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Tannic acid enhances cisplatin effect on cell proliferation and apoptosis of human osteosarcoma cell line (U2OS)

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Abstract

Background

The increase in cases of chemoresistance of cisplatin for osteosarcoma treatment has called for the need to establish a new treatment regime. Tannic acid (TA) possesses a potent antiproliferative effect against various cancers. Therefore, this study investigated the effect of TA combined with cisplatin on human osteosarcoma cell lines (U2OS).

Methods

MTT assay was used to determine the half-maximal inhibitory concentration (IC50), while the combination index (CI) value was utilized to analyze the interaction within each combination. The antiproliferative effect of the treatment was evaluated by trypan blue exclusion assay. The morphological changes of cells were observed under a phase-contrast inverted microscope. The nuclear morphology and percentage of apoptosis cells were evaluated by using the Hoechst 33258 staining and annexin V/PI assay, respectively.

Results

The U2OS cells showed cytotoxic effect when treated with TA and cisplatin, with IC50 at 4.47 µg/mL and 16.25 µg/mL, respectively. The TA demonstrated no significant inhibition effect on the normal human fetal osteoblast cells (hFOB 1.19); yet, interestingly, a potent proliferative effect was indicated. Synergistic interaction was triggered when TA was combined with cisplatin at percentage ratios of 90:10 and 85:15. Meanwhile, antagonistic interaction was induced in the combination at percentage ratios of 75:25 and 50:50. On the other hand, a significant antiproliferative effect with prominent morphological alteration was detected in the cells treated with a combination of TA and cisplatin at the percentage ratio of 90:10. Additionally, combination-treated cells demonstrated the highest percentage of apoptosis cells, with distinct chromosomal condensation, nuclear fragmentation, reduction of nuclear volume, and notable apoptotic body.

Conclusion

Therefore, there is a high potential for the inclusion of TA in the cisplatin-based chemotherapeutic regimen of osteosarcoma.

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Abbreviations

CI:

Combination index

CO2 :

Carbon dioxide

Cells/mL:

Cells per milliliter

FITC:

Fluorescein isothiocyanate

FACS:

Fluorescence-activated cell sorting

hFOB 1.19:

Human fetal osteoblast cells

IC50 :

Half-maximal inhibitory concentration

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

mg/mL:

Milligram per milliliter

µg/mL:

Microgram per milliliter

OD:

Optical density

PI assay:

Propidium iodide assay

ROS:

Reactive oxygen species

Saos-2:

Sarcoma osteogenic-2

TA:

Tannic acid

U2OS:

Human osteosarcoma cells

UV:

Ultraviolet

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Acknowledgements

The authors would like to express: special thanks to the Universiti Sains Malaysia (USM) for providing the fund under the Research University Grant (RUI) (1001/PPSK/8012318); sincere appreciation also to the Craniofacial Laboratory, School of Dental Sciences, and Biomedicine Laboratory, School of Health Sciences USM for providing the research facility to complete this study; special thanks also to Associate Professor Dr Wan Muhamad Amir Wan Ahmad from School of Dental Sciences USM for providing assistance in conducting statistical analysis of this study.

Funding

This study was supported by Research University Grant (RUI) (1001/PPSK/8012318) Universiti Sains Malaysia.

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Authors and Affiliations

  1. School of Health Sciences, Universiti Sains Malaysia, Health Campus, 16150 Kubang Kerian, Kelantan, Malaysia

    Mohamad Zahid Kasiram, Hermizi Hapidin, Hasmah Abdullah & Nor Munira Hashim

  2. School of Dental Sciences, Universiti Sains Malaysia, Health Campus, 16150 Kubang Kerian, Kelantan, Malaysia

    Ahmad Azlina

  3. School of Medical Sciences, Universiti Sains Malaysia, Health Campus, 16150 Kubang Kerian, Kelantan, Malaysia

    Sarina Sulong

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  1. Mohamad Zahid Kasiram
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Contributions

Study conception or design: HH and HA; data processing, collection, perform experiment: MZK and NMH; analysis and interpretation of results: MZK and NMH; draft manuscript preparation, visualization: MZK; critical revision of the paper: HH, HA, AA and SS; supervision, funding acquisition: HH; final approval of the version to be published: MZK, HH, HA, NMH, AA, and SS.

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Correspondence to Hermizi Hapidin.

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Kasiram, M.Z., Hapidin, H., Abdullah, H. et al. Tannic acid enhances cisplatin effect on cell proliferation and apoptosis of human osteosarcoma cell line (U2OS). Pharmacol. Rep 74, 175–188 (2022). https://doi.org/10.1007/s43440-021-00330-3

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