Natural bioactive 4-Hydroxyisophthalic acid (4-HIPA) exhibited antiproliferative potential by upregulating apoptotic markers in in vitro and in vivo cancer models


There is tremendous scope for identifying novel anti-cancer molecules from the unexplored reserves of plant kingdom. The application of dietary supplementation or medicine derived from such sources is a promising approach towards treatment of cancer. In the present study we have evaluated the antiproliferative potential of 4-hydroxyisophthalic acid (4-HIPA), which is a novel antioxidant compound isolated from the roots of the aqueous extract of Decalepis hamiltonii. 4-HIPA was screened in vitro against human breast cancer cell lines MCF-7, MDA-MB-468 and normal human breast epithelial cell MCF-10, and demonstrated that human breast cancer cell lines, in contrast to MCF-10, are sensitive to 4-HIPA .4-HIPA showed marked reduction in cell viability and short-term proliferation assays in these cells. Results of the long-term colony formation and scratch assay further reaffirmed that 4-HIPA inhibited the growth and proliferation in breast cancer cells. We further conducted in vivo studies using murine Ehrlich Ascites Tumor (EAT) cell model. Our in vivo results established that treatment with 4-HIPA reduced the tumorigenesis by promoting apoptosis in EAT-bearing mice. The results of our molecular docking predictions further warranted our claim. This study is valuable as 4-HIPA exhibits antiproliferative potential that can be exploited in the development of anticancer drugs.

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4-Hydroxyisophthalic acid




Ehrlich ascites tumor cells


Lactate dehydrogenase


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  1. 1.

    Wong MC et al (2016) Global Incidence and mortality for prostate cancer: analysis of temporal patterns and trends in 36 countries. Eur Urol 70(5):862–874

    Article  Google Scholar 

  2. 2.

    Zarei M, Shivanandappa T (2016) Neuroprotective effect of Decalepis hamiltonii on cyclophosphamide-induced oxidative stress in the mouse brain. J Basic Clin Physiol Pharmacol 27(4):341–348

    CAS  PubMed  Google Scholar 

  3. 3.

    Harish R et al (2005) Isolation of antioxidant compounds from the methanolic extract of the roots of Decalepis hamiltonii (Wight and Arn). J Agric Food Chem 53(20):7709–7714

    CAS  Article  Google Scholar 

  4. 4.

    Zarei M, Shivanandappa T (2013) Amelioration of cyclophosphamide-induced hepatotoxicity by the root extract of Decalepis hamiltonii in mice. Food Chem Toxicol 57:179–184

    CAS  Article  Google Scholar 

  5. 5.

    Srivastava A, Shivanandappa T (2006) Hepatoprotective effect of the aqueous extract of the roots of Decalepis hamiltonii against ethanol-induced oxidative stress in rats. Hepatol Res 35(4):267–275

    Article  Google Scholar 

  6. 6.

    Srivastava A, Rao LJM, Shivanandappa T (2012) A novel cytoprotective antioxidant: 4-Hydroxyisophthalic acid. Food Chem 132(4):1959–1965

    CAS  Article  Google Scholar 

  7. 7.

    Subhashree AR et al (2012) The reference intervals for the haematological parameters in healthy adult population of chennai, southern India. J Clin Diagn Res 6(10):1675–1680

    Google Scholar 

  8. 8.

    Kornberg A, Lieberman I, Simms ES (1955) Enzymatic synthesis of purine nucleotides. J Biol Chem 215(1):417–427

    CAS  PubMed  Google Scholar 

  9. 9.

    Lala V, Minter DA. (2019). Liver function tests, in StatPearls, StatPearls Publishing StatPearls Publishing LLC.: Treasure Island (FL)

  10. 10.

    Lala V, Minter DA. (2019). Acute cystitis, in StatPearls, StatPearls Publishing StatPearls Publishing LLC.: Treasure Island (FL)

  11. 11.

