Quantification of Her-2/Neu gene in breast cancer patients using real time-polymerase chain reaction (Q-PCR) and correlation with immunohistochemistry findings

Nor Azian Abdul Murad, Zuraini Abdul Razak, Rosniza Muhammmad Hussain, Sharifah Noor Akmal Syed Hussain, Clarence Ko Ching Huat, Siti Aishah Che Md Ali, Norlia Abdullah, Rohaizak Muhammad, Naqiyah Ibrahim, A. Rahman A. Jamal

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Abstract

Background: HER-2/neu is a proto-oncogene that encodes a transmembrane tyrosine kinase growth factor which is crucial for stimulating growth and cellular motility. Overexpression of HER-2/neu is observed in 10-35% of human breast cancers and is associated with pathogenesis, prognosis as well as response to therapy. Given the imperative role of HER-2/neu overexpression in breast cancer, it is important to determine the magnitude of amplification which may facilitate a better prognosis as well as personalized therapy in affected patients. In this study, we determined HER-2/neu protein expression by immunohistochemistry (IHC) concurrently with HER-2/neu DNA amplification by quantitative real time-polymerase chain reaction (Q-PCR). Materials and Methods: A total of 53 paired tissue samples from breast cancer patients were frozen-sectioned to characterize the tumour and normal tissues. Only tissues with 80% tumour cells were used in this study. For confirmation, Q-PCR was used to determine the HER-2/neu DNA amplification. Results: We found 20/53 (37.7%) of the tumour tissues to be positive for HER-2/neu protein overexpression using IHC. Out of these twenty, only 9/53 (17%) cases were in agreement with the Q-PCR results. The concordance rate between IHC and Q-PCR was 79.3%. Approximately 20.7% of positive IHC cases showed no HER-2/neu gene amplification using Q-PCR. Conclusion: In conclusion, IHC can be used as an initial screening method for detection of the HER-2/neu protein overexpression. Techniques such as Q-PCR should be employed to verify the IHC results for uncertain cases as well as determination of HER-2/neu gene amplification.

Original languageEnglish
Pages (from-to)1655-1659
Number of pages5
JournalAsian Pacific Journal of Cancer Prevention
Volume14
Issue number3
DOIs
Publication statusPublished - 2013

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erbB-2 Genes
Real-Time Polymerase Chain Reaction
Immunohistochemistry
Breast Neoplasms
Gene Amplification
Neoplasms
Proteins
Proto-Oncogenes
DNA
Protein-Tyrosine Kinases
Intercellular Signaling Peptides and Proteins
Therapeutics
Growth

Keywords

  • 2/neu gene quantification
  • Breast cancer
  • Her
  • Polymerase chain reaction (Q-PCR)
  • Real time

ASJC Scopus subject areas

  • Oncology
  • Cancer Research
  • Public Health, Environmental and Occupational Health
  • Epidemiology

Cite this

Quantification of Her-2/Neu gene in breast cancer patients using real time-polymerase chain reaction (Q-PCR) and correlation with immunohistochemistry findings. / Abdul Murad, Nor Azian; Abdul Razak, Zuraini; Hussain, Rosniza Muhammmad; Syed Hussain, Sharifah Noor Akmal; Ching Huat, Clarence Ko; Che Md Ali, Siti Aishah; Abdullah, Norlia; Muhammad, Rohaizak; Ibrahim, Naqiyah; A. Jamal, A. Rahman.

In: Asian Pacific Journal of Cancer Prevention, Vol. 14, No. 3, 2013, p. 1655-1659.

Research output: Contribution to journalArticle

Abdul Murad, Nor Azian ; Abdul Razak, Zuraini ; Hussain, Rosniza Muhammmad ; Syed Hussain, Sharifah Noor Akmal ; Ching Huat, Clarence Ko ; Che Md Ali, Siti Aishah ; Abdullah, Norlia ; Muhammad, Rohaizak ; Ibrahim, Naqiyah ; A. Jamal, A. Rahman. / Quantification of Her-2/Neu gene in breast cancer patients using real time-polymerase chain reaction (Q-PCR) and correlation with immunohistochemistry findings. In: Asian Pacific Journal of Cancer Prevention. 2013 ; Vol. 14, No. 3. pp. 1655-1659.
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abstract = "Background: HER-2/neu is a proto-oncogene that encodes a transmembrane tyrosine kinase growth factor which is crucial for stimulating growth and cellular motility. Overexpression of HER-2/neu is observed in 10-35{\%} of human breast cancers and is associated with pathogenesis, prognosis as well as response to therapy. Given the imperative role of HER-2/neu overexpression in breast cancer, it is important to determine the magnitude of amplification which may facilitate a better prognosis as well as personalized therapy in affected patients. In this study, we determined HER-2/neu protein expression by immunohistochemistry (IHC) concurrently with HER-2/neu DNA amplification by quantitative real time-polymerase chain reaction (Q-PCR). Materials and Methods: A total of 53 paired tissue samples from breast cancer patients were frozen-sectioned to characterize the tumour and normal tissues. Only tissues with 80{\%} tumour cells were used in this study. For confirmation, Q-PCR was used to determine the HER-2/neu DNA amplification. Results: We found 20/53 (37.7{\%}) of the tumour tissues to be positive for HER-2/neu protein overexpression using IHC. Out of these twenty, only 9/53 (17{\%}) cases were in agreement with the Q-PCR results. The concordance rate between IHC and Q-PCR was 79.3{\%}. Approximately 20.7{\%} of positive IHC cases showed no HER-2/neu gene amplification using Q-PCR. Conclusion: In conclusion, IHC can be used as an initial screening method for detection of the HER-2/neu protein overexpression. Techniques such as Q-PCR should be employed to verify the IHC results for uncertain cases as well as determination of HER-2/neu gene amplification.",
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T1 - Quantification of Her-2/Neu gene in breast cancer patients using real time-polymerase chain reaction (Q-PCR) and correlation with immunohistochemistry findings

