Differential Expression of HER2 and SKP2 in Benign and Malignant Colorectal Lesions.

BACKGROUND: Colorectal cancer (CRC) is the fourth most common cancer worldwide. Both HER2 and SKP2 have a carcinogenic role in CRC making them attractive targets for tailored treatment. This work aims to correlate HER2 and SKP2 protein expression as well as HER2 gene amplification with clinicopathological parameters aiming at identifying potential candidates for targeted therapy. METHODS: This Study was conducted on 127 paraffin-embedded tissue samples of different colorectal lesions [controls, chronic colitis, ulcerative colitis (UC), hyperplastic polyps (HPs), adenomas and CRCs] to investigate HER2 and SKP2 expression by immunohistochemistry (IHC), Selected CRC cases [equivocal (2+) and positive (3+) by IHC] were further evaluated by ISH (CISH and SISH ) to assess HER2 gene amplification. RESULTS: Chronic colitis, UC, HPs and adenomas were HER2-negative. HER2 positivity (scores 2+ and 3+) was found only in15% of CRCs. Both SISH and CISH showed the same results with high concordance as 66.7% of equivocal and 100% of positive cases showed amplification of HER2 gene. SKP2 positivity was detected in 26.7% and 45% of adenomas and CRCs respectively, while other studied groups were negative. A significant correlation was noted between HER2 and SKP2 expression. CONCLUSION: A small percent of CRCs exhibited HER2 gene amplification, which would be potential candidates for anti HER2 therapy whereas IHC could be a primary screening test for patient selection. A potential carcinogenic role of SKP2 was suggested by the findings that SKP2 expression was undetectable in normal colonic mucosa but significantly increases from adenoma to carcinoma, hoping adenoma patients to get benefit from targeted therapy.
.

Attenuated isolated 3' signal: A highly challenging therapy relevant ALK FISH pattern in NSCLC.

OBJECTIVES: Targeted therapies in the management of patients with lung cancer provide significantly better outcome compared to chemotherapy. Detection of the anaplastic lymphoma kinase (ALK) gene rearrangement has great predictive value for treatment with small molecule tyrosine kinase inhibitor (crizotinib and alectinib commonly). Fluorescent in situ hybridisation (FISH) assay is a basic diagnostic test designed for detecting ALK gene rearrangements. Although being considered as gold standard method by IASLC's guideline, it is often regarded as difficult and error prone. Our aim was to examine a unique atypical ALK FISH pattern, revealed during a systematic large-scale monitoring, which carries the great risk of misinterpretation, hence may result in loss of patients eligible for targeted therapy. MATERIALS AND METHODS: Tissue and cytology samples from nearly one thousand patients with advanced stage non-small cell lung cancer (NSCLC, n = 996) were routinely examined by ALK FISH and immunohistochemistry (Ventana ALK-D5F3-CDx assay). Anchored Multiplex PCR based Next Generation Sequencing (AMP-NGS) was used to detect fusion gene transcripts in ambiguous cases. RESULTS: Fifty-nine (5,9%) of the cases were positive with ALK FISH test. Three cases showed atypical pattern with a significantly reduced sized red (3') signal and complete loss of green signals. Digital signal measurement confirmed this finding, showing consistent attenuation of 3' signals throughout the tumours. In all three cases AMP-NGS and ALK IHC verified the presence of a fusion gene and expressed oncoprotein, respectively. CONCLUSION: Approximately 5% of the 59 ALK positive cases exhibited atypical attenuated isolated 3' signal pattern. The immunohistochemistry and AMP-NGS examinations helped to clarify the presence of oncoprotein and the fusion gene, respectively. Our results emphasize the importance of extensive exploration of the genetic background of any unexpected FISH finding to avoid false diagnosis. This enables clinicians to indicate the adequate therapy with higher efficiency for patients suffering from NSCLC.

HER2 gene amplification in breast cancer by fluorescence in-situ hybridization and its clinicopathologic relationship / 临床与实验病理学杂志

Purpose To detect the HER2 gene amplification by fluorescence in-situ hybridization(FISH) and to explore its clinicopathologic relationship with the breast cancer.Methods A prospective study was conducted in 50 cases of breast invasive ductal carcinoma from Beijing Toren Hospital. Clinicopathologic data were summarized and FISH was performed in the paraffin-embedded sections for HER2 gene amplificayion using DNA probe, and immunohistochemical stain for ER, PR and HER2.Results The average age of the patients was 55.5 years. The pathologic grading showed that 11 cases were in grade Ⅰ, 30 cases in grade Ⅱ, and 9 cases in grade Ⅲ. The TNM staging showed that 13 cases were in stage Ⅰ,15 cases in ⅡA,13 cases in ⅡB,6 cases in ⅢA,2 cases in ⅢB,and 1 case in ⅢC.The median metastasis rate of lymph node was 6.91%.33 cases were positive for ER,and 32 cases positive for PR.For HER2 detection, 40 cases were positive by IHC and 33 positive by FISH. HER2 gene amplification by FISH was closely related with the expression of HER2 protein by immunohistochemistry, but not significantly related with pathologic grading, The TNM staging, median lymph node metastasis rate, ER and PR status (P>0.05). FISH test was positive in 3 cases of tumor embolus and 3 cases of multiple primary tumors. Conclusions FISH and immunohistochemistry for detecting HER2 have a good conformance, HER2 gene amplification may be related with tumor embolus and multiple primary tumors, but it can not be used as an indirect marker to predict the prognosis.

