Expression of BRCA1 mRNA in cancerous and non-cancerous
breast tissue of Bangladeshi females attending a tertiary care hospital
Authors
- Latifa Nishat
Department of Anatomy, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh https://orcid.org/0000-0003-1393-6927 - Sufi Hannan Zulfiqar RahmanDepartment of Immunology and Molecular Biology, National Institute of Cancer Research and Hospital, Dhaka, Bangladesh https://orcid.org/0000-0001-6202-7004
- Farida ArjumanDepartment of Histopathology, National Institute of Cancer Research and Hospital, Dhaka, Bangladesh
https://orcid.org/0009-0001-7010-7423 - Zinnat Ara YesminDepartment of Anatomy, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh
https://orcid.org/0000-0002-2163-6126 - Lutfun NaharDepartment of Anatomy, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh https://orcid.org/0009-0008-8723-7081
- Umma Habiba LaboniDepartment of Anatomy, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh https://orcid.org/0009-0009-5110-214X
- Samira Sultana AmeeDepartment of Anatomy, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh
- Farzana AfrozeDepartment of Anatomy, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh https://orcid.org/0009-0009-5936-5614
DOI:
https://doi.org/10.3329/bsmmuj.v18i1.76754Keywords
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Ethical approval
Ethics Committee of National Institute of Cancer Research and Hospital (No. NICRH/Ethics/2021/345
dated 29 Nov 2021).
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Published by Bangabandhu Sheikh
Mujib Medical University
Methods: The cross-sectional study was conducted on 50 breast cancer and 19 non-cancerous females. RNA was extracted from formalin-fixed paraffin-embedded breast tissue. Real-time RT-PCR was done for measuring BRCA1 mRNA. BRCA1 mRNA expressions in cancerous and non-cancerous breast tissue were compared and analysed.
Results: BRCA1 mRNA expression was reduced or absent in most of the cancerous and non-cancerous (consisting of fibroadenoma, fibrocystic disease, ductal hyperplasia and normal breast tissue) breast tissue. Expression of BRCA1 in breast cancer and fibroadenoma was almost similar statistically. All cancers were invasive ductal carcinoma and of grade II, and most of them were sporadic (86%). BRCA1 expression was not associated with reproductive or cancer-related characteristics except consanguinity of marriage. The non-cancerous females were younger than the cancer patients (33.5 versus 43 years, respectively).
Conclusion: The study suggests the necessity of bringing fibroadenoma patients, in addition to breast cancer patients, into the screening programme and analysing the molecular profile because their BRCA1 expression is similar. The study suggests the necessity of bringing fibroadenoma patients, in addition to breast cancer patients, into the screening programme and analysing the molecular profile because their BRCA1 expression is similar.
Mutations in the BRCA1 gene are found to be associated with breast cancer, and the frequency of these genetic mutations varies among ethnic groups and countries [2]. Methylation in BRCA1 promoter region, low expression, and copy number deletions also cause deficiency in BRCA1 and produce similar phenotypic features of tumours due to BRCA1 mutations [4, 5]. Dysfunction of this gene is associated with ovarian, prostatic, gastric and pancreatic cancers in addition to breast cancer [6, 7].
BRCA1 is a DNA repair gene. Its mRNA is expressed at the late G1/early S phase of the cell cycle before DNA synthesis. Expression of the BRCA1 protein closely follows its mRNA. Deficiency in this gene produces low expression of BRCA1 mRNA. Recent studies have shown that BRCA1-related breast cancers have clinico-pathological features that are usually associated with a poor prognosis, such as high-grade oestrogen receptor (ER) and progesterone receptor (PR) negative status, and over expression of the Her-2-Neu [8]. Studies also found that the expression of BRCA1 mRNA influences the effectiveness of chemotherapy and helps in the prediction of survival of the patients [9].
Other than breast cancer, BRCA1 gene mutations are found to be present in some benign non-cancerous breast lesions. Among the breast pathologies, most of the breast lesions are benign. A survey of literature from 1985 to 2019 found around 50% of clinical breast lesions were mastalgia and fibrocystic disease, and 25% were fibroadenomas [10]. Benign breast diseases are evidenced as a risk factor for breast cancer, and the risk is higher with BRCA1 mutations [11].
