Quantitative PCR analysis of c-erb B-2 (HER2/neu) gene amplification and comparison with p185HER2/neu protein expression in breast cancer drill biopsies

In breast cancer amplification of the HER-2/neu oncogene and over-expression of the protein product is associated with poor prognosis, predicts response to some chemotherapeutic regimens and is the target for Herceptin treatment. To date there are several methods to assess the amplification/overexpression of HER-2/neu with each having advantages and disadvantages. We have studied amplification and over-expression of HER-2/neu in 250 consecutive cases of breast cancer (220 invasive and 30 in situ carcinomas) presenting to the Department of Pathology at Women's College Campus of Sunnybrook and Women's College Health Sciences Center. Thirty percent of the invasive carcinomas were node positive. HER-2/neu protein overexpression was assessed by immunohistochemistry (IH) using antibody CB11 and amplification of the gene by differential PCR. The percentage of tumor cells showing CB11 staining was determined and the most significant cut off point for positivity was >10% moderate or strong complete membranous staining. The gene was considered amplified if the density score of the product was >2. There was 94% concordance between the two methods (P value .0001). Both methods were positive in 16% of cases and negative in 78% of cases. Discrepant cases were examined by FISH which confirmed the IH results in 9/11 invasive carcinomas. These results show that there is excellent concordance between IH and PCR. However, immunohistochemistry is easier to perform and cheaper than PCR and could be used in routine assessment of HER-2/neu in breast cancer patients.

show abstract

“…The sensitivity was 91% and the specificity was 96%. Similar results correlating PCR and IH were obtained by P. Decremoux (23) and O'Malley (22).…”

Section: Discussionsupporting

confidence: 88%

Defining a Test for HER-2/neu Evaluation in Breast Cancer in the Diagnostic Setting

et al. 2001

View full text Add to dashboard Cite

show abstract

“…In earlier studies, the status of ERBB2 was evaluated by Southern, Northern, or Western blot, and, more recently, by polymerase chain reaction. Results of these methods may be affected by the dilution of tumor material by normal cells (2,21). These drawbacks have prompted the development of in situ techniques, FISH and IHC, which enable the recognition of the marker of interest in tumor cells only.…”

Section: Discussionmentioning

confidence: 99%

“…The range of positivity found in the various studies seems to be rather large: 5 to 55% for the amplification, and 10 to 55% for the overexpression (6). Indeed, results of molecular methods may be affected by the dilution of tumor material by normal cells and by the determination of the threshold defining the existence of an amplification or an overexpression (2,7,21). Recent technical developments have made possible the detection of the ERBB2 gene in tissue sections, touch preparations, and fine-needle aspirates using fluorescent in situ hybridization (FISH) (16,(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32).…”

mentioning

confidence: 98%

Strong Correlation between Results of Fluorescent In Situ Hybridization and Immunohistochemistry for the Assessment of the ERBB2 (HER-2/neu) Gene Status in Breast Carcinoma

et al. 2000

Self Cite

View full text Add to dashboard Cite

show abstract

“…Gene amplification, as analyzed by quantitative PCR, often correlates with protein expression determined by semiquantitative immunohistochemistry (26), but relationships with gene transcription have not been established, because of the lack of an accurate method for quantitating transcripts. Real-time quantitative RT-PCR, which has recently become available, can be used to determine relative amounts of mRNAs in cells and tissues, with a high sensitivity and accuracy (27).…”

Section: Discussionmentioning

confidence: 99%

Sodium Iodide Symporter and Pendrin Expression in Human Thyroid Tissues

Mian

Lacroix²,

Alzieu³

et al. 2001

Thyroid

View full text Add to dashboard Cite

Thyroid cells synthesize thyroid hormones through a multistep process during which iodide is transported through the basolateral and the apical membranes of thyrocytes. Two genes that participate in these transports and the corresponding proteins, namely sodium iodide symporter (NIS) and pendrin, the product of the Pendred syndrome gene, have recently been characterized. We studied NIS and pendrin expression at the mRNA and protein levels by a quantitative reverse transcription-polymerase chain reaction (RT-PCR) method and by single and double immunostaining in normal and pathological human thyroid tissues. In normal tissue, NIS and pendrin were detected in about 20% and 40%-60% of thyrocytes, respectively. The number of NIS- and pendrin-positive cells was much higher in hyperfunctioning tissue from Graves' disease or toxic adenoma. In hypofunctioning adenomas and carcinomas, the number of NIS- and pendrin-positive cells was low or nonexistent. Three types of follicular cells were observed in positive tissues: NIS-negative/pendrin-negative cells, NIS-positive/pendrin-positive cells, and NIS-negative/pendrin-positive cells. The first two types of cells appear to be resting and active cells, respectively, but the functional status of NIS-negative/pendrin-positive thyrocytes remains to be determined.

show abstract

Quantitative PCR analysis of c-erb B-2 (HER2/neu) gene amplification and comparison with p185HER2/neu protein expression in breast cancer drill biopsies

Cited by 26 publications

References 21 publications

Defining a Test for HER-2/neu Evaluation in Breast Cancer in the Diagnostic Setting

Defining a Test for HER-2/neu Evaluation in Breast Cancer in the Diagnostic Setting

Strong Correlation between Results of Fluorescent In Situ Hybridization and Immunohistochemistry for the Assessment of the ERBB2 (HER-2/neu) Gene Status in Breast Carcinoma

Sodium Iodide Symporter and Pendrin Expression in Human Thyroid Tissues

Contact Info

Product

Resources

About