛋白

用俾士麦棕法和免疫组织化学方法检测了５７例胃癌组织中肥大细胞（ＭＣ）和Ｃ－ｅｒｂＢ－２癌基因蛋白。结果表明：（１）ＭＣ计数与胃癌的分化程度和转移有关，高、中分化腺癌高于低分化腺癌和未分化癌（Ｐ＜０．０５），无转移者高于有转移者（Ｐ＜０．０５）；（２）Ｃ－ｅｒｂＢ－２癌基因蛋白阳性表达与胃癌的分化程度和转移无关（Ｐ＞０．０５）；（３）Ｃ－ｅｒｂＢ－２癌基因蛋白阳性表达与ＭＣ计数有关，ＭＣ高计组，Ｃ－ｅｒｂＢ２癌基因蛋白阳性表达低于ＭＣ低计组（Ｐ＜０．０５）。     

乳腺肿瘤中C—erbB—2癌基因蛋白的表达及意义

目的：探讨Ｃ－ｅｒｂＢ－２癌基因蛋白在乳腺中肿瘤中的表达情况及临床意义。方法：应用免疫组化ＳＰ法,对１２６例乳腺良恶性肿瘤进行Ｃ－ｅｒｂＢ－２癌基因蛋白检测,其中９７例乳腺癌进行激素受体（ＥＲ）和孕激素受体（ＰＲ）检测。结果：Ｃ－ｅｒｂＢ－２蛋白阳性表达的细胞主要为癌细胞。Ｃ－ｅｒｂＢ－２蛋白在导管内癌、Ｐａｇｅｔ‘ｓ蛋白在导管内癌Ｐａｇｅｔ’ｓ病是浸润性导管癌中呈高表达,在浸润的小叶癌中均呈阴性     

谷胱甘肽S转移酶在转移性肺癌的表达

酸性谷胱甘肽Ｓ转移酶（ＧＳＴ－π）在大肠癌、胃癌、肺癌、食管癌及乳腺癌中表达较高，并可作为非小细胞肺癌（ＮＳＣＬＣ）耐药性标志［１，２］。近来研究表明一些耐药基因在复发转移癌中表达较原发病灶明显增高。本文应用ＲＴ－ＰＣＲ技术检测５２例非小细胞肺癌ＧＳ...     

Fos、Jun、Rb、p16和bc1-2蛋白在食管癌的表达及其意义

目的　研究同一食管癌组织中癌基因产物和抗癌基因产物的表达,并探讨其与食管癌的发生和发展及生物学行为的关系。方法　应用免疫组织化学ＳＡＢＣ 法,观察４２ 例食管癌组织和５ 例切缘食管粘膜中Ｆｏｓ、Ｊｕｎ 、Ｒｂ、ｐ１６ 和ｂｃ１ －２ 蛋白的表达,并比较其阳性率。结果　４２ 例食管癌组织中Ｆｏｓ、Ｊｕｎ、Ｒｂ 、ｐ１６、ｂｃ１－ ２ 阳性检出率分别为５９－５％ 、５０－０％ 、８５－７ ％ 、６４－３ ％ 和３８－１％ ;Ｆｏｓ、Ｊｕｎ 、Ｒｂ 、ｐ１６ 和ｂｃ１ － ２ 的阳性表达率分别与食管癌的分化程度和淋巴结转移关系密切（ Ｐ＜ ０ ．０５） 。结论　提示多种癌基因蛋白均参与食管癌的发生和发展过程,有不同的生物学作用。联合检测癌基因蛋白有助于了解食管癌病人的预后。     