    Ali SJ et al (2019) Bone loss in MPTP mouse model of Parkinson's disease is triggered by decreased osteoblastogenesis and increased osteoclastogenesis. Neurotox Res 363:154–163

    CAS  Google Scholar 

  12. 12.

    Ali SJ, et al. (2019). Chlorpyrifos exposure induces parkinsonian symptoms and associated bone loss in adult swiss albino mice

  13. 13.

    Payne J et al (2000) Improving the reproducibility of the MCF-7 cell proliferation assay for the detection of xenoestrogens. Sci Total Environ 248(1):51–62

    CAS  Article  Google Scholar 

  14. 14.

    Franken NA et al (2006) Clonogenic assay of cells in vitro. Nat Protoc 1(5):2315–2319

    CAS  Article  Google Scholar 

  15. 15.

    Liang CC, Park AY, Guan JL (2007) In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2(2):329–333

    CAS  Article  Google Scholar 

  16. 16.

    Rates SM (2001) Plants as source of drugs. Toxicon 39(5):603–613

    CAS  Article  Google Scholar 

  17. 17.

    Ramnath V, Kuttan G, Kuttan R (2002) Antitumour effect of abrin on transplanted tumours in mice. Indian J Physiol Pharmacol 46(1):69–77

    CAS  PubMed  Google Scholar 

  18. 18.

    Zarei M et al (2013) Cardioprotective effect of the root extract of Hemidesmus indicus against doxorubicin-induced oxidative stress in mice. Pharm Lett 5(1):334–339

    Google Scholar 

  19. 19.

    Atanasov AG et al (2015) Discovery and resupply of pharmacologically active plant-derived natural products: a review. Biotechnol Adv 33(8):1582–1614

    CAS  Article  Google Scholar 

  20. 20.

    Harish R, Shivanandappa T (2010) Hepatoprotective potential of Decalepis hamiltonii (Wight and Arn) against carbon tetrachloride-induced hepatic damage in rats. J Pharm Bioallied Sci 2(4):341–345

    CAS  Article  Google Scholar 

  21. 21.

    Kogo M et al (2011) Analysis of the risk factors for myelosuppression after chemoradiotherapy involving 5-fluorouracil and platinum for patients with esophageal cancer. Hepatogastroenterology 58(107–108):802–808

    CAS  PubMed  Google Scholar 

  22. 22.

    Contreras-Zentella ML, Hernandez-Munoz R (2016) Is Liver enzyme release really associated with cell necrosis induced by oxidant stress? Oxid Med Cell Longev 2016:3529149

    Article  Google Scholar 

  23. 23.

    Srivastava JK, Gupta S (2006) Tocotrienol-rich fraction of palm oil induces cell cycle arrest and apoptosis selectively in human prostate cancer cells. Biochem Biophys Res Commun 346(2):447–453

    CAS  Article  Google Scholar 

  24. 24.

    Stefanini M (1985) Enzymes, isozymes, and enzyme variants in the diagnosis of cancer. A short review. Cancer 55(9):1931–1936

    CAS  Article  Google Scholar 

  25. 25.

    Xia M et al (2004) Dracorhodin perchlorate induces apoptosis via activation of caspases and generation of reactive oxygen species. J Pharmacol Sci 95(2):273–283

    CAS  Article  Google Scholar 

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The correspondent author wishes to thank Prof. Cletus D'Souza, the Head of the Department of Biochemistry, University of Mysore, for his support and guidance at the time of this study.

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Zarei, M., Shivanandappa, T. & Zarei, M. Natural bioactive 4-Hydroxyisophthalic acid (4-HIPA) exhibited antiproliferative potential by upregulating apoptotic markers in in vitro and in vivo cancer models. Mol Biol Rep (2020).

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  • Bioactive compound
  • 4-hydroxyisophthalic acid
  • Anti-tumor
  • pro-apoptosis
  • EAT model
  • Molecular docking