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Melatonin induces apoptosis and cell cycle arrest in cervical cancer cells via inhibition of NF-κB pathway

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Abstract

Cervical cancer is the most prevalent cancer in females. Melatonin, a neurohormone has been documented as a promising therapeutic molecule for cervical cancer. However, the underlying molecular mechanism is not known. We explored the dose-dependent anti-tumor response of melatonin against cervical cancer cell lines, HeLa (HPV-18 positive) and SiHa (HPV-16 positive). The anti-cancer effect of melatonin was evaluated by MTT assay, cell imaging, colony formation, DAPI, AO/PI, LDH, Flow cytometry, scratch assay, western blot analysis and real-time PCR. Results of DAPI, AO/PI, LDH, and Annexin/PI staining revealed that melatonin induces apoptosis. The results of cell cycle analysis revealed that melatonin arrests the HeLa and SiHa cells in sub-G1 and G1 phases, respectively. Western blot analysis revealed that melatonin downregulated the expression of pro-inflammatory transcription factor, NF-κB and the expression of COX-2 protein, a key mediator in cell proliferation. In addition, melatonin downregulated the expression of an invasive marker, MMP-9, an antiapoptotic protein, Bcl-2, and upregulated the expression of pro-apoptotic protein, Bax at both transcriptional and translational levels. Overall, the results suggest that melatonin exhibited strong anti-cancer therapeutic potential against human cervical cancer cell line progression possibly through inhibition of NF-κB signalling pathway.

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References

  • Ahmad R, Haldar C (2010) Photoperiodic regulation of MT1 and MT2 melatonin receptor expression in spleen and thymus of a tropical rodent Funambulus pennanti during reproductively active and inactive phases. Chronobiol Int 27(3):446–462

    Article  CAS  PubMed  Google Scholar 

  • Altun I, Sonkaya A (2018) The most common side effects experienced by patients were receiving first cycle of chemotherapy. Iran J Public Health 47(8):1218–1219

    PubMed  PubMed Central  Google Scholar 

  • Asting AG, Carén H, Andersson M, Lönnroth C, Lagerstedt K, Lundholm K (2011) COX-2 gene expression in colon cancer tissue related to regulating factors and promoter methylation status. BMC Cancer 11(1):1–10

    Article  CAS  Google Scholar 

  • Awasthee N, Rai V, Verma SS, Francis KS, Nair MS, Gupta SC (2018) Anti-cancer activities of Bharangin against breast cancer: evidence for the role of NF-κB and lncRNAs. Biochim Et Biophys Acta (BBA) 1862(12):2738–2749

    Article  CAS  Google Scholar 

  • Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44(1):276–287

    Article  CAS  PubMed  Google Scholar 

  • Bentivegna E, Gouy S, Maulard A, Chargari C, Leary A, Morice P (2016) Oncological outcomes after fertility-sparing surgery for cervical cancer: a systematic review. Lancet Oncol 17(6):e240–e253

    Article  PubMed  Google Scholar 

  • Bikfalvi A (2017) History and conceptual developments in vascular biology and angiogenesis research: a personal view. Angiogenesis 20(4):463–478

    Article  PubMed  Google Scholar 

  • Burdon R (1996) Control of cell proliferation by reactive oxygen species. Biochem Soc Trans 24(4):1028–1032

    Article  CAS  PubMed  Google Scholar 

  • Cabrera J, Negrín G, Estévez F, Loro J, Reiter RJ, Quintana J (2010) Melatonin decreases cell proliferation and induces melanogenesis in human melanoma SK-MEL-1 cells. J Pineal Res 49(1):45–54

    CAS  PubMed  Google Scholar 

  • Chen G, Li X, Yang J, Li J, Wang X, He J et al (2016) Prognostic significance of cyclooxygenase-2 expression in patients with hepatocellular carcinoma: a meta-analysis. Arch Med Sci 12(5):1110

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Choi B-Y, Kim H-Y, Lee K-H, Cho Y-H, Kong G (1999) Clofilium, a potassium channel blocker, induces apoptosis of human promyelocytic leukemia (HL-60) cells via Bcl-2-insensitive activation of caspase-3. Cancer Lett 147(1–2):85–93