食管鳞状细胞癌中Rb和p16蛋白表达的免疫组织化学研究

目的探讨与细胞周期调节有关的基因改变在食管鳞状细胞癌中的表达及其意义。方法应用免疫组织化学链霉抗生物素蛋白过氧化物酶连接法（ＳＰ法），检测７３例食管癌中Ｒｂ蛋白和ｐ１６蛋白的表达。结果食管鳞状细胞癌Ｒｂ和ｐ１６蛋白表达阳性率分别为４２５％（３１／７３）和６７１％（４９／７３），Ｒｂ蛋白表达阳性率与食管鳞状细胞癌分级呈显著负相关（Ｐ＜０００５），ｐ１６蛋白表达阳性率与食管鳞状细胞癌分级呈显著正相关（Ｐ＜０００５）。４９例ｐ１６蛋白表达阳性的食管癌中Ｒｂ蛋白表达阴性者３９例，３１例Ｒｂ蛋白表达阳性的食管癌中ｐ１６蛋白表达阴性者２１例，两者呈显著负相关（Ｐ＜０００５）；１０例食管鳞状细胞癌呈Ｒｂ蛋白和ｐ１６蛋白共同表达，３例Ｒｂ蛋白和ｐ１６蛋白表达呈共同阴性。结论Ｒｂ蛋白和ｐ１６蛋白表达的有无可分别作为食管鳞状细胞癌病理分级参考指标之一，并间接证明Ｒｂ蛋白对ｐ１６蛋白表达具有负性调节的功能。     

大肠癌中C—erbB—2癌基因产物的表达及意义

应用ＡＢＣ免疫组化方法和Ｃ－ｅｒｂＢ－２癌基因的单克隆抗体为９３例大肠组织标本进行了免疫组化染色。结果提示：Ｃ－ｅｒｂＢ－２癌基因产物不仅可视为肿瘤的一种标志，而且与大肠癌的分化程度、淋巴结的转移密切相关，这说明Ｃ－ｅｒｂＢ－２癌基因的检测对大肠癌的病理诊断和预后判断有很高的实用价值。     

胃癌多基因表达的同步检测

应用ＬＳＡＢ和ＡＢＣ法对７５例胃癌及癌旁组织进行了ｐ５３、ｃ－ｅｒｂＢ－２、ＥＧＦＲ和ｒａｓ表达的研究。结果显示：（１）７５例胃癌ｐ５３、ｃ－ｅｒｂＢ－２、ＥＧＦＲ和ｒａｓ表达阳性率分别为４１．３％、１８．７％、６１．３％和４６．７％。ｐ５３在肠型胃癌中阳性率为５２．６％，高于弥漫型胃癌的２９．７％（Ｐ＜０．０５）；在早期胃癌中阳性率较高（６０．０％），癌旁重度异型增生亦有阳性表达（２／４）。ｃ－ｅｒｂＢ－２阳性表达只限于癌灶，而癌旁组织均为阴性。ｃ－ｅｒｂＢ－２在高分化胃癌中阳性率较高（Ｐ＜０．０５）。ＥＧＦＲ表达与胃癌的大体类型、生长方式、分化程度、淋巴结受累和远处转移呈正相关（Ｐ＜０．０５）。（２）ｃ－ｅｒｂＢ－２和ＥＧＦＲ在胃癌中的表达互相独立。（３）ｒａｓ表达与ＥＧＦＲ表达呈正相关，而与ｃ－ｅｒｂＢ－２呈负相关。（４）ｐ５３表达与其它基因表达无明显关系。上述结果表明，胃癌发生发展过程中伴有多种癌基因的变化，其出现时间不同，意义也不一样。     

Differential expression of bcl2 protooncogene in neuroblastoma and other human tumor cell lines of neural origin.

The bcl2 protooncogene was originally discovered because of its involvement in t(14;18) chromosomal translocations frequently found in non-Hodgkin's lymphomas. The expression of this gene is reported to be highly tissue specific, with bcl2 mRNAs being readily detectable only in hematolymphoid tissues and brain. To explore the possible involvement of bcl2 in neural tumors, we surveyed a variety of tumor cell lines for the presence of the p26-BCL2 protein by immunoprecipitation and immunoblotting methods. Very high levels of BCL2 protein were found in three of nine neuroblastoma (NB) cell lines examined; these levels of p26-BCL2 were comparable to lymphoma cell lines that contain a t(14;18). Despite the impressive relative amounts of BCL2 protein, however, no structural alterations or changes in the methylation status of bcl2 genes were detected in these NB cell lines by conventional Southern blotting. Of the other NB cell lines surveyed, three contained intermediate levels of BCL2 and another three cell lines had little or no detectable BCL2 protein, raising the possibility that determination of relative levels of BCL2 protein may help to segregate neuroblastomas into groups with different biological and clinical characteristics. BCL2 protein levels were not influenced by induction of neuronal differentiation with nerve growth factor in two of the two cell lines examined [SH-SY5Y (high BCL2); GICAN (low BCL2)] and did not correlate with N-MYC gene amplification or expression of nerve growth factor receptors. NB cell lines that contained little or no detectable BCL2 protein, however, tended to contain significant proportions of flat epithelioid cells, whereas bcl2-expressing cell lines were composed primarily of neuronal-like cells, suggesting that expression of this protooncogene correlates with the differentiation characteristics of these tumor cell lines. In addition to NBs, lower levels of BCL2 protein were also found in a variety of other neural crest-derived tumors and tumor cell lines, including some neuroepitheliomas, Ewing's sarcomas, neurofibromas, and melanomas. With regard to tumors of central nervous system origin, bcl2 expression was absent from most medulloblastomas but was detected at moderate to low levels in a retinoblastoma and some glioblastoma multiforme cell lines. Taken together, these findings imply that bcl2 protooncogene expression is differentially regulated within the various lineages of cells that give rise to the nervous system.     

