IMMUNOHISTOCHEMICAL AND HISTOPATHOLOGICAL ASSAYS OF WATER-SOLUBLE PEPTIDES (WSPS) BASED CYTOTOXINS IN DMBA-INDUCED BREAST CANCER RAT MODELS

Authors: Sana Mahmood,  Mujahid ul Islam, Tayyaba Tariq, Farwa Tariq,, Aqsa Parveen, Muhammad Issa Khan, Muhammad Naeem Faisal  and  Aysha Sameen,
Journal: Journal of Animal and Plant Sciences (JAPS)
ISSN: 1018-7081 (Print), 2309-8694 (Online)
Volume: 36  Issue: 4  
Year: 2026
DOI: https://doi.org/10.36899/JAPS.2026.4.0095
URL: https://doi.org/https://doi.org/10.36899/JAPS.2026.4.0095
Publisher: Pakistan Agricultural Scientists Forum

Abstract:
<p style="text-align: justify;"><span style="font-size: 12.0pt; mso-bidi-font-size: 14.0pt; line-height: 115%; font-family: 'Times New Roman',serif; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">Bioactive peptides-based drugs, being a novel therapeutic approach as an alternative to conventional chemotherapy, are the next-generation option. In this milieu, the present study was designed to evaluate the anti-tumoral activities of Water-Soluble Peptides (WSPs) derived from casein, whey, fish bones, and mixed WSPs in 7,12-dimethylbenz (a) anthracene (DMBA)</span><span style="font-size: 12.0pt; mso-bidi-font-size: 10.0pt; line-height: 115%; font-family: 'Times New Roman',serif; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">-</span><span style="font-size: 12.0pt; mso-bidi-font-size: 14.0pt; line-height: 115%; font-family: 'Times New Roman',serif; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">induced mammary carcinoma. The mammary carcinoma was induced in rats by administering 60mg/kg DMBA by dividing into two doses intra-gastrically through gavage. The rats were divided into 6 groups: G<sub>0</sub> (Negative control), G<sub>1</sub> (Doxorubicin receiving rats), G<sub>2</sub> (<span style="mso-bidi-font-weight: bold;">Casein WSPs extract)</span>, G<sub>3</sub> (<span style="mso-bidi-font-weight: bold;">Whey WSPs extract)</span>, G<sub>4</sub> (<span style="mso-bidi-font-weight: bold;">Fish bones WSPs extract),</span> and G<sub>5</sub> (<span style="mso-bidi-font-weight: bold;">Mixed WSPs). </span>Physical parameters and biochemical assays were performed at three intervals (<span style="color: black; mso-color-alt: windowtext; background: white;">1<sup>st</sup>, 6<sup>th,</sup> and 12<sup>th</sup> week)</span>. The WSPs significantly (<em>p</em></span><span style="font-size: 12.0pt; mso-bidi-font-size: 14.0pt; line-height: 115%; font-family: 'Times New Roman',serif; mso-fareast-font-family: STIX-Regular; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">&lt;</span><span style="font-size: 12.0pt; mso-bidi-font-size: 14.0pt; line-height: 115%; font-family: 'Times New Roman',serif; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">.001) improved the activity of antioxidant enzymes in carcinogenic animals. Furthermore, the fish and mixed WSPs also prevented the increase in the concentration of tumor biomarkers that are carcinoembryonic antigen (CEA), erythrocyte sedimentation rate (ESR), and p53 in experimental rats. Continual increase in liver function enzymes was also observed in G<sub>1</sub> and G<sub>2 </sub>as compared to G<sub>4</sub> and G<sub>5</sub>. The <span style="color: black; mso-color-alt: windowtext; background: white;">immunohistochemical analysis was also conducted to assess the expression of proteins related to epigenetic alterations. The results revealed that p53 protein expression decreased in </span>G<sub>1</sub> and G<sub>5,</sub> whereas the expression of breast cancer gene-1 (BRCA1) and breast cancer gene-2 (BRCA2) increased in G<sub>1</sub>, G<sub>4,</sub> and G<sub>5</sub> due to increased immunoreactivity and well-defined nuclear staining. This can be concluded from the findings that G<sub>4</sub> and G<sub>5</sub> exhibited the strong hepatic toxicity mitigation potential as compared to G<sub>1</sub>. This study opens the window to consider bioactive peptides for the </span><span style="font-size: 12.0pt; mso-bidi-font-size: 14.0pt; line-height: 115%; font-family: 'Times New Roman',serif; mso-fareast-font-family: Georgia; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">development of functional foods or new drugs for anticancer therapy.</span></p>

Keywords: neoplasm, memory tissues, oxidative stress, natural therapies, bioactive peptides, therapeutic potential