Although high-income countries have achieved significant progress toward curing women with breast cancer, Bangladesh, like other low-and-middle-income countries, is now starting to recognise the extent and severity of the disease [3]. Breast cancer is the second most common cancer irrespective of gender in Bangladesh, and it is the leading cancer (29.3%) in females, according to a hospital-based cancer registry report [12]. Among the non-cancerous breast lesions, fibroadenomas are also more frequently reported in females of Bangladesh [13]. In this prevailing condition, there is still a paucity of information regarding the molecular characteristics of these breast disorders, including the expression of BRCA1 mRNA in Bangladeshi females.
The aim of this study was to measure the expression of BRCA1 mRNA in cancerous and non-cancerous breast tissue of Bangladeshi females. Thus, this research can provide the opportunity to improve the understanding of the molecular behaviour of breast cancer as well as non-cancerous breast lesions in this population for adopting appropriate therapeutic measures and predicting the prognosis of these diseases.
Bangladeshi females with breast cancer and benign breast disorders attending NICRH participated in this research. The names and mobile numbers of the histologically diagnosed breast cancer patients and non-cancerous females with benign breast lesions from January 2021 to September 2022 were collected from the register of the Department of Histopathology of NICRH. Inclusion criteria were Bangladeshi female, aged 18 years or above, (i) for breast cancer patient: histologically diagnosed as breast cancer patients; (ii) for females with benign breast disorders: histologically diagnosed as non-cancerous patients with benign breast lesions. Exclusion criteria were having a history of other cancers and chemo or radiotherapy before FFPE block preparation.
Formalin-fixed paraffin-embedded (FFPE) cancerous and non-cancerous breast tissue blocks were collected from the Department of Histopathology of NICRH. A list of 915 breast cancer patients and 126 females with non-cancerous breast lesions was obtained during this period. Among them, 62 breast cancer patients (based on recently prepared tissue blocks) and 27 non-cancerous females (out of 68 available FFPE tissue blocks which had sufficient breast tissue) with benign breast lesions were invited to participate in this study. Fifty (50) breast cancer patients and 19 non-cancerous females were finally selected from them based on quality and quantity of cancerous and non-cancerous breast tissue in the paraffin blocks (assessed by the histopathologist). Reproductive and cancer-related characteristics were collected from the participants through interviews and from hospital and medical records. BRCA1 mRNA expression in cancerous and non-cancerous breast tissue was measured and compared.

Types of breast tissue | BRCA1 expression status, number (%) | |
Expressed (n=16) | Not expressed (n=34) | |
Cancerous tissue (n=50) | 16 (32.0) | 34 (68.0) |
Non-cancerous tissue (n=19) | 7 (36.8) | 12 (63.2) |
Fibroadenoma | 3 (16.0) | 7 (37.0) |
Fibrocystic disease | 1 (5.0) | 1 (5.0) |
Ductal hyperplasia | - | 1 (5.0) |
Fibrocystic disease with ductal hyperplasia | 1 (5.0) | 3 (16.0) |
Normal breast tissue | 2 (10.5) | - |
Type of breast tissue | BRCA1 expression status, number (%) | |
Expressed (n=16) | Not expressed (n=34) | |
Cancerous tissue (n=50) | 16 (32.0) | 34 (68.0) |
Non-cancerous tissue (n=19) | 7 (36.8) | 12 (63.2) |
Fibroadenoma | 3 (16.0) | 7 (37.0) |
Fibrocystic disease | 1 (5.0) | 1 (5.0) |
Ductal hyperplasia | - | 1 (5.0) |
Fibrocystic disease with ductal hyperplasia | 1 (5.0) | 3 (16.0) |
Normal breast tissue | 2 (10.5) | - |

In this study, the majority of the breast cancers were diagnosed at or below the age of 45 years. Most of the cancers were sporadic or non-familial (86%), family history of cancers was reported in seven patients. Lymph node metastasis was present in almost all patients (96%), whereas distant metastasis was reported in 4 (8%) patients. Hormone receptor negativity was reported in most of the patients (Table 2).