BCL2 overexpression in diffuse large B-cell lymphoma.

Translocation (14;18)(q32;q21), which is detected in 20-30% of diffuse large B-cell lymphomas (DLBCL), is regarded as a major mechanism for BCL2 protein overexpression. Nevertheless, BCL2 overexpression is not always caused by t(14;18), because it is often detected in lymphomas without BCL2 rearrangement. Recent studies have shown that increased expression of BCL2 may also result from BCL2 gene amplification in DLBCL. Similarly, it has been speculated that the mutations of the open reading frame might cause increased expression of BCL2 by affecting the interactions of BCL2 with other proteins. The results obtained from studies on the association of BCL2 protein overexpression with survival of DLBCL are controversial, although a correlation with decreased overall survival seems to exist. However, BCL2 rearrangement does not seem to have any major association with poor prognosis, but it is difficult to assess its true impact on prognosis due to differences in treatment and follow-up, and methodologies applied to study the BCL2 rearrangement. This review summarizes the BCL2 expression studies in DLBCL and discusses the prognostic relevance of BCL2 overexpression and its mechanisms.     

The prognostic impact of BCL2 protein expression in acute myelogenous leukemia varies with cytogenetics.

BCL2 protein exerts an antiapoptotic effect on cells and decreases chemosensitivity. To determine whether BCL2 expression is prognostic of patient outcome in acute myelogenous leukemia (AML), we measured its level in 198 newly diagnosed, untreated AML patients by Western blotting using whole-cell lysates from low-density peripheral blood cells. BCL2 expression was not associated with the percentage of blasts (R2 = 0.10), French-American-British classification type, or cytogenetic abnormality. Smoothed martingale residual plots indicated that high BCL2 protein level was an adverse prognostic factor for patients with either favorable or intermediate prognosis cytogenetics [FIPC; inv(16), t(8:21), t(15:17), or diploid or insufficient metaphases] but was a favorable prognostic factor for patients with unfavorable prognosis cytogenetics (UC; -5, -7, +8, 11q23, Ph1, or miscellaneous changes). Patients with FIPC and high BCL2 (highest quartile) had a significantly shorter median survival (78 weeks versus not reached; P = 0.009) than did those with lower (lower three quartiles) levels of BCL2. Among those with UC, as BCL2 level decreased from the fourth quartile to the third or the combined first and second quartiles, the median survival decreased (from 94 to 45 or 17 weeks, respectively; P = 0.003). Lower expression of BCL2 in UC was associated with shorter remission duration (P = 0.05). In multivariate analyses performed using either overall or event-free survival as the end point, for either all patients or within either cytogenetic subgroup, BCL2 level was an independent prognostic factor. Similar analysis revealed that BCL2 level was an independent predictor of remission duration for UC patients as well. These data suggest that BCL2 is involved differently in different types (favorable versus unfavorable) of AML and that therapeutic strategies aimed at modulating BCL2 function may be more likely to work in patients with favorable cytogenetic abnormalities.     

The t(14;18) is associated with germinal center-derived diffuse large B-cell lymphoma and is a strong predictor of outcome.

