Original Article

The Synergistic Combination of Cisplatin and Piperine Induces Apoptosis in MCF-7 Cell Line


Background: Piperine is a natural compound obtained from the Piper nigrum that exhibits anti-proliferative and anti-cancer activity in cancer cell lines. We analyzed the cytotoxic effect of piperine combined with cisplatin compound in the human MCF-7 breast cancer cell line and the underlying mechanism.

Methods: The present in vitro study was performed on MCF-7 cell line in Jahrom University of Medical Sciences between, Jahrom, Iran from 2016 to 2017. Cultured MCF-7 cells were seeded into four groups: a control group (untreated group), a group treated with cisplatin, a group treated with piperine and a group treated with cisplatin and piperine. Cell viability was analyzed using the MTT assay method. Flow c-ytometric analysis was investigated for apoptosis. The mRNA and protein expression of the apoptotic regulators p53, Bcl-2, Bax, caspase 3 and caspase 9 were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting analysis.

Results: Piperine (20 and 30 µM) in combination with cisplatin (5, 10 and 15 µM) for 24 h synergistically inhibited cell viability of MCF-7 breast cancer cells more than piperine and cisplatin used alone. Synergistic anti-breast cancer activities cisplatin (5 µM) and piperine (20 µM) were via inducing apoptosis. Piperine (20 µM) and cisplatin (5 µM) for 24 h induce apoptosis strongly through reduction of Bcl-2 and increase of caspase 3, p53, caspase 9, and Bax.

Conclusion: Piperine in combination with cisplatin could trigger p53-mediated apoptosis more effective than cisplatin alone in MCF-7 breast cancer cells, reducing the toxic dose of cisplatin used in cancer chemotherapy.

