OBJECTIVE: To investigate paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples. METHODS: p16 immunohistochemical staining was performed in 150 cases of chronic cervicitis, 126 cases of low-grade squamous intraepithelial lesions(LSIL), 96 cases of high-grade squamous intraepithelial lesions (HSIL) and 78 cases of cervical cancer from January 2014 to March 2018 in Zhongshan Boai Hospital. The results of p16 protein expression in paraffin sections with thickness of 2, 3, 4, 5 and 6 μm were compared using Logistic regression analysis. RESULTS: With the increase of slice thickness, the staining of p16 protein in nucleus gradually increased. The thickness of cervical slices in chronic cervicitis and cervical cancer samples had no significant effect on the positive rate of p16 protein(=7.817 and 1.332, both >0.05), while the thickness of slices in LSIL and HSIL samples had significant effect on the positive rate of p16 protein (=17.688 and 10.182, <0.05 or <0.01). The stable and reliable results were obtained when the slices were between 3 and 5 μm thick. CONCLUSIONS Paraffin sections with thickness of 3.0-5.0 μm are recommended for immnohistochemical staining of p16 protein in cervical tissue samples.
Biomarkers, Tumor ; genetics ; metabolism ; Cervical Intraepithelial Neoplasia ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Female ; Histocytological Preparation Techniques ; standards ; Humans ; Immunohistochemistry ; Paraffin ; Squamous Intraepithelial Lesions of the Cervix ; Uterine Cervical Neoplasms ; physiopathology

Biomarkers, Tumor ; metabolism ; Formaldehyde ; Humans ; Neoplasms ; metabolism ; Paraffin Embedding ; RNA, Messenger ; metabolism ; Tissue Fixation

No abstract available.
Adult ; Aged ; Base Sequence ; Bone Marrow/metabolism/pathology ; Female ; Formaldehyde/chemistry ; Gene Frequency ; Humans ; Male ; Middle Aged ; Multiple Myeloma/diagnosis/genetics ; Mutation ; Myeloid Differentiation Factor 88/chemistry/*genetics/metabolism ; Paraffin Embedding ; Sequence Analysis, DNA/*methods ; Waldenstrom Macroglobulinemia/diagnosis/genetics

OBJECTIVETo explore the feasibility of testing HER-2 expression and gene amplification in fine needle aspiration specimens of advanced breast cancers, and to benefit the patients receiving targeted drug therapy.

METHODSLiquid-based cytology specimens by fine needle aspiration of 49 breast cancer cases were used in this study. The expression of HER-2 protein was detected by immunocytochemistry (ICC) and the gene amplification was assessed by fluorescence in situ hybridization (FISH). All the 49 cases had overexpression of HER-2 protein marked as ++ or +++ in immunohistochemistry (IHC), and had corresponding FISH results in formalin-fixed, paraffin-embedded (FFPE) tissue samples.

RESULTSFNA samples in all the 49 cases were tested by FISH, and showed a complete agreement with the FISH results in the histological specimens (kappa = 1.0). Of the 49 cases, 33 had HER-2 gene amplification in FFPE samples. So do that in FNA samples. Both had an amplification rate of 67.3%. Among the 33 cases with HER-2 gene amplification, 26 had an ICC score of +++ (78.8%). The conformity rate was 78.8%. Of the 33 cases, 29 had an IHC score of +++ (87.9%). Its conformity rate was 87.9%. The difference between the ICC and IHC results was statistically not significant (P = 0.322). Among the 16 cases with negative gene amplification by both ICC and IHC, 15 cases showed HER-2 protein expression as 0/+, and another one case was not counted because there was not enough cells. Of the 16 cases, 15 had an IHC score of ++ and one of +++ . To take the FISH results as gold standard, ICC results had a high sensitivity (87.9%) and specificity (100.0%).

CONCLUSIONSFISH in FNA samples can be used in the clinic to test HER-2 gene amplification and overexpression in breast cancers, with a high sensitivity and specificity in ICC. Our data support the use of FISH and ICC analysis to determine HER-2 status on FNA specimens in patients with advanced breast cancer and recurrence or metastatic tumors. When ICC score is +++ , it indicates that there is a HER-2 gene amplification by FISH.