    Article  CAS  PubMed  Google Scholar 

  • Chuffa LGA, Fioruci-Fontanelli BA, Mendes LO, Seiva FRF, Martinez M, Fávaro WJ et al (2015) Melatonin attenuates the TLR4-mediated inflammatory response through MyD88-and TRIF-dependent signaling pathways in an in vivo model of ovarian cancer. BMC Cancer 15(1):1–13

    Article  CAS  Google Scholar 

  • Cury-Boaventura MF, Pompéia C, Curi R (2004) Comparative toxicity of oleic acid and linoleic acid on Jurkat cells. Clin Nutr 23(4):721–732

    Article  CAS  PubMed  Google Scholar 

  • da Costa RMG, Bastos MM, Medeiros R, Oliveira PA (2016) The NFκB signaling pathway in papillomavirus-induced lesions: friend or foe? Anticancer Res 36(5):2073–2083

    Google Scholar 

  • Festa-Vasconcellos JS, Piranda DN, Amaral LM, Indio-do-Brasil V, Koifman S, Vianna-Jorge R (2012) Polymorphisms in cycloxygenase-2 gene and breast cancer prognosis: association between PTGS2 haplotypes and histopathological features. Breast Cancer Res Treat 132(1):251–258

    Article  CAS  PubMed  Google Scholar 

  • Gately S, Li WW (2004) Multiple roles of COX-2 in tumor angiogenesis: a target for antiangiogenic therapy. Semin Oncol 31:2–11

    Article  CAS  PubMed  Google Scholar 

  • Ghosh N, Chaki R, Mandal V, Mandal SC (2010) COX-2 as a target for cancer chemotherapy. Pharmacol Rep 62(2):233–244

    Article  CAS  PubMed  Google Scholar 

  • Gomes M, Teixeira AL, Coelho A, Araújo A, Medeiros R (2014) The role of inflammation in lung cancer. Inflammation and cancer. Springer, Berlin, pp 1–23

    Google Scholar 

  • Gupta SC, Kim JH, Prasad S, Aggarwal BB (2010) Regulation of survival, proliferation, invasion, angiogenesis, and metastasis of tumor cells through modulation of inflammatory pathways by nutraceuticals. Cancer Metastasis Rev 29(3):405–434

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gupta SC, Prasad S, Sethumadhavan DR, Nair MS, Mo Y-Y, Aggarwal BB (2013) Nimbolide, a limonoid triterpene, inhibits growth of human colorectal cancer xenografts by suppressing the proinflammatory microenvironment. Clin Cancer Res 19(16):4465–4476

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hartwell LH, Kastan MB (1994) Cell cycle control and cancer. Science 266(5192):1821–1828

    Article  CAS  PubMed  Google Scholar 

  • Kwan YP, Saito T, Ibrahim D, Al-Hassan FMS, Ein Oon C, Chen Y et al (2016) Evaluation of the cytotoxicity, cell-cycle arrest, and apoptotic induction by Euphorbia hirta in MCF-7 breast cancer cells. Pharm Biol 54(7):1223–1236

    CAS  PubMed  Google Scholar 

  • Lai JC-Y, Chou Y-J, Huang N, Tsai J-J, Huang S-M, Yang Y-C et al (2013) Survival analysis of Stage IIA1 and IIA2 cervical cancer patients. Taiwan J Obstet Gynecol 52(1):33–38

    Article  PubMed  Google Scholar 

  • Li Y, Li S, Zhou Y, Meng X, Zhang J-J, Xu D-P et al (2017) Melatonin for the prevention and treatment of cancer. Oncotarget 8(24):39896

    Article  PubMed  PubMed Central  Google Scholar 

  • Liang X-J, Chen C, Zhao Y, Wang PC (2010) Circumventing tumor resistance to chemotherapy by nanotechnology. Multi-drug resistance in cancer. Springer, Berlin, pp 467–488

    Chapter  Google Scholar 

  • Lin CL, Chen HJ, Hou WC (2002) Activity staining of glutathione peroxidase after electrophoresis on native and sodium dodecyl sulfate polyacrylamide gels. Electrophoresis 23(4):513–516

