Skip to main content
Top
Published in: Cancer Cell International 1/2017

Open Access 01-12-2017 | Primary Research

Synergistic anticancer effect of combined crocetin and cisplatin on KYSE-150 cells via p53/p21 pathway

Authors: Sheng Li, Xiu-Yin Shen, Ting Ouyang, Yuhua Qu, Tao Luo, Hua-Qiao Wang

Published in: Cancer Cell International | Issue 1/2017

Login to get access

Abstract

Background

More than 400,000 patients die from esophageal cancer annually. Considerable efforts have been made to develop new and effective treatments, one of which is directed toward herbal medication. Crocetin is a natural carotenoid dicarboxylic acid isolated from the Chinese herb saffron. We recently reported on the anticancer effects of saffron. This study aimed to determine whether crocetin combined cisplatin has synergistic effect in KYSE-150 cells and explore the underlying mechanism.

Methods

KYSE-150 cells were treated with crocetin and/or cisplatin. The effects on cell viability, cell apoptosis, mitochondrial membrane potential (MMP), as well as the expression levels of PI3K/AKT, MAPKs, p53/p21, and apoptosis-related protein were evaluated. MTT assay, Annexin V-FITC/PI staining, Rh123 staining, and Western blot analysis were used.

Results

The cell proliferation significantly decreased and cell apoptosis was induced with combined crocetin and cisplatin, compared with either crocetin only or cisplatin only. The outcome suggested that crocetin combined cisplatin has synergistic effects on inhibition of cell proliferation and pro-apoptotic effect of cisplatin on KYSE-150 cells. Disruption of MMP, upregulation of cleaved caspase-3 expression, and downregulation of Bcl-2 occurred in the group treated with combined treatment. No significant differences in p-PI3K, p-AKT, and MAPKs activity were indicated between combined treatment group and the individual treatment group. However, the expression levels of p53 and p21 were markedly higher in the combined treatment group than in the individual treatment group. The wild-type p53 inhibitor, PFT-α suppressed the overexpression of p53/p21 and the synergistic effect induced by the combination of crocetin and cisplatin.