Adult ; Aged ; Biopsy, Fine-Needle ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; Female ; Gene Amplification ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Paraffin Embedding ; Receptor, ErbB-2 ; genetics ; metabolism ; Sensitivity and Specificity

OBJECTIVETo investigate the frequency of anaplastic lymphoma kinase (ALK) expression in non-small cell lung cancer (NSCLC) patients and its correlation with the clinicopathologic features.

METHODSALK immunohistochemistry and ALK fluorescent in situ hybridization (FISH) were performed on formalin-fixed, paraffin-embedded tissue in 100 cases of NSCLCs between 2011 and 2013. Relevant clinicopathologic data were collected and correlated with ALK expression.

RESULTSAll patients with immunohistochemical score of 3 (n = 12) were FISH-positive and all patients with score of 0 (n = 78) were FISH-negative. Among patients with immunohistochemical scores of 1 and 2, 2/3 and 6/7 were FISH-positive, respectively. The sensitivity and specificity of ALK immunohistochemistry with intensity score of 1 or more were 100% and 98%, respectively. Invasive mucinous adenocarcinoma, solid or acinar growth pattern, presence of mucous cells (signet-ring cells or goblet cells), extracellular mucus and lack of significant nuclear pleomorphism characterized ALK-rearranged cancer.

CONCLUSIONSALK-rearranged cancers possess specific histological features. Immunohistochemistry can be used as a routine test for screening ALK-positive cases in advanced NSCLC, and FISH testing should be used to confirm ALK translocation for patients with tumors showing staining for ALK by immunohistochemistry. All of these can help physicians identify patients who may benefit from targeted therapy.

Carcinoma, Non-Small-Cell Lung ; enzymology ; Female ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; enzymology ; Male ; Paraffin Embedding ; Receptor Protein-Tyrosine Kinases ; metabolism ; Sensitivity and Specificity

OBJECTIVETo evaluate the immunohistochemical detection of epidermal growth factor receptor(EGFR) mutations using two EGFR mutation-specific monoclonal antibodies: delE746-A750 and L858R.

METHODSA total of 175 paraffin-embedded lung adenocarcinoma tissue samples previously genotyped by directive DNA sequencing were subject to immunostaining using delE746-A750 and L858R antibodies.

RESULTSThere was no significant difference of mutation detection between DNA sequence analysis and delE746-A750 and/or L858R immunostaining (33.7% vs 30.9%, P > 0.05). The overall sensitivity, specificity, positive predictive value and negative predictive value of immunostaining using these two EGFR mutation-specific antibodies were 83.1%, 95.7%, 90.7% and 90.9%, respectively.

CONCLUSIONWith high sensitivity and good specificity, immunohistochemistry using EGFR mutation-specific monoclonal antibodies is an adequate, easy and cost-effective prescreening method to detect EGFR mutations using paraffin-embedded tissue specimens of lung adenocarcinomas.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Monoclonal ; DNA Mutational Analysis ; Female ; Gene Deletion ; Humans ; Immunohistochemistry ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Paraffin Embedding ; Point Mutation ; Receptor, Epidermal Growth Factor ; genetics ; immunology ; metabolism ; Sensitivity and Specificity

OBJECTIVETo examine the sensitivity and specificity of the two epidermal growth factor receptor hotspot mutation antibodies-delE746-A750 and L858R in lung adenocarcinoma, and to discuss the value of the two antibodies for detecting these two "classic mutations" by immunohistochemistry.

METHODSWith Q-scoring assessment and ROC curve analysis,immunohistochemistry was used to screen a panel of 309 paraffin-embedded non-small cell lung cancer(NSCLC) tumor samples, of which EGFR gene mutational status was previously documented by PCR-DNA sequencing.

RESULTSThree hundred and nine NSCLC tumor samples conclude 92 NSCLC cases with 19delE746-A750 and 110 with 21L858R. Using DelEGFR antibody to detect 19delE746-A750 demonstrated sensitivity and specificity of 84.8% (78/92) and 77.6% (83/107), respectively. Choosing the best cut-off value of 85.00, it achieved the maximum area under the curve (AUC:0.885). L858R antibody to detect 21L858R showed sensitivity and specificity of 94.5% (104/110) and 62.5% (55/88), respectively. Choosing the best cut-off value of 97.50, it achieved the maximum area under the curve (AUC:0.924).

