Histopathologic Feature and Vitamin D Receptor Expression of

Benign Breast Tumor

Mega Sari Sitorus

, Dwi Rita Anggraini

, Lita Feriyawati

Departement of Anatomy, Medical Faculty, Universitas Sumatera Utara

Keywords: VDR, Benign Breast Tumor, Fibroadenoma.

Abstract: Many studies say that vitamin D levels are useful as a protection that can decrease the risk of breast cancer,

colon, prostate and ovarian cancers. But some say it's not related. But how the expression of benign breast

tumor never been reported. The existence of VDR can be showed by its expression through

immunohistochemistry. This report will describe the expression of VDR in benign breasst tumor which the

most is fibroadenoma mamma. Pathological tissue samples were obtained from 50 patients suffered with

benign breast lesion that treated in Grandmed Hospital. There are 50 females, the mean age was 24,1, ranging

from 12 to 42 years of age. All of the Hematoksilin Eosin staining slides are examined by pathologist using

light microscope and concluded as fibroadenomas. The presence and location VDR is detected by

immunostaining with VDR Antibody (D-6) from Santa Cruz Biotechnology. One of the most common benign

tumors in the breast around 30 years old is fibroadenomas. In this study it is found that 34 patients (68%)

under 30 years old. The expression of VDR in the fibroadenomas are strong : 42 tumours (82%), moderate:

4(8%, low: 3 (6%), negative: 1(2%). Most of them are VDR positif expression. From the figure 4 we can see

that the VDR is placed in the nuclear dan cell membrane of the cells. Stromal cells doesn’t show expression

of VDR. This discribe that only epithelial cell of breast glandular this receptor.

1 INTRODUCTION

Usually, the tumor that grows in the breast are benign.

Attention is more given to malignant breast lesion

because breast cancer is the most common

malignancy among women in the world. Actually,

benign beast tumor are more frequent than malignant

lesions (Guray, 2006). Fibroadenoma of the breast is

a relatively frequently occurring tumor. It can occur

in any age of women, but the peak incidence is in

twenty until thirty tears of age. Although often

considered a benign tumor, several reports describe a

higher risk of subsequent breast carcinoma in patients

diagnosed with fibroadenoma (Kuijper, 2001).

The study during the two decades before has

mentioned that the variety biological actions of 1,25-

dihydroyxyvitamin D

(1,25(OH)

/ Calcitriol) are

triggered by exactly changes in gene expression

which are assisted by intracellular vitamin D receptor

(VDR)(Pike JW, 2010). Mostly of breast tumors

(58%) displayed moderate to strong VDR expression.

An aggressive tumor display inversely related to

VDR expression (A-Azhri, 2017).

A fat soluble vitamin D consist of steroid

molecular structures (Yosephine B, 2016). Sunlight

exposure is the main source of vitamin D for human.

Ultraviolet B light from the sun will penetrate the

skin, change 7-dehydrocholesterol from

keratinocytes and fibroblasts in plasma membranes to

be previtamin D3. In the plasma membrane,

previtamin D3 will be converted rapidly to vitamin

D3, then released into the extracellular space. In the

skin Vitamin D3 is bounded vitamin D-binding

protein. While vitamin D2 and vitamin D3 from the

diet therapy bound with vitamin D binding protein

and lipoprotein. In the liver, vitamin D2 and D3 are

hydroxylated to be 25-hydroxyvitamin D [25 (OH)

D]. 25 (OH) D is the main form of circulation. This

form molecule is used to measure the vitamin D

status. 25 (OH) D is still inactive form and should be

hydroxylized at carbon 1 to be 1,25-

dihydroxyvitamin D [1,25 (OH) 2D/calcitriol] by the

kidney and other tissues, including the breast (Katie

MO, 2017).

1,25 (OH) 2D has endocrine role to control

calcium metabolism by increasing absorption and

mobilizing calcium from bone (Holick MF, 2018).

880

Sitorus, M., Anggraini, D. and Feriyawati, L.

Histopathologic Feature and Vitamin D Receptor Expression of Benign Breast Tumor.

DOI: 10.5220/0010098208800883

In Proceedings of the International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches (ICOSTEERR 2018) - Research in Industry 4.0, pages

880-883

ISBN: 978-989-758-449-7

Other sources of vitamin D are very few gets from

foods including oily fish, such as salmon, mackerel,

tuna, and sardines. Vegetable foods generally contain

low vitamin D. Now there are many kinds of foods

which contain fortified vitamin D, particularly milk

products, biscuits, orange juice, yogurt and cereals.

Some foods, such as milk, bread, cereals, are fortified

with vitamin D (Yosephine B, 2016; Holick MF,

2018).

Actions of 1,25 (OH)2D are mediated by the VDR.

It consists of three domains: the C-terminal ligand

binding domain, N-terminal DNA binding domain,

and hinge region binding binds these two domains

together (Bikle DD, 2014). VDR belongs to the

nuclear subfamily receptors acts as transcription

factors into the target cells after making a dimer with

retinoid X receptors (RXR). After dimerization, the

compound molecule binds to VDR(Gil A, 2018). In

the intestine and bone this action to augment intestinal

calcium absorption and influence osteoclast activity.