Conclusions

We concluded that crocetin combined with cisplatin exerts a synergistic anticancer effect by up-regulating the p53/p21 pathway.
Literature
1.
go back to reference Napier KJ, Scheerer M, Misra S. Esophageal cancer: a review of epidemiology, pathogenesis, staging workup and treatment modalities. World J Gastrointest Oncol. 2014;6(5):112–20.CrossRefPubMedPubMedCentral Napier KJ, Scheerer M, Misra S. Esophageal cancer: a review of epidemiology, pathogenesis, staging workup and treatment modalities. World J Gastrointest Oncol. 2014;6(5):112–20.CrossRefPubMedPubMedCentral
2.
go back to reference Kim T, Grobmyer SR, Smith R, Ben-David K, Ang D, Vogel SB, Hochwald SN. Esophageal cancer—the five year survivors. J Surg Oncol. 2011;103(2):179–83.CrossRefPubMed Kim T, Grobmyer SR, Smith R, Ben-David K, Ang D, Vogel SB, Hochwald SN. Esophageal cancer—the five year survivors. J Surg Oncol. 2011;103(2):179–83.CrossRefPubMed
3.
go back to reference Lisičić D, Benković V, Đikić D, Blažević AS, Mihaljević J, Oršolić N, Knežević AH. Addition of propolis to irinotecan therapy prolongs survival in ehrlich ascites tumor-bearing mice. Cancer Biother Radiopharm. 2014;29(2):62–9.CrossRefPubMedPubMedCentral Lisičić D, Benković V, Đikić D, Blažević AS, Mihaljević J, Oršolić N, Knežević AH. Addition of propolis to irinotecan therapy prolongs survival in ehrlich ascites tumor-bearing mice. Cancer Biother Radiopharm. 2014;29(2):62–9.CrossRefPubMedPubMedCentral
5.
go back to reference Biswal BM, Sulaiman SA, Ismail HC, Zakaria H, Musa KI. Effect of Withania somnifera (Ashwagandha) on the development of chemotherapy-induced fatigue and quality of life in breast cancer patients. Integr Cancer Ther. 2013;12(4):312–22.CrossRefPubMed Biswal BM, Sulaiman SA, Ismail HC, Zakaria H, Musa KI. Effect of Withania somnifera (Ashwagandha) on the development of chemotherapy-induced fatigue and quality of life in breast cancer patients. Integr Cancer Ther. 2013;12(4):312–22.CrossRefPubMed
6.
go back to reference Bolhassani A, Khavari A, Bathaie SZ. Saffron and natural carotenoids: biochemical activities and anti-tumor effects. Biochim Biophys Acta Rev Cancer. 2014;1845(1):20–30.CrossRef Bolhassani A, Khavari A, Bathaie SZ. Saffron and natural carotenoids: biochemical activities and anti-tumor effects. Biochim Biophys Acta Rev Cancer. 2014;1845(1):20–30.CrossRef
7.
go back to reference Nam KN, Park Y, Jung H, Lee JY, Min BD, Park S, Jung W, Cho K, Park J, Kang I, et al. Anti-inflammatory effects of crocin and crocetin in rat brain microglial cells. Eur J Pharmacol. 2010;648(1–3):110–6.CrossRefPubMed Nam KN, Park Y, Jung H, Lee JY, Min BD, Park S, Jung W, Cho K, Park J, Kang I, et al. Anti-inflammatory effects of crocin and crocetin in rat brain microglial cells. Eur J Pharmacol. 2010;648(1–3):110–6.CrossRefPubMed
8.
go back to reference Bathaie SZ, Hoshyar R, Miri H, Sadeghizadeh M. Anticancer effects of crocetin in both human adenocarcinoma gastric cancer cells and rat model of gastric cancer. Biochem Cell Biol. 2013;91(6):397–403.CrossRefPubMed Bathaie SZ, Hoshyar R, Miri H, Sadeghizadeh M. Anticancer effects of crocetin in both human adenocarcinoma gastric cancer cells and rat model of gastric cancer. Biochem Cell Biol. 2013;91(6):397–403.CrossRefPubMed
9.
go back to reference Li C, Huang W, Wang Q, Wang F, Cai E, Hu B, Du J, Wang J, Chen R, Cai X, et al. Crocetin induces cytotoxicity in colon cancer cells via p53-independent mechanisms. Asian Pac J Cancer Prev. 2012;13(8):3757–61.CrossRefPubMed Li C, Huang W, Wang Q, Wang F, Cai E, Hu B, Du J, Wang J, Chen R, Cai X, et al. Crocetin induces cytotoxicity in colon cancer cells via p53-independent mechanisms. Asian Pac J Cancer Prev. 2012;13(8):3757–61.CrossRefPubMed
