Analysis of HER2 gene amplification using Differential PCR in breast cancer patients of Isfahan Province
Background: Amplification of HER2 is seen in 20-30% of breast cancer cases. Measurement of HER2 gene amplification appears to be of vital importance in planning the treatment schedule for patients with breast carcinoma. The aim of our study was to evaluate HER2 amplification status in malignant and...
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Mazandaran University of Medical Sciences and Health Services
2014-10-01
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doaj-6493cd1ca29946b9b4d6076c455eabb12020-11-25T03:42:22ZengMazandaran University of Medical Sciences and Health ServicesResearch in Molecular Medicine2322-13482322-133X2014-10-01241217Analysis of HER2 gene amplification using Differential PCR in breast cancer patients of Isfahan ProvinceZohreh Hojati0Zeinab Hallajian1Abolghasem Esmaeili2Majid Motovali-Bashi3Hosein Tabatabaeian4 Genetics Division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran. PO Box: 81746- 73441 Genetics Division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran. PO Box: 81746- 73441 Genetics Division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran. PO Box: 81746- 73441 Genetics Division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran. PO Box: 81746- 73441 Genetics Division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran. PO Box: 81746- 73441 Background: Amplification of HER2 is seen in 20-30% of breast cancer cases. Measurement of HER2 gene amplification appears to be of vital importance in planning the treatment schedule for patients with breast carcinoma. The aim of our study was to evaluate HER2 amplification status in malignant and benign breast tumors by differential PCR (dPCR). Materials and Methods: The genomic DNA was extracted using the phenol/chloroform extraction procedure from 76 different breast tissues. Differential PCR was performed using the DNA samples isolated from fresh and paraffin- embedded breast cancer tissues. The relative copy number ratio of target gene (HER2) to control gene ( INF-γ ) was measured. dPCR products were then separated by electrophoresis using 2% agarose gel. The intensity of HER2 and INFγ bands were determined for each sample by ImageJ software. Results: According to the ratio between the band intensity of HER2 to INFγ in tumour and also normal samples, 7% and 26% rates of HER2 amplification were observed in benign and malignant samples respectively. The ratio showed a 2-5 fold increase in HER2 gene copy number for tissues with HER2 amplification whereas, a one-fold increase was found in other samples. Conclusion: Differential PCR provides a relatively rapid and inexpensive technique to assess the HER2 gene amplification, especially alongside immunohistochemistry as a routine assessing method .http://rmm.mazums.ac.ir/browse.php?a_code=A-10-615-8&slc_lang=en&sid=1HER2 amplification breast cancer differential PCR |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Zohreh Hojati Zeinab Hallajian Abolghasem Esmaeili Majid Motovali-Bashi Hosein Tabatabaeian |
spellingShingle |
Zohreh Hojati Zeinab Hallajian Abolghasem Esmaeili Majid Motovali-Bashi Hosein Tabatabaeian Analysis of HER2 gene amplification using Differential PCR in breast cancer patients of Isfahan Province Research in Molecular Medicine HER2 amplification breast cancer differential PCR |
author_facet |
Zohreh Hojati Zeinab Hallajian Abolghasem Esmaeili Majid Motovali-Bashi Hosein Tabatabaeian |
author_sort |
Zohreh Hojati |
title |
Analysis of HER2 gene amplification using Differential PCR in breast cancer patients of Isfahan Province |
title_short |
Analysis of HER2 gene amplification using Differential PCR in breast cancer patients of Isfahan Province |
title_full |
Analysis of HER2 gene amplification using Differential PCR in breast cancer patients of Isfahan Province |
title_fullStr |
Analysis of HER2 gene amplification using Differential PCR in breast cancer patients of Isfahan Province |
title_full_unstemmed |
Analysis of HER2 gene amplification using Differential PCR in breast cancer patients of Isfahan Province |
title_sort |
analysis of her2 gene amplification using differential pcr in breast cancer patients of isfahan province |
publisher |
Mazandaran University of Medical Sciences and Health Services |
series |
Research in Molecular Medicine |
issn |
2322-1348 2322-133X |
publishDate |
2014-10-01 |
description |
Background: Amplification of HER2 is seen in 20-30% of breast cancer cases. Measurement of HER2 gene amplification appears to be of vital importance in planning the treatment schedule for patients with breast carcinoma. The aim of our study was to evaluate HER2 amplification status in malignant and benign breast tumors by differential PCR (dPCR). Materials and Methods: The genomic DNA was extracted using the phenol/chloroform extraction procedure from 76 different breast tissues. Differential PCR was performed using the DNA samples isolated from fresh and paraffin- embedded breast cancer tissues. The relative copy number ratio of target gene (HER2) to control gene ( INF-γ ) was measured. dPCR products were then separated by electrophoresis using 2% agarose gel. The intensity of HER2 and INFγ bands were determined for each sample by ImageJ software. Results: According to the ratio between the band intensity of HER2 to INFγ in tumour and also normal samples, 7% and 26% rates of HER2 amplification were observed in benign and malignant samples respectively. The ratio showed a 2-5 fold increase in HER2 gene copy number for tissues with HER2 amplification whereas, a one-fold increase was found in other samples. Conclusion: Differential PCR provides a relatively rapid and inexpensive technique to assess the HER2 gene amplification, especially alongside immunohistochemistry as a routine assessing method . |
topic |
HER2 amplification breast cancer differential PCR |
url |
http://rmm.mazums.ac.ir/browse.php?a_code=A-10-615-8&slc_lang=en&sid=1 |
work_keys_str_mv |
AT zohrehhojati analysisofher2geneamplificationusingdifferentialpcrinbreastcancerpatientsofisfahanprovince AT zeinabhallajian analysisofher2geneamplificationusingdifferentialpcrinbreastcancerpatientsofisfahanprovince AT abolghasemesmaeili analysisofher2geneamplificationusingdifferentialpcrinbreastcancerpatientsofisfahanprovince AT majidmotovalibashi analysisofher2geneamplificationusingdifferentialpcrinbreastcancerpatientsofisfahanprovince AT hoseintabatabaeian analysisofher2geneamplificationusingdifferentialpcrinbreastcancerpatientsofisfahanprovince |
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