    Article  CAS  PubMed  Google Scholar 

  • Lin CY, Lovén J, Rahl PB, Paranal RM, Burge CB, Bradner JE et al (2012) Transcriptional amplification in tumor cells with elevated c-Myc. Cell 151(1):56–67

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Muñoz N, Castellsagué X, de González AB, Gissmann L (2006) HPV in the etiology of human cancer. Vaccine 24:S1–S10

    Article  CAS  Google Scholar 

  • Okazawa M, Mabuchi S, Isohashi F, Suzuki O, Yoshioka Y, Sasano T et al (2013) Impact of the addition of concurrent chemotherapy to pelvic radiotherapy in surgically treated stage IB1-IIB cervical cancer patients with intermediate-risk or high-risk factors: a 13-year experience. Int J Gynecol Cancer 23(3):567–575

    Article  PubMed  Google Scholar 

  • Park E-J, Cheenpracha S, Chang LC, Kondratyuk TP, Pezzuto JM (2011) Inhibition of lipopolysaccharide-induced cyclooxygenase-2 and inducible nitric oxide synthase expression by 4-[(2′-O-acetyl-α-L-rhamnosyloxy) benzyl] isothiocyanate from Moringa oleifera. Nutr Cancer 63(6):971–982

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pittayapruek P, Meephansan J, Prapapan O, Komine M, Ohtsuki M (2016) Role of matrix metalloproteinases in photoaging and photocarcinogenesis. Int J Mol Sci 17(6):868

    Article  PubMed Central  CAS  Google Scholar 

  • Qin S, Zhang Z, Li J, Zang L (2014) FRZB knockdown upregulates β-catenin activity and enhances cell aggressiveness in gastric cancer. Oncol Rep 31(5):2351–2357

    Article  CAS  PubMed  Google Scholar 

  • Roa JC, Garcia P, Gomez J, Fernández W, Gaete F, Espinoza A et al (2009) HPV genotyping from invasive cervical cancer in Chile. Int J Gynecol Obstet 105(2):150–153

    Article  Google Scholar 

  • Rouhollahi E, Moghadamtousi SZ, Paydar M, Fadaeinasab M, Zahedifard M, Hajrezaie M et al (2015) Inhibitory effect of Curcuma purpurascens BI. rhizome on HT-29 colon cancer cells through mitochondrial-dependent apoptosis pathway. BMC Complement Altern Med 15(1):1–12

    Article  Google Scholar 

  • Saavedra KP, Brebi PM, Roa JCS (2012) Epigenetic alterations in preneoplastic and neoplastic lesions of the cervix. Clin Epigenetics 4(1):1–7

    Article  CAS  Google Scholar 

  • Savvidis C, Koutsilieris M (2012) Circadian rhythm disruption in cancer biology. Mol Med 18(9):1249–1260

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schwartzman RA, Cidlowski JA (1993) Apoptosis: the biochemistry and molecular biology of programmed cell death. Endocr Rev 14(2):133–151

    CAS  PubMed  Google Scholar 

  • Sharma G, Rana NK, Singh P, Dubey P, Pandey DS, Koch B (2017) p53 dependent apoptosis and cell cycle delay induced by heteroleptic complexes in human cervical cancer cells. Biomed Pharmacother 88:218–231

    Article  CAS  PubMed  Google Scholar 

  • Shiu SY, Law IC, Lau KW, Tam PC, Yip AW, Ng WT (2003) Melatonin slowed the early biochemical progression of hormone-refractory prostate cancer in a patient whose prostate tumor tissue expressed MT1 receptor subtype. J Pineal Res 35(3):177–182

    Article  CAS  PubMed  Google Scholar 

  • Sovak MA, Bellas RE, Kim DW, Zanieski GJ, Rogers AE, Traish AM et al (1997) Aberrant nuclear factor-kappaB/Rel expression and the pathogenesis of breast cancer. J Clin Investig 100(12):2952–2960

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Talib WH, Alsayed AR, Abuawad A, Daoud S, Mahmod AI (2021) Melatonin in cancer treatment: current knowledge and future opportunities. Molecules 26(9):2506