10.
go back to reference Chen B, Hou ZH, Dong Z, Li CD. Crocetin downregulates the proinflammatory cytokines in methylcholanthrene-induced rodent tumor model and inhibits COX-2 expression in cervical cancer cells. Biomed Res Int. 2015;2015:829513.PubMedPubMedCentral Chen B, Hou ZH, Dong Z, Li CD. Crocetin downregulates the proinflammatory cytokines in methylcholanthrene-induced rodent tumor model and inhibits COX-2 expression in cervical cancer cells. Biomed Res Int. 2015;2015:829513.PubMedPubMedCentral
11.
go back to reference Li S, Jiang S, Jiang W, Zhou Y, Shen XY, Luo T, Kong LP, Wang HQ. Anticancer effects of crocetin in human esophageal squamous cell carcinoma KYSE-150 cells. Oncol Lett. 2015;9:1254–60.PubMedPubMedCentral Li S, Jiang S, Jiang W, Zhou Y, Shen XY, Luo T, Kong LP, Wang HQ. Anticancer effects of crocetin in human esophageal squamous cell carcinoma KYSE-150 cells. Oncol Lett. 2015;9:1254–60.PubMedPubMedCentral
12.
go back to reference Li X, Huang T, Jiang G, Gong W, Qian H, Zou C. Synergistic apoptotic effect of crocin and cisplatin on osteosarcoma cells via caspase induced apoptosis. Toxicol Lett. 2013;221(3):197–204.CrossRefPubMed Li X, Huang T, Jiang G, Gong W, Qian H, Zou C. Synergistic apoptotic effect of crocin and cisplatin on osteosarcoma cells via caspase induced apoptosis. Toxicol Lett. 2013;221(3):197–204.CrossRefPubMed
13.
go back to reference Shi H, Wu Y, Wang Y, Zhou M, Yan S, Chen Z, Gu D, Cai Y. Liquiritigenin potentiates the inhibitory effects of cisplatin on invasion and metastasis via downregulation MMP-2/9 and PI3K/AKT signaling pathway in B16F10 melanoma cells and mice model. Nutr Cancer. 2015;67:761–70.CrossRefPubMed Shi H, Wu Y, Wang Y, Zhou M, Yan S, Chen Z, Gu D, Cai Y. Liquiritigenin potentiates the inhibitory effects of cisplatin on invasion and metastasis via downregulation MMP-2/9 and PI3K/AKT signaling pathway in B16F10 melanoma cells and mice model. Nutr Cancer. 2015;67:761–70.CrossRefPubMed
14.
go back to reference 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.CrossRefPubMed 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.CrossRefPubMed
15.
go back to reference Zhang G, Gurtu V, Kain SR, Yan G. Early detection of apoptosis using a fluorescent conjugate of annexin V. Biotechniques. 1997;23(3):525–31.PubMed Zhang G, Gurtu V, Kain SR, Yan G. Early detection of apoptosis using a fluorescent conjugate of annexin V. Biotechniques. 1997;23(3):525–31.PubMed
16.
go back to reference Chen X, Wen C, Wu J, Su Y, Hong J. Giant seaperch iridovirus (GSIV) induces mitochondria-mediated cell death that is suppressed by bongkrekic acid and cycloheximide in a fish cell line. Virus Res. 2016;213:37–45.CrossRefPubMed Chen X, Wen C, Wu J, Su Y, Hong J. Giant seaperch iridovirus (GSIV) induces mitochondria-mediated cell death that is suppressed by bongkrekic acid and cycloheximide in a fish cell line. Virus Res. 2016;213:37–45.CrossRefPubMed
17.
go back to reference Gutheil WG, Reed G, Ray A, Anant S, Dhar A. Crocetin: an agent derived from saffron for prevention and therapy for cancer. Curr Pharm Biotechnol. 2012;13(1):173–9.CrossRefPubMedPubMedCentral Gutheil WG, Reed G, Ray A, Anant S, Dhar A. Crocetin: an agent derived from saffron for prevention and therapy for cancer. Curr Pharm Biotechnol. 2012;13(1):173–9.CrossRefPubMedPubMedCentral
18.
go back to reference Bathaie SZ, Farajzade A, Hoshyar R. A review of the chemistry and uses of crocins and crocetin, the carotenoid natural dyes in saffron, with particular emphasis on applications as colorants including their use as biological stains. Biotech Histochem. 2014;89(6):401–11.CrossRefPubMed Bathaie SZ, Farajzade A, Hoshyar R. A review of the chemistry and uses of crocins and crocetin, the carotenoid natural dyes in saffron, with particular emphasis on applications as colorants including their use as biological stains. Biotech Histochem. 2014;89(6):401–11.CrossRefPubMed