The t(14;18) is present in a significant proportion of diffuse large B-cell lymphomas (DLBCLs), however, the prognostic effect of the translocation and the relationship with transformed follicular lymphoma remains controversial. To clarify these uncertainties, interphase fluorescence in situ hybridization (FISH) was used to determine the incidence of the t(14;18) in nodal DLBCL, and this was correlated with BCL2 expression, germinal center (GC) immunophenotype, and patient outcome. FISH was performed on paraffin-extracted nuclei from 137 de novo nodal DLBCLs. Eighteen of 137 de novo DLBCLs were t(14;18) positive. The t(14;18) was most commonly associated with the subset of DLBCLs that expressed a GC phenotype, defined as CD10+, BCL6+ (GC-type DLBCL): positive in 14 of 47 (30%) cases, compared with 4 of 89 (5%) in the non-GC group (Pearson's chi(2) = 28.4; P < 0.0001). All cases with a translocation expressed BCL2 protein, however, 40 expressed BCL2 protein without a t(14;18). GC-type DLBCL patients with a t(14;18) had a significantly worse survival compared with GC-type DLBCL patients without the translocation (2-year survivals were 29 and 63%, respectively; P = 0.006). Of the cases without the translocation, BCL2 protein expression did not affect survival. In contrast, in the non-GC group of DLBCLs, BCL2 protein expression reduced the 2-year overall survival from 64% in the BCL2-negative group to 38%, with a median survival of 15.0 months (P = 0.02). In conclusion, the t(14;18) is common in DLBCLs, particularly in GC-type DLBCLs, where the presence of the translocation has a poor prognostic effect. BCL2 protein expression defines a group of non-GC DLBCL patients with a poor prognosis.     

Molecular and immunological dissection of diffuse large B cell lymphoma: CD5+, and CD5- with CD10+ groups may constitute clinically relevant subtypes.

Diffuse large B cell lymphoma (DLBL) constitutes the greatest percentage of adult non-Hodgkin's lymphomas and represents a diverse spectrum of lymphoid neoplasms. Clinicopathologic, phenotypic and genotypic findings were correlated and compared for 63 DLBL cases to investigate whether they represent clinically relevant subtypes. They were all cyclin D1 negative and were phenotypically divided into three groups, ie group I (CD5+ type, n=11), group II (CD5- CD10+ type, n=19), and group III (CD5- CD10- type, n=33). Data were correlated by observing the respective gene rearrangement and expression of BCL2 and BCL6. In clinical aspects, the group I cases demonstrated a significantly inferior survival than those of the other two groups (log-rank test, P = 0.016). Although rearrangement of BCL2 and BCL6 did not show any inclination to a specific subgroup, the immunohistochemical detection of BCL2 was less frequent, at a statistically significant level (P=0.011), in group II (50%) than in group I (82%) and III (82%) cases. This appears to confirm the unique aspect of the CD5- CD10+ type DLBL, indicating a certain relationship with the normal germinal center cells which usually lack BCL2 expression. The BCL6 protein expression was detected in most of the present DLBL cases (92%) irrespective of this grouping. These data suggest that the phenotypic delineation by the detection of CD5 and CD10 will improve our understanding of DLBL and be helpful in a future subgrouping of DLBL.     

Altered expression of the retinoblastoma gene product: prognostic indicator in bladder cancer.

BACKGROUND: It has been reported that 50%-70% of patients with bladder cancer experience recurrence after initial successful treatment and about 10%-20% of these patients die of the disease. Despite precise pathologic staging and grading, we are unable to predict clinical outcome in all patients. The retinoblastoma-susceptibility (RB) gene, a prototype of tumor suppressor genes, has recently been associated with development and/or progression of bladder cancer, as well as sarcoma and small-cell lung cancer. In transitional cell carcinomas of the bladder, we have observed altered expression of the Rb gene product--a nuclear phosphoprotein thought to function as a cell cycle regulator. PURPOSE: The aim of this study was to investigate the hypothesis that altered patterns of Rb expression correlate with prognosis in bladder cancer. METHODS: Expression of the RB gene was evaluated in specimens from 48 primary bladder tumors obtained by cystectomy or transurethral resection. Rb protein expression was correlated with disease outcome in these patients. Rb expression was examined by immunohistochemistry, using the mouse monoclonal antibody Rb-PMG3-245 on frozen tissue sections. Computerized image analysis was used to quantify the level of Rb protein in individual tumor cells. RESULTS: The overall 5-year disease-free survival was 66%, with a median follow-up of 42 months. Normal levels of Rb protein expression were found in 34 patients (Rb-positive group). A spectrum of altered patterns of expression from undetectable levels to heterogeneous expression, however, was observed in 14 patients (altered Rb group). Of the 38 patients with muscle-invasive tumors, 13 were categorized as having altered expression of Rb protein. Only one of 10 patients with superficial carcinomas had altered expression of Rb protein. The 5-year survival was significantly decreased in patients with altered Rb protein compared with the survival in patients with positive Rb expression (P less than .001). CONCLUSIONS: The results suggest that tumors exhibiting decreased expression of the RB gene-coded product (Rb protein) had a more aggressive biological behavior than those that expressed the Rb protein in the majority of their tumor cells. IMPLICATIONS: This study demonstrates that altered patterns of Rb protein expression may be an important prognostic variable in patients presenting with invasive bladder cancer.     