AU - Abdul Murad, Nor Azian

AU - Abdul Razak, Zuraini

AU - Hussain, Rosniza Muhammmad

AU - Syed Hussain, Sharifah Noor Akmal

AU - Ching Huat, Clarence Ko

AU - Che Md Ali, Siti Aishah

AU - Abdullah, Norlia

AU - Muhammad, Rohaizak

AU - Ibrahim, Naqiyah

AU - A. Jamal, A. Rahman

PY - 2013

Y1 - 2013

N2 - Background: HER-2/neu is a proto-oncogene that encodes a transmembrane tyrosine kinase growth factor which is crucial for stimulating growth and cellular motility. Overexpression of HER-2/neu is observed in 10-35% of human breast cancers and is associated with pathogenesis, prognosis as well as response to therapy. Given the imperative role of HER-2/neu overexpression in breast cancer, it is important to determine the magnitude of amplification which may facilitate a better prognosis as well as personalized therapy in affected patients. In this study, we determined HER-2/neu protein expression by immunohistochemistry (IHC) concurrently with HER-2/neu DNA amplification by quantitative real time-polymerase chain reaction (Q-PCR). Materials and Methods: A total of 53 paired tissue samples from breast cancer patients were frozen-sectioned to characterize the tumour and normal tissues. Only tissues with 80% tumour cells were used in this study. For confirmation, Q-PCR was used to determine the HER-2/neu DNA amplification. Results: We found 20/53 (37.7%) of the tumour tissues to be positive for HER-2/neu protein overexpression using IHC. Out of these twenty, only 9/53 (17%) cases were in agreement with the Q-PCR results. The concordance rate between IHC and Q-PCR was 79.3%. Approximately 20.7% of positive IHC cases showed no HER-2/neu gene amplification using Q-PCR. Conclusion: In conclusion, IHC can be used as an initial screening method for detection of the HER-2/neu protein overexpression. Techniques such as Q-PCR should be employed to verify the IHC results for uncertain cases as well as determination of HER-2/neu gene amplification.

AB - Background: HER-2/neu is a proto-oncogene that encodes a transmembrane tyrosine kinase growth factor which is crucial for stimulating growth and cellular motility. Overexpression of HER-2/neu is observed in 10-35% of human breast cancers and is associated with pathogenesis, prognosis as well as response to therapy. Given the imperative role of HER-2/neu overexpression in breast cancer, it is important to determine the magnitude of amplification which may facilitate a better prognosis as well as personalized therapy in affected patients. In this study, we determined HER-2/neu protein expression by immunohistochemistry (IHC) concurrently with HER-2/neu DNA amplification by quantitative real time-polymerase chain reaction (Q-PCR). Materials and Methods: A total of 53 paired tissue samples from breast cancer patients were frozen-sectioned to characterize the tumour and normal tissues. Only tissues with 80% tumour cells were used in this study. For confirmation, Q-PCR was used to determine the HER-2/neu DNA amplification. Results: We found 20/53 (37.7%) of the tumour tissues to be positive for HER-2/neu protein overexpression using IHC. Out of these twenty, only 9/53 (17%) cases were in agreement with the Q-PCR results. The concordance rate between IHC and Q-PCR was 79.3%. Approximately 20.7% of positive IHC cases showed no HER-2/neu gene amplification using Q-PCR. Conclusion: In conclusion, IHC can be used as an initial screening method for detection of the HER-2/neu protein overexpression. Techniques such as Q-PCR should be employed to verify the IHC results for uncertain cases as well as determination of HER-2/neu gene amplification.

KW - 2/neu gene quantification

KW - Breast cancer

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