19.
go back to reference Chen J, Lan T, Zhang W, Dong L, Kang N, Zhang S, Fu M, Liu B, Liu K, Zhang C, et al. Platelet-activating factor receptor-mediated PI3K/AKT activation contributes to the malignant development of esophageal squamous cell carcinoma. Oncogene. 2015;34:5114–27.CrossRefPubMed Chen J, Lan T, Zhang W, Dong L, Kang N, Zhang S, Fu M, Liu B, Liu K, Zhang C, et al. Platelet-activating factor receptor-mediated PI3K/AKT activation contributes to the malignant development of esophageal squamous cell carcinoma. Oncogene. 2015;34:5114–27.CrossRefPubMed
20.
go back to reference Shigaki H, Baba Y, Watanabe M, Murata A, Ishimoto T, Iwatsuki M, Iwagami S, Nosho K, Baba H. PIK3CA mutation is associated with a favorable prognosis among patients with curatively resected esophageal squamous cell carcinoma. Clin Cancer Res. 2013;19(9):2451–9.CrossRefPubMed Shigaki H, Baba Y, Watanabe M, Murata A, Ishimoto T, Iwatsuki M, Iwagami S, Nosho K, Baba H. PIK3CA mutation is associated with a favorable prognosis among patients with curatively resected esophageal squamous cell carcinoma. Clin Cancer Res. 2013;19(9):2451–9.CrossRefPubMed
21.
go back to reference Li B, Li J, Xu WW, Guan XY, Qin YR, Zhang LY, Law S, Tsao SW, Cheung AL. Suppression of esophageal tumor growth and chemoresistance by directly targeting the PI3K/AKT pathway. Oncotarget. 2014;5(22):11576–87.CrossRefPubMedPubMedCentral Li B, Li J, Xu WW, Guan XY, Qin YR, Zhang LY, Law S, Tsao SW, Cheung AL. Suppression of esophageal tumor growth and chemoresistance by directly targeting the PI3K/AKT pathway. Oncotarget. 2014;5(22):11576–87.CrossRefPubMedPubMedCentral
22.
go back to reference Hildebrandt MA, Yang H, Hung MC, Izzo JG, Huang M, Lin J, Ajani JA, Wu X. Genetic variations in the PI3K/PTEN/AKT/mTOR pathway are associated with clinical outcomes in esophageal cancer patients treated with chemoradiotherapy. J Clin Oncol. 2009;27(6):857–71.CrossRefPubMedPubMedCentral Hildebrandt MA, Yang H, Hung MC, Izzo JG, Huang M, Lin J, Ajani JA, Wu X. Genetic variations in the PI3K/PTEN/AKT/mTOR pathway are associated with clinical outcomes in esophageal cancer patients treated with chemoradiotherapy. J Clin Oncol. 2009;27(6):857–71.CrossRefPubMedPubMedCentral
23.
go back to reference Siddik ZH. Cisplatin: mode of cytotoxic action and molecular basis of resistance. Oncogene. 2003;22(47):7265–79.CrossRefPubMed Siddik ZH. Cisplatin: mode of cytotoxic action and molecular basis of resistance. Oncogene. 2003;22(47):7265–79.CrossRefPubMed
24.
go back to reference Mebratu Y, Tesfaigzi Y. How ERK1/2 activation controls cell proliferation and cell death: is subcellular localization the answer? Cell Cycle. 2009;8(8):1168–75.CrossRefPubMedPubMedCentral Mebratu Y, Tesfaigzi Y. How ERK1/2 activation controls cell proliferation and cell death: is subcellular localization the answer? Cell Cycle. 2009;8(8):1168–75.CrossRefPubMedPubMedCentral
25.
go back to reference Recio JA, Merlino G. Hepatocyte growth factor/scatter factor activates proliferation in melanoma cells through p38 MAPK, ATF-2 and cyclin D1. Oncogene. 2002;21(7):1000–8.CrossRefPubMed Recio JA, Merlino G. Hepatocyte growth factor/scatter factor activates proliferation in melanoma cells through p38 MAPK, ATF-2 and cyclin D1. Oncogene. 2002;21(7):1000–8.CrossRefPubMed
26.