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Verma SS, Rai V, Awasthee N, Dhasmana A, Rajalaksmi DS, Nair MS et al (2019) Isodeoxyelephantopin, a sesquiterpene lactone induces ROS generation, suppresses NF-κB activation, modulates LncRNA expression and exhibit activities against breast cancer. Sci Rep 9(1):1–16

    Article  CAS  Google Scholar 

  • Wang T-H, Lin Y-S, Chen Y, Yeh C-T, Huang Y-L, Hsieh T-H et al (2015) Long non-coding RNA AOC4P suppresses hepatocellular carcinoma metastasis by enhancing vimentin degradation and inhibiting epithelial-mesenchymal transition. Oncotarget 6(27):23342

    Article  PubMed  PubMed Central  Google Scholar 

  • Wang T-H, Wu C-H, Yeh C-T, Su S-C, Hsia S-M, Liang K-H et al (2017) Melatonin suppresses hepatocellular carcinoma progression via lncRNA-CPS1-IT-mediated HIF-1α inactivation. Oncotarget 8(47):82280

    Article  PubMed  PubMed Central  Google Scholar 

  • Weydert CJ, Cullen JJ (2010) Measurement of superoxide dismutase, catalase and glutathione peroxidase in cultured cells and tissue. Nat Protoc 5(1):51–66

    Article  CAS  PubMed  Google Scholar 

  • Woodbury W, Spencer A, Stahmann M (1971) An improved procedure using ferricyanide for detecting catalase isozymes. Anal Biochem 44(1):301–305

    Article  CAS  PubMed  Google Scholar 

  • Yamaguchi H, Condeelis J (2007) Regulation of the actin cytoskeleton in cancer cell migration and invasion. Biochim Et Biophys Acta (BBA) 1773(5):642–652

    Article  CAS  Google Scholar 

  • Yang M, Huang C-Z (2015) Mitogen-activated protein kinase signaling pathway and invasion and metastasis of gastric cancer. World J Gastroenterol 21(41):11673

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yordanov G, Skrobanska R, Petkova M (2016) Poly (butyl cyanoacrylate) nanoparticles stabilised with poloxamer 188: particle size control and cytotoxic effects in cervical carcinoma (HeLa) cells. Chem Pap 70(3):365–374

    Article  CAS  Google Scholar 

  • Zhang HM, Zhang Y (2014) Melatonin: a well-documented antioxidant with conditional pro-oxidant actions. J Pineal Res 57(2):131–146

    Article  CAS  PubMed  Google Scholar 

  • Zhao Y-R, Li H-M, Zhu M, Li J, Ma T, Huo Q et al (2018) Non-benzoquinone geldanamycin analog, WK-88-1, induces apoptosis in human breast cancer cell lines. J Microbiol Biotechnol 28(4):542–550

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors are also thankful to Dr. Rahul Kumar Singh, Department of Zoology, BHU for providing Chemi-Doc facility. We are also thankful to Interdisciplinary School of Life Sciences (ISLS), BHU for flow cytometry and real-time PCR facilities.

Funding

The authors are thankful to the Indian Council of Medical Research (ICMR), New Delhi (ICMR Adhoc Project P-14/267) for the financial assistance to Dr. Sanjeev Kumar Yadav. The JRF fellowship to Mr. Tarun Minocha from UGC-CAS, Dept of Zoology, BHU is highly acknowledged.

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Authors and Affiliations

  1. Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi, 221005, India

    Tarun Minocha, Megha Das, Chandana Haldar & Sanjeev Kumar Yadav

  2. Department of Biochemistry, Institute of Science, Banaras Hindu University, Varanasi, 221005, India

    Vipin Rai, Sumit Singh Verma, Nikee Awasthee & Subash Chandra Gupta

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  1. Tarun Minocha
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  2. Megha Das
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Contributions

SKY contributed to the study design. TM and MD performed experiments. SSV, NA and VR performed the statistical analysis. SKY wrote the manuscript. SCG and CH revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to Sanjeev Kumar Yadav.

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Minocha, T., Das, M., Rai, V. et al. Melatonin induces apoptosis and cell cycle arrest in cervical cancer cells via inhibition of NF-κB pathway. Inflammopharmacol 30, 1411–1429 (2022). https://doi.org/10.1007/s10787-022-00964-6

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