Reproductive and cancer-related characteristic | BRCA1 expression status, number (%) | P | |
Expressed (n=16) | Not expressed (n=34) | ||
Age at diagnosis (years) | |||
28–45 | 8 (50.0) | 26 (76.5) | 0.06 |
46–69 | 8 (50.0) | 8 (23.5) |
|
Body mass index (kg/m2) | |||
18.5–24.9 | 11 (68.8) | 19 (55.9) | 0.39 |
≥25 | 5 (31.2) | 15 (44.1) |
|
Age at menarche (years) | |||
12–13 | 15 (93.8) | 26 (76.5) | 0.24 |
14–15 | 1 (6.2) | 8 (23.5) |
|
Age at menopause (years) (n=19) | |||
37–44 | 2 (25.0) | 5 (45.5) | 0.63 |
45–58 | 6 (75.0) | 6 (54.5) |
|
History of consanguinity of marriage | |||
Yes | 4 (25.0) | 1 (2.9) | 0.03 |
No | 12 (75.0) | 33 (97.1) |
|
Number of children |
|
|
|
1–2 | 11 (68.8) | 26 (76.5) | 0.73 |
3–5 | 5 (31.2) | 8 (23.5) |
|
Duration of breastfeeding (years) | |||
1–2 | 4 (25.0) | 7 (20.6) | 0.73 |
>2 | 12 (75.0) | 27 (79.4) |
|
Duration of contraceptive use (years) | |||
Never used | 1 (6.2) | 6 (17.6) | 0.21 |
1–5 | 7 (43.8) | 19 (55.9) |
|
≥6 | 8 (50.0) | 9 (26.5) |
|
Family history of cancer | |||
Positive | 1 (6.2) | 7 (20.6) | 0.41 |
Negative | 15 (93.8) | 27 (79.4) |
|
Lymph node metastasis | |||
Present | 15 (93.8) | 33 (97.1) | 0.54 |
Absent | 1 (6.2) | 1 (2.9) |
|
Other organ metastasis | |||
Present | 1 (6.2) | 3 (8.2) | 0.99 |
Absent | 15 (93.8) | 31 (91.8) |
|
Her-2/Neu (n=49) | |||
Positive | 5 (33.3) | 9 (26.5) | 0.62 |
Negative | 10 (66.7) | 25 (73.5) |
|
Oestrogen receptor (n=49) | |||
Positive | 4 (26.7) | 5 (14.7) | 0.43 |
Negative | 11 (73.3) | 29 (85.3) |
|
Progesterone receptor (n=49) | |||
Positive | 3 (20.0) | 5 (14.7) | 0.69 |
Negative | 12 (80.0) | 29 (85.3) |
|
Reproductive characteristic | Breast cancer patient (n=50) | Non-cancerous female (n=19) | P |
Age (years)a | 43.0 (9.5) | 33.5 (10.0) | 0.001 |
Body mass index (kg/m2)a | 24.4 (3.5) | 24.2 (3.0) | 0.98 |
Age at menarche (years)a | 12.8 (0.8) | 13.0 (0.7) | 0.80 |
Menstrual status, n (%) | |||
Menstruating | 31.0 (62.0) | 16.0 (84.2) | 0.13 |
Postmenopausal | 19.0 (38.0) | 3.0 (15.8) |
|
Age of menopause (years)a | 45.2 (4.6) | 43.3 (5.7) | 0.63 |
Consanguinity of marriage, n (%) | |||
Yes | 5.0 (10.0) | 1.0 (5.3) | 0.99 |
No | 45.0 (90.0) | 18.0 (94.7) |
|
Number of childrenb | 2.0 (1.0–5.0) | 2.0 (1.0–3.0) | 0.56 |
Total duration of breast feeding (years)b | 3.0 (1.5–8.0) | 3.0 (2.0–6.5) | 0.79 |
Use of contraceptives, n (%) | |||
No | 7.0 (14.0) | 4.0 (21.0) | 0.48 |
Yes | 43.0 (86.0) | 15.0 (79.0) |
|
Duration of contraceptive use (years)b | 5.0 (1.0–20.0) | 6.0 (1.0–14.0) | 0.28 |
aMean and standard deviation; bMedian and range |
BRCA1 protein is required for the prevention of breast transformation. BRCA1 expression status in non-cancerous benign breast lesions indicates the malignant potential of the breast. Non-expression or reduced expression of this gene in non-cancerous benign lesions requires close monitoring and follow-up, as inactivation of BRCA1 causes blunted ductal development, breast hyperplasia and tumour formation [19]. We have few non-cancerous patients other than fibroadenoma. They were histologically diagnosed as fibrocystic disease, ductal hyperplasia and the concomitant presence of fibrocystic disease with ductal hyperplasia. Non-expression of BRCA1 mRNA was observed more in patients with concomitant presence of fibrocystic disease with ductal hyperplasia in our study. This finding also warns us to keep close monitoring of the patients with non-cancerous benign breast disorders.
