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1.
The accurate assessment ofa proto-oncogene, human epidermal growth factor receptor-2 gene (HER-2), is extremely important for the therapy and prognosis of breast cancer. Currently, immunohistochemistry (IHC) is the method widely used for the detection of HER-2 protein. Fluorescence in situ hybridization (FISH) has been suggested to be a golden standard assay for HER-2 amplification. This study examined the expression and amplification of HER-2 in paraffin-embedded sections of breast cancer tissues, and compared the two methods on the measurement of HER-2 status. HER-2 gene and protein were determined in breast cancer samples from 52 Chinese women by FISH and IHC respectively. The findings indicated that the HER-2 gene amplification was found in 18 cases (34.6%) by FISH and the HER-2 protein over-expression (score 3+) in 15 cases (28.8%) by IHC. hnmunohistochemically, 28.6% of the cases scored as 2+ and 93.3% of the cases scored as 3+ were HER-2-positive by FISH. There was a significant correlation between the HER-2 gene amplification and HER-2 protein over-expression in breast cancer (P〈0.005). No correlation was noted between the HER-2 gene amplification and any of the clinicopathological parameters examined, including age, menopausal status, menarche age, tumor size, histological tumor type, histological grade, lymph node status, and the expression of ER and PR. It was concluded that the detection of HER-2 gene amplification in breast cancer by FISH is valuable and can compare with HER-2 protein detection by IHC.  相似文献   

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OBJECTIVE The P53 tumor suppressor gene shows the most frequent genetic alteration in human tumors. Mutation, deletion and rearrangement of P53 gene have been found in several types of neoplasm including osteosarcoma. The present study is to clarify the status of abnormalities of this gene in osteosarcoma in China.
METHODS Forty-six osteosarcomas were collected from 32 male and 14 female patients aged 20.6 years on average. Eight patients had lung metastasis. Immunohistochemistry, Southern blot and contrast flow cytometry were used for the study.
RESULTS Twenty-seven of 46 (58.7%) osteosarcoma specimens showed strongly positive reaction to the MoAb BP53-12 staining (+2 - +3), which indicated overexpression of P53 protein in osteosarcoma. Contrast study with DNA flow cytometry made on osteosarcoma showed that most of the tumors with strongly positive P53 have higher DNA content than those of negative ones. Alterations of the restriction pattern of the P53 gene were detected in 4 of 20 osteosarcomas in the Hind III group and 3 of 24 in the Eco R I group. Three of them were part or whole deletion of the gene and five of them had extraband which indicated rearrangement of the gene.
CONCLUSIONS The results of this study showed that overexpression of P53 oncoprotein is one of the most frequent genetic changes in osteosarcoma and P53 oncoprotein expression analysis could be a prognostic parameter in osteosarcoma. P53 gene abnormalities play an important role in the development of transformation and proliferation of osteosarcoma cells.
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BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most prevalent fatal cancers in the world. Despite advances in early diagnosis and improvements in surgical techniques, the survival of patients with HCC even after resection is poor because of the high incidence of recurrences. Therefore, the identification of prognostic factors may be helpful in the development of new treatment protocols. AIMS: To investigate HER-2/neu status in HCC by immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH), and to explore the possibility of using trastuzumab in the treatment of HCC. METH ODS: Eight hundred and sixty eight surgical samples from patients with primary HCC were examined for their HER-2/neu status. IHC for HER-2/neu was performed with the HercepTest kit; FISH analysis was performed with the PathVysion HER-2 DNA probe kit. The correlations between HER-2/neu overexpression and clinicopathological characteristics were analysed statistically. RESULTS: HER-2/neu overexpression was detected in 21 (2.42%) of the 868 primary HCCs. Only one specimen showed HER-2/neu gene amplification by FISH. No significant associations were found between HER-2/neu overexpression and the clinicopathological parameters. CONCLUSIONS: There is a low frequency of HER-2/neu overexpression/amplification in HCC. There appears to be no role for HER-2/neu as a prognostic marker and no benefit of anti-HER-2/neu trastuzumab treatment in patients with HCC.  相似文献   

