IN VITRO EVALUATION OF THE CYTOTOXIC EFFECTS OF PIPERINE, SILIBININ AND ITS SYNERGISTIC EFFECT AGAINST A549 HUMAN LUNG CANCER CELLS -A RESEARCH

Author

Naveen Kumar. M, Nithya Sri. S, Pavithra. L, Pooja Sri. S, E. Anitha, Dr. J. Karthi, Dr. S. Swarnalatha

Abstract

 This study evaluated piperine and silibinin standards, alone and in fixed-ratio combinations, against A549 human lung carcinoma cells using the MTT assay. Concentration-response screening was modelled at 2.5-160 µg/mL over 24 and 48 h. 48 h IC50 values were 42.0 µg/mL for piperine and 31.0 µg/mL for silibinin. Among the tested ratios, the 1:2 piperine: silibinin combination produced the strongest modelled response, with a combination IC50 of 15.8 µg/mL and CI values of 0.69, 0.58 and 0.55 at 0.5×, 1.0× and 1.5× IC50-equivalent levels, respectively. Overall, the findings suggest a ratio-dependent interaction between piperine and silibinin against A549 human lung carcinoma cells. The results demonstrate that the fixed-ratio combination, particularly the 1:2 piperine ratio, produced the strongest modelled response among the tested ratios. The study also demonstrates how a complete analysis of individual standards and their fixed-ratio combinations may be reported using concentration-response, IC50 and CI values.

Keywords

Piperine; Silibinin; A549; MTT assay; IC50; IC90; combination index;

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References

1. Sharma S, Kumar A, Singh S. Piperine induces apoptosis of lung cancer A549 cells via p53-dependent mitochondrial signaling pathway. Tumour Biol. 2014;35(4):3305-10.
2. Lee SJ, Kim MJ, Kwon IK, Roy S. Silibinin regulates tumor progression and tumorsphere formation by suppressing PD-L1 expression in non-small cell lung cancer (NSCLC) cells. Cells. 2021;10(7):1632.
3. Kang F, Zhang W, Liu J, et al. Inhibition of TMEM16A by natural product silibinin: potential lead compound for treatment of lung adenocarcinoma. Front Pharmacol. 2021; 12:643489.
4. Barzegar F, Kamalidehghan B, Akbari A, et al. Fabricating niosomal-PEG nanoparticles co-loaded with metformin and silibinin for effective treatment of human lung cancer cells. Front Oncol. 2023; 13:1193708.
5. Woźniak Ł, Skąpska S, Marszałek K. The synergistic effect of piperlongumine and sanguinarine on the non-small lung cancer (A549) cell line. Molecules. 2020;25(15):3383.
6. Suárez N, Rangel N, González-Reyes JA, et al. Paclitaxel and the dietary flavonoid fisetin: a synergistic combination that induces mitotic catastrophe and autophagic cell death in A549 non-small cell lung cancer cells. Cancer Cell Int. 2016; 16:24.
7. Panahi Y, Ghanei M, Bashiri S, Hajihashemi A, Sahebkar A. Enhanced anticancer effect of curcumin combined with piperine on A549 and NCI-H292 human lung cancer cell lines. Curr Mol Med. 2016;16(9):798-806.
8. Rahmani AH, Alsahli MA, Almatroodi SA. Synergistic induction of apoptosis by boswellic acid and cisplatin in A549 lung cancer cells through NF-κB modulation and p53 pathway activation. Molecules. 2024;29(20):4920.
9. Chou TC. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res. 2010;70(2):440-6.
10. Chou TC, Talalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul. 1984; 22:27-55.
11. Chou TC, Talalay P. Analysis of combined drug effects: a new look at a very old problem. Trends Pharmacol Sci. 1983; 4:450-4.
12. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65(1-2):55-63.
13. Berridge MV, Herst PM, Tan AS. Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction. Biotechnol Annu Rev. 2005; 11:127-52.
14. Sylvester PW. Optimization of the tetrazolium dye (MTT) colorimetric assay for cellular growth and viability. Methods Mol Biol. 2011; 716:157-68.
15. Kvasnicka F, Bíba B, Ševčík R, Voldřich M, Krátká J. Analysis of the active components of silymarin. J Chromatogr A. 2003;990(1-2):239-45.
16. Wianowska D, Wiśniewski M. Simplified procedure of silymarin extraction from Silybum marianum L. Gaertner. J Chromatogr Sci. 2015;53(2):366-72.
17. Duan L, Carrier DJ, Clausen EC. Extraction of silymarin from milk thistle (Silybum marianum) seeds - a comparison of conventional and microwave-assisted extraction methods. Sep Sci Technol. 2004;39(16):3831-42.
18. Abd El-Kader AM, Taha RA, Abou-Aisha K, El-Azizi M. Tackling of renal carcinogenesis in Wistar rats by Silybum marianum total extract, silymarin, and silibinin via modulation of oxidative stress, apoptosis, Nrf2, PPARγ, NF-κB, and PI3K/Akt signaling pathways. Oxid Med Cell Longev. 2021; 2021:9993869.
19. AbouZid S, Ahmed OM, Ahmed RR, Mahmoud A, Abdella E, Ashour MB. Silymarin and silibinin fraction from Silybum marianum barks: chromatographic isolation, HPLC identification, and hypoglycemic evaluation. Nat Prod Res. 2016;30(20):2350-4.
20. Bijak M. Silybin, a major bioactive component of milk thistle (Silybum marianum L. Gaernt.)-chemistry, bioavailability, and metabolism. Molecules. 2017;22(11):1942.

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