Heparinase I/III mixture induces cytotoxicity in cancer cell lines via oxidative stress, DNA damage, and activation of tumor suppressor genes: Anticancer activity of heparinase

Nawaf Khalid Mohammed; Bahaa Abdullah Laftaah Al-Rubaai

doi:10.62310/liab.v6i2.383

Authors

Nawaf Khalid Mohammed Department of Biology, College of Science, University of Baghdad, Baghdad, Iraq
Bahaa Abdullah Laftaah Al-Rubaai Department of Biology, College of Science, University of Baghdad, Baghdad, Iraq

DOI:

https://doi.org/10.62310/liab.v6i2.383

Keywords:

Heparinase, Cytotoxicity, Heparin sulfate, A549 cells, DNA damage, Tumor supressor genes

Abstract

Heparan sulfate, an integral component of the extracellular matrix, plays a vital role in the tumor progression. Hence targeting the heparan sulfate-mediated signaling pathway offers a promising strategy in anticancer therapy. The present study was conducted to assess the anticancer activity of 1:1 mixture of heparinase I and heparinase III against three cancer cell lines, including MCF-7 breast cancer, A549 lung cancer, and AsPC-1 pancreatic cancer with respect to the normal fibroblast cell line. The study explored the effects of heparinase on the cytotoxicity, oxidative stress, DNA damage, and expression of tumor suppressor genes in the cell lines. The results demonstrated significant cytotoxicity, dependent on the cell type, with A549 cells exhibiting the highest sensitivity (IC50 = 0.049 U/mL) compared to normal fibroblast cells (IC50 = 0.13 U/mL). Moderate cytotoxicity was observed in AsPC-1 cells, whereas limited cytotoxicity was displayed by MCF-7 cells. Significant morphological alterations in AsPC-1 cells, characterized by cell rounding, shrinkage, and reduced density, were observed in response to heparinase treatment compared to untreated control cells. A significant increase in ROS levels was observed in treated cells, suggesting oxidative stress in exposed cells. Furthermore, comet assay and qPCR revealed significant DNA damage and upregulation of tumor suppressor genes – p53 (8.46 fold) and p16 (4.23 fold), respectively, in response to heparinase treatment of A549 cells. In conclusion, the results demonstrate the anticancer activity of heparinase mixture through oxidative stress, DNA damage, upregulation of tumor suppressor genes and associated pathways. Though this study provides valuable insights into the anticancer therapeutic potential of heparinase, further in vivo studies are required with validation at the protein level to establish the therapeutic relevance of anticancer properties of this enzyme.

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