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Background: Hepatocellular carcinoma (HCC) is one of the most prevalent fatal cancers in the world. Despite advances in early diagnosis and improvements in surgical techniques, the survival of patients with HCC even after resection is poor because of the high incidence of recurrences. Therefore, the identification of prognostic factors may be helpful in the development of new treatment protocols. Aims: To investigate HER-2/neu status in HCC by immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH), and to explore the possibility of using trastuzumab in the treatment of HCC. Methods.. Eight hundred and sixty eight surgical samples from patients with primary HCC were examined for their HER-2/neu status. IHC for HER-2/neu was performed with the HercepTest kit; FISH analysis was performed with the PathVysion HER-2 DNA probe kit. The correlations between HER-2/neu overexpression and clinicopathological characteristics were analysed statistically. Results: HER-2/neu overexpression was detected in 21 (2.42%) of the 868 primary HCCs. Only one specimen showed HER-2/neu gene amplification by FISH. No significant associations were found between HER-2/neu overexpression and the clinicopathological parameters. Conclusions: There is a low frequency of HER-2/neu overexpression/amplification in HCC. There appears to be no role for HER-2/neu as a prognostic marker and no benefit of anti-HER-2/neu trastuzumab treatment in patients with HCC.  相似文献   

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Studies have proved that microRNA-101 (miR-101) functions as a tumor suppressor and is associated with growth and apoptosis of various human cancers. However, the role of miR-101 in os- teosarcoma and the possible mechanism by which miR-101 affects the tumor growth and apoptusis have not been fully elucidated. In this study, we found that the expression ofmiR-101 was down-regulated in osteosarcoma tissues and Saos-2 cell line as compared with that in adjacent non-neoplastic bone tissues and the osteoblastic cell line. To better characterize the role of miR-101 in osteosarcoma, we used a gain-of-function analysis by transfecting human osteosarcoma cell line Saos-2 with chemically synthe- sized miR-101 mimics. The results showed that overexpression of miR-101 inhibited the proliferation and promoted the apoptosis of Saos-2 cells. Meanwhile, bioinformatic analysis demonstrated that mTOR gene was a direct target of miR- 101. Overexpression of miR- 101 significantly decreased the ex- pression of roTOR at both mRNA and protein levels in Saos-2 cells, consequently inhibiting Saos-2 cells proliferation and promoting ceils apoptosis in an mTOR-dependent manner. Taken together, these data suggest that miR-101 may act as a tumor suppressor, which is commonly downregulated in both osteosarcoma tissues and cells, roTOR plays an important role in mediating miR-101 dependent bio- logical functions in osteosarcoma. Reintroduction of miR-101 may be a novel therapeutic strategy bydown-regulating mTOR expression.  相似文献   

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Background Accurate detection of human epidermal growth factor receptor 2 (HER2) expression and gene amplification is crucial for the application of HER2-specific therapy and for evaluating the response of patients with breast cancer.A uniform and standard procedure of immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) needs to be established for evaluating the HER2 status in breast cancer tissues for the treatment of patients with real HER2-positive tumors.The present multicenter study was aimed to examine the HER2 status in breast cancer specimens from Chinese patients using both IHC and FISH methods.Methods A multicenter study was performed on the HER2 status in 3 149 breast cancer specimens from different ethnic populations and areas in China by IHC and FISH assays.The potential association of HER2 status with demographic and clinical characteristics was analyzed.Results The positive rates for HER2 over-expression and HER2 amplification were 23.3% and 27.5% in this study,respectively.The concordance between IHC and FISH was 71.2% (K=0.494,P <0.001).Furthermore,72.9% of specimens with IHC 2+ were negative to FISH.The discordance rates among laboratories were from 5% to 28% for IHC and 1% to 16% for FISH.HER2 amplification was associated significantly with advanced tumor stage (Ⅲ or Ⅳ,P=0.002),large tumor size (>5 cm,P=0.002),moderate and poor histological grades (P <0.0001),post-menopause (P <0.0001),ER-PR-(P=0.002),and having >4 lymph nodes affected (P <0.0001) in this population.The positive rates of HER2 amplification in specimens from Man and Hui Chinese were significantly higher than that in other Chinese populations.There are slightly higher positive rates of HER2 expression and amplification in Chinese patients with breast cancer.Conclusion These findings may provide new insights into understanding the epidemiological features of HER2 expression and amplification,and may be valuable for clinical practice.  相似文献   