1. DeSantis CE, Ma J, Goding Sauer A, et al (2017). Breast cancer statistics, 2017, racial disparity in mortality by state. CA Cancer J Clin, 67(6):439-448.
2. Florea A-M, Büsselberg D (2011). Cisplatin as an anti-tumor drug: cellular mechanisms of activity, drug resistance and induced side effects. Cancers (Basel), 3(1):1351-71.
3. Abazari O, Divsalar A, Ghobadi R (2019). Inhibitory effects of oxali-Platin as a chemotherapeutic drug on the function and structure of bovine liver catalase. J Biomol Struct Dyn, 38(2):609-615.
4. Basu A, Krishnamurthy S (2010). Cellular responses to Cisplatin-induced DNA damage. J Nucleic Acids, 2010: 201367.
5. Petrović M, Todorović D (2016). Biochemical and molecular mechanisms of action of cisplatin in cancer cells. Facta Univ Ser Med Biol, 12-18.
6. Asadi A, Nezhad DY, Javazm AR, et al (2020). In Vitro Effects of Curcumin on Transforming Growth Factor-β-mediated Non-Smad Signaling Pathway, Oxidative Stress, and Pro‐inflammatory Cytokines Production with Human Vascular Smooth Muscle Cells. Iran J Allergy Asthma Immunol, 19(1):84-93.
7. Cepeda V, Fuertes MA, Castilla J, et al (2007). Biochemical mechanisms of cisplatin cytotoxicity. Anticancer Agents Med Chem, 7(1):3-18.
8. Oun R, Moussa YE, Wheate NJ (2018). The side effects of platinum-based chemotherapy drugs: a review for chemists. Dalton Trans, 47(19):6645-6653.
9. Gordaliza M (2007). Natural products as leads to anticancer drugs. Clin Transl Oncol, 9(12):767-76.
10. Abazari O, Divsalar A, Ghobadi R (2020). Inhibitory effects of oxali-Platin as a chemotherapeutic drug on the function and structure of bovine liver catalase. J Biomol Struct Dyn, 38(2):609-615.
11. Sharifat N, Jafari-Hafshejani F, Dayati P, et al (2017). Inhibitory effect of Curcumin on phosphorylation NFκB-p65 induced by hydrogen peroxide in Bovine Endothelial Cells. J Fasa Univ Med Sci, 7(2): 283-290.
12. Rather RA, Bhagat M (2018). Cancer Chemoprevention and Piperine: Molecular Mechanisms and Therapeutic Opportunities. Front Cell Dev Biol, 6:10.
13. Zuzanna Bober Z, Agnieszka Stępień A, David Aebisher D, et al (2018). Medicinal benefits from the use of Black pepper, Curcuma and Ginger. Eur J Clin Exp Med:133-145.
14. Dayati P, Rezaei HB, Sharifat N, et al (2018). G protein coupled receptors can transduce signals through carboxy terminal and linker region phosphorylation of Smad transcription factors. Life sci, 199:10-15.
15. Deng Y, Sriwiriyajan S, Tedasen A, et al (2016). Anti-cancer effects of Piper nigrum via inducing multiple molecular signaling in vivo and in vitro. J Ethnopharmacol, 188:87-95.
16. Lin Y, Xu J, Liao H, et al (2014). Piperine induces apoptosis of lung cancer A549 cells via p53-dependent mitochondrial signaling pathway. Tumour Biol, 35(4):3305-10.
17. Yaffe PB, Power Coombs MR, Doucette CD, et al (2015). Piperine, an alkaloid from black pepper, inhibits growth of human colon cancer cells via G1 arrest and apoptosis triggered by endoplasmic reticulum stress. Mol Carcinog, 54(10):1070-85.
18. Abazari O, Shafaei Z, Divsalar A, et al (2016). Probing the biological evaluations of a new designed Pt (II) complex using spectroscopic and theoretical approaches: Human hemoglobin as a target. J Biomol Struct Dyn, 34(5):1123-31.
19. Sayers TJ (2011). Targeting the extrinsic apoptosis signaling pathway for cancer therapy. Cancer Immunol Immunother, 60(8):1173-80.
20. Han S-z, Liu H-x, Yang L-q, et al (2017). Piperine (PP) enhanced mitomycin-C (MMC) therapy of human cervical cancer through suppressing Bcl-2 signaling pathway via inactivating STAT3/NF-κB. Biomed Pharmacother, 96:1403-1410.
21. Abazari O, Shafaei Z, Divsalar A, et al (2020). Interaction of the synthesized anticancer compound of the methyl-glycine 1, 10-phenanthroline platinum nitrate with human serum albumin and human hemoglobin proteins by spectroscopy methods and molecular docking. Journal of the Iranian Chemical Society, 17:1601-1614.
22. Chou T-C, Talalay P (1984). Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Advances in Biological Regulation, 22:27-55.
23. Livak KJ, Schmittgen TD (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 25(4):402-8.
24. Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 72:248-54.
25. Dasari S, Tchounwou PB (2014). Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol, 740:364-78.
26. Desoize B, Madoulet C (2002). Particular aspects of platinum compounds used at present in cancer treatment. Crit Rev Oncol Hematol, 42(3):317-25.
27. Musavi H, Abazari O, Barartabar Z, et al (2020). The benefits of Vitamin D in the COVID-19 pandemic: biochemical and immunological mechanisms. Arch Physiol Biochem,1-9.
28. Mohamed R, Dayati P, Mehr RN, et al (2019). Transforming growth factor–β1 mediated CHST11 and CHSY1 mRNA expression is ROS dependent in vascular smooth muscle cells. J Cell Commun Signal, 13(2):225-233.
29. Galluzzi L, Senovilla L, Vitale I, et al (2012). Molecular mechanisms of cisplatin resistance. Oncogene, 31(15):1869-83.
30. Demain AL, Vaishnav P (2011). Natural products for cancer chemotherapy. Microb Biotechnol, 4(6): 687–699.
31. Nobili S, Lippi D, Witort E, et al (2009). Natural compounds for cancer treatment and prevention. Pharmacol Res, 59(6):365-78.
32. Si L, Yang R, Lin R, et al (2018). Piperine functions as a tumor suppressor for human ovarian tumor growth via activation of JNK/p38 MAPK-mediated intrinsic apoptotic pathway. Biosci Rep, 38(3):BSR20180503.
33. Lai L-h, Fu Q-h, Liu Y, et al (2012). Piperine suppresses tumor growth and metastasis in vitro and in vivo in a 4T1 murine breast cancer model. Acta Pharmacol Sin, 33(4): 523–530.
34. Greenshields AL, Doucette CD, Sutton KM, et al (2015). Piperine inhibits the growth and motility of triple-negative breast cancer cells. Cancer Lett, 357(1):129-140.
35. Pal MK, Jaiswar SP, Srivastav AK, et al (2016). Synergistic effect of piperine and paclitaxel on cell fate via cyt-c, Bax/Bcl-2-caspase-3 pathway in ovarian adenocarcinomas SKOV-3 cells. Eur J Pharmacol, 791:751-762.
36. Musavi H, Abazari O, Safaee MS, et al (2021). Mechanisms of COVID-19 Entry into the Cell: Potential Therapeutic Approaches Based on Virus Entry Inhibition in COVID-19 Patients with Underlying Diseases. Iran J Allergy Asthma Immunol, 20(1):11-23.
37. Abbasi M, Abazari OO (2018). Probing the Biological evaluations of a new designed Palladium (II) complex using spectroscopic and theoretical approaches: Human Hemoglobin as a Target. Archives of Medical Laboratory Sciences, doi.org/10.22037/amls.v3i3.21712.
38. Czabotar PE, Lessene G, Strasser A, et al (2014). Control of apoptosis by the BCL-2 protein family: implications for physiology and therapy. Nat Rev Mol Cell Biol, 15(1):49-63.
39. Sharifat N, Mohammad Zadeh G, Ghaffari MA, et al (2017). Endothelin‐1 (ET‐1) stimulates carboxy terminal Smad2 phosphorylation in vascular endothelial cells by a mechanism dependent on ET receptors and de novo protein synthesis. J Pharm Pharmacol, 69(1):66-72.
40. Zare Z, Dizaj TN, Lohrasbi A, et al (2020). Silibinin inhibits TGF-β-induced MMP-2 and MMP-9 through Smad Signaling pathway in colorectal cancer HT-29 cells. Basic & Clinical Cancer Research, 12(2): 81-90.
41. Sharifi S, Barar J, Hejazi MS, et al (2015). Doxorubicin changes Bax/Bcl-xL ratio, caspase-8 and 9 in breast cancer cells. Adv Pharm Bull, 5(3): 351–359.
42. Zare Z, Dizaj TN, Lohrasbi A, et al (2020). The Effect of Piperine on MMP-9, VEGF, and E-cadherin Expression in Breast Cancer MCF-7 Cell Line. Basic & Clinical Cancer Research, 12(3):112-119.
43. Kim B, Srivastava SK, Kim S-H (2015). Caspase-9 as a therapeutic target for treating cancer. Expert Opin Ther Targets, 19(1):113-27.
IssueVol 50 No 5 (2021) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijph.v50i5.6121
Apoptosis Breast cancer Caspase Cisplatin Piperine

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
Fattah A, Morovati A, Niknam Z, Mashouri L, Asadi A, Tvangar Rizi S, Abbasi M, Shakeri F, Abazari O. The Synergistic Combination of Cisplatin and Piperine Induces Apoptosis in MCF-7 Cell Line. Iran J Public Health. 50(5):1037-1047.