Immunohistochemical double-hit score is a strong predictor of outcome in patients with diffuse large B-cell lymphoma treated with rituximab plus cyclophosphamide, Doxorubicin, vincristine, and prednisone

PURPOSE Approximately 5% of diffuse large B-cell lymphomas (DLBCLs) are double-hit lymphomas (DHLs) with translocations of both MYC and BCL2. DHLs are characterized by poor outcome. We tested whether DLBCLs with high expression of MYC protein and BCL2 protein share the clinical features and poor prognosis of DHLs. PATIENTS AND METHODS Paraffin-embedded lymphoma samples from 193 patients with de novo DLBCL who were uniformly treated with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) were studied using immunohistochemistry for MYC, BCL2, CD10, BCL6, and MUM1/interferon regulatory factor 4, and fluorescent in situ hybridization (FISH) for MYC and BCL2. Results FISH analysis identified DHL in 6% of patients, who showed the expected poor overall survival (OS; P = .002). On the basis of immunohistochemical MYC and BCL2 expression, a double-hit score (DHS) was assigned to all patients with DLBCL. The DHS-2 group, defined by high expression of both MYC and BCL2 protein, comprised 29% of the patients. DHS 2 was significantly associated with lower complete response rate (P = .004), shorter OS (P < .001), and shorter progression-free survival (PFS; P < .001). The highly significant correlation with OS and PFS was maintained in multivariate models that controlled for the International Prognostic Index and the cell-of-origin subtype (OS, P < .001; PFS, P < .001). DHS was validated in an independent cohort of 116 patients who were treated with R-CHOP. CONCLUSION The immunohistochemical DHS defined a large subset of DLBCLs with double-hit biology and was strongly associated with poor outcome in patients treated with R-CHOP.     

BCL2 expression correlates with metastatic potential in pancreatic cancer cell lines

BACKGROUND: Programmed cell death (termed apoptosis) regulates normal tissue homeostasis. Loss of local paracrine signals and intercellular adhesion molecules are potent inducers of apoptosis and thereby eliminate normal cells that may have escaped beyond the confines of the local organ environment. Dysregulation in the expression of the BCL2 gene family, the prototypic regulators of apoptosis, is a common occurrence in cancer and imparts resistance to standard triggers of apoptosis. Therefore, the authors sought to examine whether abnormal BCL2 gene family expression correlated with resistance to apoptosis and increased metastatic potential in pancreatic carcinoma. METHODS: The authors examined BCL2 expression and apoptotic sensitivity in three panels of human pancreatic cancer cell lines that possess varying metastatic potential. Stable transfectants were generated that overexpress BCL2. These transfectants were then analyzed for differences in metastasis formation in athymic mice. RESULTS: Among the isogenic panels of pancreatic cancer cell lines, BCL2 expression levels correlated with metastatic potential. Highly metastatic variants of each family of cell lines were more resistant to induction of apoptosis. Finally, using the BCL2 transfectant in a xenograft model, elevated BCL2 expression led to a higher incidence of metastases. CONCLUSIONS: The authors conclude that increased BCL2 expression correlates with apoptotic resistance and metastatic potential; dysregulation of BCL2 expression may be involved in the metastatic progression of pancreatic carcinoma.