Elsevier

Life Sciences

Volume 195, 15 February 2018, Pages 71-80
Life Sciences

Tumor control by hypoxia-specific chemotargeting of iron-oxide nanoparticle – Berberine complexes in a mouse model

https://doi.org/10.1016/j.lfs.2017.12.036Get rights and content

Abstract

Aim

To evaluate the therapeutic efficacy of hypoxic cell-sensitizer Sanazole (SAN) –directed targeting of cytotoxic drug Berberine (BBN) and Iron-oxide nanoparticle (NP) complexes, to solid tumor in Swiss albino mice.

Main methods

NP-BBN-SAN complexes were characterized by FTIR, XRD, TEM and Nano-size analyzer. This complex was orally administered to mice-bearing solid tumor in hind limb. Tumor regression was analysed by measuring tumor volume. Cellular DNA damages were assessed by comet assay. Transcriptional expression of genes related to tumor hypoxia and apoptosis was evaluated by quantitative real-time PCR and morphological changes in tissues were analysed by histopathology. Also levels of antioxidants and tumor markers in tissues and serum biochemical parameters were analysed.

Key findings

Administration of NP-BBN-SAN complexes reduced tumor volume and studies were focussed on the underlying mechanisms. Extensive damage to cellular-DNA; down-regulated transcription of hif-1α, vegf, akt and bcl2; and up-regulated expression of bax and caspases, were observed in tumor. Results on tumor markers, antioxidant-status and serum parameters corroborated the molecular findings. Histopathology of tumor, liver and kidney revealed the therapeutic specificity of NP-BBN-SAN.

Significance

Thus SAN and NP can be used for specific targeting of drugs, to hypoxic solid tumor, to improve therapeutic efficacy.

Graphical abstract

The administration of NP-BBN-SAN complexes is found to down regulate the expression of genes responsible for hypoxia-induced tumor progression. Further the complexes initiates the up regulation of apoptotic genes' expression. It causes apoptosis in tumor, which is further evidenced from histopathology, and leads to tumor regression. SAN has the capability to accumulate in hypoxic area of tumor, hence it directs the cytotoxic drug BBN in the complexes to tumor to facilitate tumor regression. Histopathology of Liver and Kidney of these animals are appeared normal which is a major evidence for treatment specificity.

NP- Iron-oxide nanoparticles, BBN- Berberine, SAN- Sanazole, hif-1alpha: hypoxia inducible factor-1 alpha, vegf: vascular endothelial growth factor, akt. protein kinase B, bcl2: B-cell lymphoma 2, bax: bcl2-associated X protein, GPx: glutathione peroxidase, GSH: glutathione, SOD- superoxidde dismutase, MDA- malodialdehyde.

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Section snippets

Background

Success of chemotherapy is often limited by systemic toxicities and side effects. One of the means of overcoming these problems of antineopalstic agents is to deliver the drugs at the tumor site. Hypoxic region in tumor supports not only tumor growth but also resistance to major therapies such as chemotherapy and radiation. The major cellular adaptive response under the reduced oxygen state in tumor tissues is the expression of hypoxia-inducible factor-1 (HIF-1). HIF-1 is a DNA-binding protein

Chemicals

FeCl2, FeCl3, Chloroform and Isoamyl alcohol were bought from Merk, India. Berberine, chemicals for RNA isolation, β-D Glucuronidase, Brij and O-Dianisidine were purchased from Sigma Aldrich, India. Sanazole (AK2123) was obtained from Dr. V.T.Kagiya, Health Research Foundation, Kyoto, Japan. All other chemicals were obtained from reputed national manufactures (Otto Chemie Pvt. Ltd. and Himedia Pvt. Laboratories Ltd).

Animals

Female Swiss albino mice weighing 24-28 g were purchased from Small Animal

Characterization of NPs and NP-drug complexes

The characteristics of Fe3O4 nanoparticles (magnetite) prepared by co-precipitation method and its complexes with BBN and SAN were evaluated by FTIR, XRD, TEM and Size analysis. As shown in Fig. 1, FTIR measurements was recorded in the range of 500–4000 cm 1. The strong peak at 3404.59 cm 1 in NP was indicated the presence of –OH stretch in the nanoparticles. A peak observed around 2843 cm 1in BBN could be allotted to Csingle bondH stretching vibrations of methoxy group in BBN. The peak located around 1504 cm

Discussion

The uncontrolled rapid proliferation of tumor cells confers special microenvironment in the tissues of tumors due to inadequate vasculature, hypoxia, lower nutrient status as well as drainage of cellular metabolic toxins. This would manifest in altered gene expression and metabolism in tumor tissues, drug resistance and therapeutic availability of administered chemotherapeutics leading to dose escalation and the resultant systemic toxicity. Tumor targeted delivery of therapeutics is the best

Conclusion

NP-BBN-SAN complexes were found to be more effective in reducing tumor volume compared to the other treatments. The mechanism behind the tumor regression can be ascribed to i) the down regulation in the transcription of hif-1α and its associated genes- vegf and akt, thereby resulting in tumor regression and ii) tnf-α induced extrinsic pathway of apoptosis through caspase 8, although other apoptotic pathways also may be involved. The present study convincingly demonstrates clinical relevance and

Acknowledgements

The authors thank University Grants Commission (UGC), Government of India for senior research fellowship to SS. The authors thank Dr. M.O. Annamma, Head, Department of Pathology, Pushpagiri Institute of Medical Sciences and Research Centre, Thiruvalla for histopathology analysis, Sreekanth K Mahadeva and K V Rao, Department of Materials Science and Engineering, Royal Institute of Technology Stockholm, Sweden; Dr. Jacob Varkey, Rubber Research Institute, Kottayam and Dr. Nandakumar, School of

Funding source

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors except senior research fellowship from University Grants Commission, Government of India to one of the authors (SS).

Conflict of interest

None.

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