Immunohistochemical and molecular study in women with breast cancer

العناوين الأخرى

دراسة مناعية نسيجية و جزيئية للنساء العراقيات المصابات بسرطان الثدي

مقدم أطروحة جامعية

al-Budayri, Iman Hatif Muhsin

مشرف أطروحة جامعية

al-Azzawi, Hasan Fayyd Samir
al-Qazzaz, Abd al-Karim Abd al-Razzaq Abd al-Wahhab

الجامعة

جامعة بغداد

الكلية

كلية العلوم

القسم الأكاديمي

قسم التقانة الإحيائية

دولة الجامعة

العراق

الدرجة العلمية

ماجستير

تاريخ الدرجة العلمية

2014

الملخص الإنجليزي

This study was done in the immunohistchimestry laboratory of ALSADER hospital/ AL-NAJAF AL ASHRAF and International Institute of early detection of cancer /Medical City /Baghdad.

Confirmed 50 cases of breast carcinoma (50 female cases with breast cancer were (43infiltrating ductal carcinoma IDC), 2 infiltrating lobular carcinoma (ILC), and 5 other types carcinoma), included in this study were reviewed in the period between January 2010 and April 2013 against a group of 20 cases with benign breast lesions was included for comparison.

Immunohistochemistry (IHC) were used to evaluate the expression of Her2/neu, Estrogen(ER), and Progesterone(PR) receptor test and to correlate immunohistochemical findings with prognostic parameters for breast carcinoma such as patients' age, tumor size, histological type, histological grade .

All samples from patients under study were included paraffin embedded formalin fixed blocks tissue.

Results observed that estrogen positive receptors were in 48% (24/50) of the cases and progesterone positive receptors in 46 % (23/50) of the cases, and these data revealed that presence of hormone receptor expression in the majority and cancer was considered hormone receptor positive.

The present results on Iraqi women patients revealed that a high age frequency of cancer occurred between 41-50 years of age (46%), and more than 24% of the patients have family history either it is the first or second degree.

The histopathology diagnosis showed that a high percentage in Iraqi cases with infiltrated ductal carcinoma represented (86%), while the invasive lobular carcinoma represented (4%) and the mixed carcinoma was represented (10%).

Results revealed that 12 % of patients were in grade 1, 20 % were in grade 2 and 68 % were in grade 3, so most of our patients were in grade 2 and 3.

Clinicopathological assessments for ER and PR revealed that there was no statistically significant association between expression and the age, family history, tumor types or stages (P>0.05).

On the other hand the expression of Her-2/neu did not show any statistical significant difference with the age, tumor grade or type except there was a significant correlation between her2/neu expression and family history (P<0.01).

About 56% of positive ER had a strong positive stain while 42% of PR had strong positive stain.

The results of immunohistochemical analysis for all breast cancer subtypes according to molecular subtype classification showed that luminal A was 54%, Luminal B 4%, Her2/neu subtype 10% and Basel like 32%.

Her2/neu over-expression was observed in 14% of Iraqi breast carcinoma affecting female patients.

This group presents apparently an aggressive form of breast carcinoma with high histological grade and negative ER.

Four different DNA extraction protocols (Phenol/chloroform, Genaid, Qiagen, and Promega extractions kits) were carried out to evaluate the yield and quality of DNA from formalin fixed paraffin embedded tissue samples(FFPE).

The results demonstrated the successful use and optimization of a rapid, reliable and effective protocol for DNA amplification directly from paraffin-embedded tissue eliminating the deparaffinization and DNA isolation steps with maximum yield in the range of was 160 - 293μg obtained with Promega DNA extraction kit , while all of the other kits produce smear DNA in most FFPE cases.

For analysis of HER2/neu gene amplification in FFPE breast cancer cases a standard PCR was used by using forward and reverse primers were performed and PCR amplified products were analyzed by 2% agarose gel electrophoresis and ethidium bromide staining against benign tissue.

HER2/neu amplified fragment band displayed a strong band in one case and estimated to have a size of 126 bp.

A HER2/neu DNA band was extracted from lane and purified using a gel extraction kit and sequenced using the ABI DNA sequencing kit.

Samples were loaded into the ABI 310 Genetic Analyzer and the results are shown.

BLAST analysis of the DNA sequence was carried out in the National Center for Biotechnology Information’s (NCBI) Genbank and found that the sequence corresponds to the HER2/neu gene fragment on exon 3.

التخصصات الرئيسية

الأحياء

الموضوعات

• السرطان(مرض)
• العراق
• الثدي
• الجوانب الوراثية

عدد الصفحات

155

قائمة المحتويات

• APA
• MLA
• AMA

لغة النص

الإنجليزية

نوع البيانات

رسائل جامعية

رقم السجل

BIM-601129