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乳腺癌HER-2的表达与临床病理特征的关系   总被引:1,自引:0,他引:1  
Li HH  Ma F  Zeng X  Wang JY  Yuan P  Fan Y  Xu BH 《中华医学杂志》2011,91(2):76-80
目的 探讨荧光原位杂交(FISH)和免疫组织化学(IHC)检测乳腺癌组织中人表皮生长因子受体2(HER-2)基因扩增及蛋白表达的一致性和相关性,及其与乳腺癌患者的临床病理特征的关系.方法 采用FISH法检测128例乳腺癌患者HER-2基因扩增状态,与IHC结果进行一致性及相关性分析,并比较其与乳腺癌患者的临床病理特征的相关性.结果 128例乳腺癌标本中,IHC与FISH结果符合率为90.6%,存在一致性(Kappa=0.405,P=0.000).两种检查结果呈正相关(r=0.655,P=0.000).ER表达与HER-2基因扩增及其蛋白表达状态呈负相关(r=-0.300,P=0.001;r=-0.223,P=0.011),而ER/PR状态与HER-2基因扩增状态呈负相关(r=-0.213,P=0.016).肿瘤分级与HER-2蛋白表达呈负相关(r=-0.293,P=0.008),而与HER-2基因扩增状态无关(P>0.05).结论 IHC(+++)与基因扩增有较好的一致性,而IHC(+~++)者有必要进一步行FISH法检测基因状态.ER、ER/PR状态及肿瘤分级与HER-2基因扩增和(或)蛋白表达存在相关性.
Abstract:
Objective To investigate the concordance and correlation between fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) assessment for HER-2 status in breast cancer patients and analyze their relationship to clinical characteristics. Methods A total of 128 samples of breast cancer tissue were analyzed retrospectively. FISH was employed to detect the HER-2 gene amplification. And the FISH findings were compared with IHC test results by analyzing the concordance and correlation between two results. And their relationships to the clinical characteristics were analyzed. Results The overall coincidence rate of IHC and FISH was 90. 6% ( kappa = 0.405, P = 0. 000 ). And the discordance was mainly found in the IHC ( + + ) group. A positive correlation was found between the two results ( r =0. 655, P=0. 000). The ER (estrogen receptor) expression was negatively correlated with HER-2 gene amplification and the expression of Her-2 protein ( r = - 0. 300, P = 0. 001;r = - 0. 223, P = 0.011 ).There was a negative correlation between ER/PR status and HER-2 gene amplification (r = -0.213, P=0.016). The similar results were found in subgroup analysis. Tumor grade was negatively correlated with the expression of Her-2 protein ( r = - 0. 293, P = 0. 008 ), but not with HER-2 gene amplification ( P >0. 05). Conclusion IHC is a preferred method to detect the Her-2 status in breast cancer. The strong positive expression ( + + + ) of HER-2 protein tested by IHC is strongly consistent with HER-2 gene amplification by FISH. But HER-2 gene amplification should be further detected by FISH in patients with HER-2 positive expression ( + - + + ) in order to guide the clinical diagnosis and treatment. ER, ER/PR (progesterone receptor)status and tumor grade are correlated with HER-2 gene amplification and/or the expression of Her-2 protein. This study helps improve the accuracy of judging HER-2 gene amplification according to the clinical and pathological features such as ER status and the results of IHC.  相似文献   