Clinicopathologic analysis of four cases of extranodal follicular dendritic cell sarcoma / 白血病·淋巴瘤

Objective To study the clinicopathoiogic features of four cases of extranodal follicular dendritic cell sarcoma(FDCS). Methods Four cases of FDCS were examined by histological and immunohistochemistry and in situ hybridization for Epstein-Barr virus (EBV)-encoded RNA, and the related literatures were reviewed. Results Microscopically, the neoplastic cells were spindle-shaped or ovoid with pale-stained cytoplasm, indistinct cell borders, granular chromatin, distinct small nucleoli. There were varied growth patterns in the tumour, such as fascicular, circular whorls and storiform, and abundant with intermixed small lymphocytes. There scattered multinucleated giant cells and perivascular cuffing phenomenon. There were rare mitoses. The neoplastic cells were positive for one or more of the follicular dendritic markers such as CD21, CD23 and CD35. It was variably positive for CD68, Vimentin, LCA and S-100. Staining for CD20, CD3 and CD1α were negative. Ki-67 labeling ranged from 10 %-20 %. The neoplastic cells were negative for Epstein-Barrvirus (EBV)-encoded RNA. Conclusion FDCS is a rare low-grade malignant tumor with varied growth pattern. The diagnosis should be confirmed by immunohistochemical staining. It is important to recognize its morphological characteristics to avoid confusion with other similar lesions, such as gastrointestinal stromal tumour, inflammatory pseudotumor, interdigitaing dendtritic cell sarcoma, malignant histiocytoma, lymphoepithelial carcinoma, diffuse large B-cell lymphoma, and malignant melanoma.

Assessment of HER2/neu status in breast cancer: a comparison of immunohistochemistry, fluorescence insitu hybridization and chromogenic insitu hybridization / 肿瘤研究与临床

Objective Through comparision of HER2/neu status detected by immunohistoehemistry (IHC),fluorescence insitu hybridization(FISH)and chromogenic insitu hybridization(CISH),to explore the clinical significance of FISH and CISH on detecting gene status of HER2.Methods Sixty-four paraffinembedded breast samples whose pathological types were infiltrating ductal carcinomas were tested by IHC for HER2 protein overexpression and CISH,FISH for HER2 gene status.The rate of chromosome 17 polysomy was also analyzed.Results Among the cases of HER2 protein overexpression(+++)group,the concordant ratios of IHC and CISH.FISH detecting HER2/neu status were 100%.Among the cases of HER2 protein overexpression(++)group,the concordant ratios were 95.83%and 91.67%.Among the cases of HER2 protein overexpression(+/-)group,the concordant ratios were also 100%.One case in which FISH and CISH were performed yielded the different results.so the general coincident rate was 98.41%.The incidence of chromosome 17 polysomy Was 45.16%,45.83%and 11.11%in three IHC(+++,++,+/-)groups,and the total incidence was 40.62%.ConclusionIHC is a preliminary screening method;HER2 gene status detected by CISH and FISH have high concordant ratio,CISH is a more convenience and viable method to detect HER2/neu status,when doubling interference of fichromosome 17 polysomy,we need to select FISH further.

Gene expression of interleukin-1 alpha in degenerative cervical discs and its significance / 中华物理医学与康复杂志

Objective To investigate the gene expression of interleukin-1 alpha (IL-1?) in degenerative cervical intervertebral discs and its significance. Methods The specimens of the cervical intervertebral discs were harvested from 12 patients with cervical disc herniation (CDH) and 16 patients with cervical spondylotic myelopathy (CSM) in surgical operations, and categorized according to the MRI disc degeneration classification. The specimens of 8 normal cervical intervertebral discs from fresh corpses were harvested as control group. The expression of IL-1? mRNA was detected by in situ hybridization histochemistry (ISHH) and the difference and change characteristics of IL-1? mRNA expression were compared among groups and each grades. Results The expression of IL-1? mRNA was obviously stronger in CDH and CSM groups than in the control group and stronger in discs of grades Ⅲ and Ⅳ than gradesⅠ and Ⅱ. Conclusion The gene expression of IL-1? was strengthened in degenerative cervical intevertebral discs and the more serious the degeneration, the stronger the gene expression. IL-1? may play an important role in the degeneration of cervical intervertebral discs.

Expression of Tissue Inhibitors of Metalloproteinases (TIMPs) in Hepatocellular Carcinoma

BACKGROUND: Matrix metalloproteinases (MMPs) have been implicated in the remodelling of extracellular matrix (ECM), including basement membrane. ECM remodelling is associated with pathological processes, including hepatic fibrosis, tumor invasion and metastasis. Tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 were known to inhibit MMP-9 and MMP-2, respectively. In the present study, we examined the expression of TIMP-1 and TIMP-2 in surgical specimen pairs of hepatocellular carcinoma and nontumoral liver and the correlation between their expression and clinicopathological characteristics. METHODS: The localization of both transcripts and protein of TIMP-1 and TIMP-2 was studied by using in situ hybridization and immunohistochemistry. RESULTS: TIMP-1 and TIMP-2 mRNA transcripts were found in tumor cells, hepatocyte, sinusoidal cells, endothelial cells and stromal cells. Signal intensity of TIMP-1 was stronger than that of TIMP-2. The results of immunohistochemical stainings were concordant with those obtained by in situ hybridization. Expression of TIMP-1 and TIMP-2 was observed in tumorous tissue, in nontumorous tissue and in the portions of the tumors adjacent to the capsules. However, a clear difference in TIMP-1 and TIMP-2 mRNA expression was not observed among the three tissue types. The intensity of TIMP-2 expression was generally weaker than that of TIMP-1, and the intensity of TIMP-1 and TIMP-2 mRNA expression did not correlate with variable clinicopathological characteristics. CONCLUSION: TIMPs was expressed in tumor cells and many cell types of the nontumoral liver. Further investigations for TIMPs' unknown functional role are needed.