In our study, BRCA1 was expressed in 32% of breast cancer patients. Al Mullah et al. observed BRCA1 mRNA expression in 21% of EEPE tissue block. They also observed that the reduced or no expression of BRCA1 mRNA was more in the high histological grade of the tumour [20]. Taylor et al. found reduced expression of BRCA1 in non-familiar breast cancer and non-expression in 20% of non-familiar and 20% of familial breast cancer patients. They conducted the research on 142 non-familiar, 25 familial breast cancer patients and 28 non-cancerous females with fibrocystic disease, fibroadenoma and normal breast tissue [21]. In their study, mutation was reported in breast cancer patients with a family history of cancers and loss of heterozygosity (LOH) was reported in non-familial patients. If the normal functional allele of BRCA1gene is lost in LOH, this gene will exhibit non-expression. Silvia et al. reported 47% of LOH in the BRCA1 region (17q21) in patients with breast cancer [22]. In addition to LOH, hypermethylation is another cause of non-expression of the BRCA1 gene in breast cancer. The CpG islands of the promoter regions of tumour suppressor genes are usually unmethylated, and hypermethylation or abnormal methylation of these sites causes non-expression of those genes [23]. Hypermethylation at these sites of BRCA1 causes the inactivation of this gene, known as LOH. Esteller et al. found hypermethylation in the promoter region of the BRCA1 gene in 13% of primary breast cancer. That study also noticed abnormal methylation was more common in patients with breast cancer at or below the age of 45 years, and it was present in grade II or III invasive ductal carcinoma [24]. Therefore, considering the cancer-related characteristics of our participants, we can assume abnormal methylation or LOH might be a cause of reduced or non-expression of BRCA1 mRNA in our patients.
Taylor et al. did not find any correlation of BRCA1 expression with age, menopausal status, hormonal status, histological type and tumor size [21]. Li et al. also did not find any association between BRCA1 mRNA expression and clinicopathological characteristics of the breast cancer patients [5]. Our finding is consistent with the findings of these literature.
In our study association was found betweenBRCA1 mRNA expression and breast cancer patients with family history of consanguinity. The progeny of the consanguineous couples has more chance to get homozygous alleles. Thus, the frequency of genetically determined diseases in offspring of consanguineous parents is influenced by the homozygosity of the genes responsible for that conditions [25]. The frequency of breast cancer among the daughters of consanguineous parents varies in different populations. Studies on Pakistani, Croatian and North American population found that the younger women of parents with first-cousin marriages have increased risk of developing breast cancer, whereas, several studies conducted on Arab, North African and non-Jew Israeli population found low incidence of breast cancer in daughters of consanguineous parents [26–28]. Medimegh et al. conducted a study on Tunisian breast cancer patients and did not find homozygosity in BRCA1 haplotypes in consanguineous patients with familial breast cancer. They reported more BRCA1 haplotypes homozygotes in healthy consanguineous controls and more heterozygotes in non-consanguineous group. They explained their findings by the facts that there were three possible genotypes in consanguineous families: the first- without mutations gave healthy progeny, the second- with deleterious mutations at heterozygote state and caused breast cancer and the third- the deleterious mutations at homozygote state which was lethal [25]. Mice model studies proved the lethal effects of BRCA1 knockout homozygotes [29, 30]. Moreover, a computer simulated study on the consequences of long term practice of consanguineous marriage on the prevalence of lethal cancer genes found that mutation carrier rate of BRCA1/2 genes decreased six times faster in a highly consanguineous population than non-consanguineous population if spontaneous abortion and gene flow were considered to be absent [31]. From the above-mentioned findings we can assume that the patients from consanguineous parents have factors other than BRCA1 gene responsible for tumourogenesis.
Estimation of BRCA1 expression in breast cancer is important for selection of effective chemo and targeted therapy. Studies found that the cancer cells deficient of BRCA1 or BRCA2 genes are more sensitive to PARP1 inhibitors whereas the cells with normal BRCA expression are unaffected by these drugs [32]. Estimation of BRCA1 expression is required for BRCA replacement therapy.