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目的 比较荧光原位杂交(FISH)与免疫组织化学技术(IHC)在检测乳腺癌患者HER-2基因扩增和表达状态方面的应用.方法 采用FISH技术对66例IHC检测结果为HER-2基因过度表达(3+/2+)和低表达或无表达(1+/-)的乳腺癌石蜡切片进行HER-2基因扩增状态检测.结果 用FISH技术检测42例IHC结果显示HER-2基因过度表达(3+/2+)的标本,31例显示HER-2基因扩增,11例无扩增.检测24例IHC检测显示HER-2基因低度表达或无表达(1+/-)的标本,均未显示HER-2基因扩增.两组之间比较,Kappa系数为0.672,P<0.001,显示两项检测技术具有良好的一致性.另外.用FISH技术检测到部分病例显示17号染色体多体性,并且该多体性在HER-2高表达(3+/2+)的病例发生率显著高于低表达或无表达(1+/-)的病例(x~2=4.688,P=0.03).结论 FISH和ICH两项技术具有良好的一致性.IHC可作为HER-2基因扩增和表达状态检测的筛查手段.FISH技术可以作为检测HER-2基因扩增和17号染色体多体性的确诊手段.
Abstract:
Objective To evaluate the application of the immunohistochemistry (IHC) and the fluorescence in situ hybridization (FISH) in detecting the amplification and the expression of HER-2 gene in the breast cancer patients. Methods Sixty-six cases of paraffin-embeded breast cancer samples with overexpression, low or no expression of HER-2 gene as detected by IHC were analyzed for HER-2 gene amplification using FISH. Results Among the 42 samples with HER-2 gene overexpression (3+/2+) detected by IHC, 31 showed positive HER-2 gene amplification and 11 showed negative HER-2 gene amplification in FISH. In the 24 samples with low or no HER-2 gene expression (1+/-) detected by IHC, no HER-2 gene amplification was detected by FISH. The results of the two testing methods showed a good consistency with the kappa coefficient of 0.672 (P<0.001). We also found that the 17 chromosome polysomy in 42% of the samples and the incidence of 17 polysomy was significantly higher in the HER-2 gene overexpression (3+/2+) group than in low or no HER-2 gene expression (1+/-) group (x~2=4.688, P=0.03). Conclusion IHC can be used as a screening method for detecting HER-2 gene amplification, and FISH should be performed in cases of HER-2 gene overexpression (3+/2+) as detected by IHC.  相似文献   

10.
Background This study was designed to detect methylation of E-cadherin gene promoter and gene mutation of β-catenin in exon 3 and their expression of protein and mRNA in primary tumor and lymph node metastatic tumor of nasopharyngeal carcinoma (NPC), and investigate the mechanism of invasion and metastasis of neoplastic cells in NPC. Methods Fourty-two fresh biopsy samples were taken from untreated NPC patients at the Affiliated Hospital of Sun Yat-sen Medical College, Sun Yat-sen University, Guangzhou, China during the period of 1999 -2002. Among them 21 were taken from primary tumors and the other 21 from lymph node metastatic tumors. The gene promoter methylation of E-cadherin was detected by methylation-specific PCR (MSP). The mutation in exon 3 of β-catenin was detected by direct sequencing analysis. RT-PCR, Western blot and immunohistochemical staining were used to detect the mRNA and protein expression patterns in both primary and metastatic tumors of NPC. Results Down-regulated expression of E-cadherin in metastatic tumor was compared with that in primary tumor. Reduced expression of E-cadherin was found to be correlated with lymph node metastatic tumor of NPC ( P = 0. 004) ; but there was no obvious correlation between primary and metastatic tumors in the expression of β-catenin (P = 0. 698). The mRNA expression level of Ecadherin in metastatic tumors decreased significantly compared with that in primary tumors. However, little change was observed in the mRNA level of β-catenin in different tumor tissues. Only 4 samples (19. 1%) displayed gene promoter methylation of E-cadherin in primary tumor and 10 samples (47. 6%) showed methylated form of E-cadherin. The gene promoter methylation of E-cadherin was more common in metastatic tumor than in primary tumor of NPC ( P =0. 024). Only 2 (4. 76% ) of the 42 samples showed mutations in exon 3 of β-catenin at 41 (T41A, ACC→GCC) and codon 47(S47T, AGT→ACT). The cytoplasmic and nuclear expression of β-catenin in tumor was not found in any samples of NPC. Conclusions The results suggest that the downregulation of E-cadherin results from the gene promoter aberrant methylation of E-cadherin and that the methylation of E-cadherin plays an important role in invasion and metastasis of tumor cells in NPC. However, β-catenin mutation is an infrequent event in NPC, and β-catenin is not a critical factor influencing the invasion and metastasis of tumor cells in NPC.  相似文献   

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