Prostate, colon, and breast express 1-hydroxylase,

non-calcium regulating tissues locally convert

25(OH)D to 1,25(OH)2D which regulate cell

proliferation and differentiation, to possibly decrease

the risk of cell transformation into a malignant lesion.

Almost tissues and cells in the body have vitamin D

receptor. The presence of 1,25(OH)2D to be one of the

most potent regulators of cellular growth in normal

condition and cancer cells (Stechschulte SA, 2009).

Many studies say that vitamin D levels are useful

as a protection that can decrease the risk of colon

cancer, breast, prostate and ovarian cancers. But some

say it's not related (Gil A, 2018). But how the

expression of benign breast tumor never been

reported.

The existence of VDR can be showed by its

expression through immunohistochemistry. This

report will describe the expression of VDR in benign

breasst tumor which the most is fibroadenoma

mamma.

2 MATERIAL AND METHODS

Pathological tissue samples were obtained from 50

patients suffered with benign breast lesion that treated

in Grandmed Hospital. There are 50 females, the

mean age was 24,1, ranging from 12 to 42 years of

age. All of the Hematoksilin Eosin staining slides are

examined by pathologist using light microscope and

concluded as fibroadenomas. The presence and

location VDR is detected by immunostaining with

VDR Antibody (D-6) from Santa Cruz

Biotechnology.

Table 1: Characteristics of the patients.

Age of patients with fibroadenoma mammae

–

ears 2 (4.0%)

–

ears 10 (20.0%)

–

ears 18 (36.0%)

–

ears 4 (8.0%)

–

ears 7(14.0%)

–

ears 5(10.0%)

–

ears 4(8.0%)

Immunohistochemistry and Evaluation

In this study, the samples are from formalin-fixed

paraffin embedded pathological tissue. The VDR

expression were studied by immunostaining using

monoclonal antibodies VDR Antibody (D-6) from

Santa Cruz Biotechnology, according the standards

protocols used in laboratories of Murni Teguh

Hospital .

VDR immunostaining intensity of fibroadenoma

was examined semiquantitatively by two independent

observers from 0 to 3 arbitrary units with 0 as

negative (0), weak/low (1), moderate (2) and strong

(Pike JW, 2010). VDR staining intensity in

fibroadenoma lesions was evaluated with reference to

intense reddish-pink basal layer of normal skin

epidermis, scored as strong. Light reddish-pink and

light pink stained cells were scored as cells with

moderate or weak VDR expression, correspondingly.

The VDR expression was assessed for cytoplasm and

nuclei of cells separately (Brozyna AA, 2011).

Figure 1: Representative images of vitamin D receptor

(VDR) protein expression, classified by immunoreactive

score (IRS) (Al-Zahri J, 2017).

3 RESULTS

The most common benign tumors of the breast in

woman is fibroadenomas. This tumor usually attack

woman under 30 years old. In this study it is found

that 34 patients (68%) under 30 years old.

Microscopically, fibroadenoma is a benign tumor

with biphasic features consist of epithelial and

stromal components (Lee M, 2015).

The microscopic features showed in figure 2. The

tumour consists of epithelial component as a

Histopathologic Feature and Vitamin D Receptor Expression of Benign Breast Tumor

881

glandular slit or tubular form and propiferative

stromal component.

Figure 2: Pathologic view of fibroadenomas.

Most of the of fibroadenomas present as a single

mass (70%). Others are as multiple masses around

10%–25%. Characteristic of fibroadenomas are a

painless, mobile, smooth, and rubbery mass with

distinct borders. The size usually ranging from 1 cm

to 3 cm, on the upper outer quadrant of the breast. It

can also be very small unpalpable mass that it is only

seen on microscopic examination. Simetimes can be

larger than 10 cm and may cause breast asymmetry

and significant esthetic problem of the breast. The

finding existence of VDR in other tissue beside gut

and skeletal are observed in this study in benign

breast tumor (figure 3).

Figure 3. VDR expressions in fibroadenomas, A. Negative,

B. Low, C. Moderate, E. Strong (100x).

4 DISCUSSION

Fibroadenomas is the most benign breast tumour in

woman. Usually they are found in young middle age,

below 30 years old. In this study found that 68%

tumours are found under 30 years old. As a benign

breast tumor, fibroadenoma is also thought to

represent a set of hyperplastic breast lobules called

“aberrations of normal development and involution”.

This lesion is a hormone-dependent neoplasm that

lactates during pregnancy and also get involuting

along with the rest of the breast in perimenopause.

Figure 4. Distribution of VDR Expression (100x).

The expression of VDR in the fibroadenomas are

strong : 42 tumours (82%), moderate: 4(8%, low: 3

(6%), negative: 1(2%). Most of them are VDR positif

expression. From the figure 4 we can see that the

VDR is placed in the nuclear dan cell membrane of

the cells. Stromal cells doesn’t show expression of

VDR. This discribe that only epithelial cell of breast

glandular this receptor.

5 CONCLUSION

The histopathologic feature of benign breast tumor

mostly is fibroadenoma mammae. The glandular dan

stromal cells proliferates resemble the cleft like

appearence. With imunohistochemistry can detect

that vitamin D receptor also expressed in the

epithelial cells, both in nuclei and cytoplasm.

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