CONCLUSIONBy using Q-scoring assessment and ROC curve analysis, the two antibodies (delEGFR and L858R) can been used to identify EGFR gene mutations present in lung adenocarcinomas.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Aged ; Antibodies, Monoclonal ; Area Under Curve ; Exons ; Female ; Gene Deletion ; Humans ; Immunohistochemistry ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Paraffin Embedding ; Point Mutation ; Receptor, Epidermal Growth Factor ; genetics ; Sensitivity and Specificity

OBJECTIVETo use array-based comparative genomic hybridization (aCGH) technology to study the molecular cytogenetic abnormalities of anaplastic large cell lymphoma (ALCL) at genome level.

METHODSALK protein expression and molecular genetic abnormalities were detected by immunohistochemistry and fluorescence in situ hybridization, respectively, in 25 cases of ALCL. Any chromosomal gains/losses were detected by aCGH and correlated with ALK status.

RESULTSaCGH showed that chromosomal alterations in all 25 ALCL cases, and the frequency of chromosomal gains was higher than that of the losses. Chromosomal gains at 5p13.2, 3q21.1, 2q21.3, 3p25.1, 14q32.33, and 17q21.2 regions were detected in more than 50% of the ALCL cases; gains at 4q27, 6p22.1, 20p11.21, 2q22.3, 4q35.1, 1p36.22, 8p23.1, 8p12, 11q14.1, 12q13.13, and 19p13.3 regions were detected in 30%-50% of the ALCL cases; chromosomal losses at 3q26.1 and 3q26.31 regions were detected in 36.0% (9/25) and 24.0% (6/25) of the ALCL cases, respectively. Chromosomal gains at 2q21.3, 6p22.1 and 3p25.1 regions showed significant differences between ALK (+) and ALK (-) ALCL groups (P < 0.05).

CONCLUSIONSaCGH demonstrates complex molecular genetic variations in all ALCL cases. Gains at 2q21.3, 6p22.1 and 3p25.1 regions are significantly different between ALK (+) and ALK (-) ALCL groups, suggesting that the pathogenesis of ALK (+) and ALK (-) ALCL may involve different signaling pathway.

Adolescent ; Chromosome Aberrations ; Comparative Genomic Hybridization ; methods ; Female ; Gene Expression Profiling ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Large-Cell, Anaplastic ; enzymology ; genetics ; Male ; Paraffin Embedding ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism

OBJECTIVETo investigate the standard protocol for anaplastic lymphoma kinase (ALK) fusion gene assessment by fluorescent in situ hybridization (FISH) in non-small cell lung cancer (NSCLC).

METHODSTissue specimens of NSCLC cases were retrospectively collected from Jan. 2011 to July 2012. ALK fusion gene was examined by FISH using break-apart ALK gene probes (Vysis company). The identification of ALK fusion gene was determined by fluorescent signals under a fluorescence microscope.

RESULTSOne hundred and forty-six eligible NSCLC tumor specimens were tested for ALK fusion gene by FISH. The specimens included 110 cases (75.4%) of surgically-removed tissues, 11 cases (7.5%) of biopsy, 19 cases (13.0%) of lymph node and 6 cases (4.1%) of other metastatic tissues. The positivity of ALK fusion gene was 8.9% (13/146).

CONCLUSIONSThe assessment of ALK fusing gene by FISH using standard protocol for formalin-fixed, paraffin-embedded (FFPE) tissue is feasible. The protocol can used to test in surgically-removed tissues, biopsies, metastatic lymph nodes and other metastastic specimens.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; surgery ; Female ; Gene Fusion ; Humans ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; genetics ; metabolism ; surgery ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; metabolism ; Paraffin Embedding ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism ; Retrospective Studies ; Young Adult

Carcinoma, Hepatocellular ; pathology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; MicroRNAs ; metabolism ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Staging ; Paraffin Embedding ; Real-Time Polymerase Chain Reaction ; Up-Regulation