go back to reference Souza RF, Shewmake K, Terada LS, Spechler SJ. Acid exposure activates the mitogen-activated protein kinase pathways in Barrett’s esophagus. Gastroenterology. 2002;122:299–307.CrossRefPubMed Souza RF, Shewmake K, Terada LS, Spechler SJ. Acid exposure activates the mitogen-activated protein kinase pathways in Barrett’s esophagus. Gastroenterology. 2002;122:299–307.CrossRefPubMed
27.
go back to reference Cui X, Li S, Li T, Pang X, Zhang S, Jin J, Hu J, Liu C, Yang L, Peng H, et al. Significance of elevated ERK expression and its positive correlation with EGFR in Kazakh patients with esophageal squamous cell carcinoma. Int J Clin Exp Pathol. 2014;7(5):2382–91.PubMedPubMedCentral Cui X, Li S, Li T, Pang X, Zhang S, Jin J, Hu J, Liu C, Yang L, Peng H, et al. Significance of elevated ERK expression and its positive correlation with EGFR in Kazakh patients with esophageal squamous cell carcinoma. Int J Clin Exp Pathol. 2014;7(5):2382–91.PubMedPubMedCentral
28.
go back to reference Qin X, Zheng S, Liu T, Liu Q, Liang M, Li X, Sheyhidin I, Lu X. Roles of phosphorylated JNK in esophageal squamous cell carcinomas of Kazakh ethnic. Mol Carcinogen. 2014;53(7):526–36.CrossRef Qin X, Zheng S, Liu T, Liu Q, Liang M, Li X, Sheyhidin I, Lu X. Roles of phosphorylated JNK in esophageal squamous cell carcinomas of Kazakh ethnic. Mol Carcinogen. 2014;53(7):526–36.CrossRef
29.
go back to reference Zheng S, Zhang C, Qin X, Gen Y, Liu T, Sheyhidin I, Lu X. The status of phosphorylated p38 in esophageal squamous cell carcinoma. Mol Biol Rep. 2012;39(5):5315–21.CrossRefPubMed Zheng S, Zhang C, Qin X, Gen Y, Liu T, Sheyhidin I, Lu X. The status of phosphorylated p38 in esophageal squamous cell carcinoma. Mol Biol Rep. 2012;39(5):5315–21.CrossRefPubMed
30.
go back to reference Zhai H, Hu S, Liu T, Wang F, Wang X, Wu G, Zhang Y, Sui M, Liu H, Jiang L. Nitidine chloride inhibits proliferation and induces apoptosis in colorectal cancer cells by suppressing the ERK signaling pathway. Mol Med Rep. 2016;13(3):2536–42.CrossRefPubMedPubMedCentral Zhai H, Hu S, Liu T, Wang F, Wang X, Wu G, Zhang Y, Sui M, Liu H, Jiang L. Nitidine chloride inhibits proliferation and induces apoptosis in colorectal cancer cells by suppressing the ERK signaling pathway. Mol Med Rep. 2016;13(3):2536–42.CrossRefPubMedPubMedCentral
31.
go back to reference Fu X, Yang Y, Li X, Lai H, Huang Y, He L, Zheng W, Chen T. RGD peptide-conjugated selenium nanoparticles: antiangiogenesis by suppressing VEGF-VEGFR2-ERK/AKT pathway. Nanomedicine. 2016;12:1627–39.CrossRefPubMed Fu X, Yang Y, Li X, Lai H, Huang Y, He L, Zheng W, Chen T. RGD peptide-conjugated selenium nanoparticles: antiangiogenesis by suppressing VEGF-VEGFR2-ERK/AKT pathway. Nanomedicine. 2016;12:1627–39.CrossRefPubMed
32.
go back to reference Shi X, Wu S, Yang Y, Tang L, Wang Y, Dong J, Lu B, Jiang G, Zhao W. AQP5 silencing suppresses p38 MAPK signaling and improves drug resistance in colon cancer cells. Tumour Biol. 2014;35(7):7035–45.CrossRefPubMed Shi X, Wu S, Yang Y, Tang L, Wang Y, Dong J, Lu B, Jiang G, Zhao W. AQP5 silencing suppresses p38 MAPK signaling and improves drug resistance in colon cancer cells. Tumour Biol. 2014;35(7):7035–45.CrossRefPubMed
33.
go back to reference Seta T, Imazeki F, Yokosuka O, Saisho H, Suzuki T, Koide Y, Isono K. Expression of p53 and p21WAF1/CIP1 proteins in gastric and esophageal cancers: comparison with mutations of the p53 gene. Dig Dis Sci. 1998;43(2):279–89.CrossRefPubMed Seta T, Imazeki F, Yokosuka O, Saisho H, Suzuki T, Koide Y, Isono K. Expression of p53 and p21WAF1/CIP1 proteins in gastric and esophageal cancers: comparison with mutations of the p53 gene. Dig Dis Sci. 1998;43(2):279–89.CrossRefPubMed
Metadata
Title
Synergistic anticancer effect of combined crocetin and cisplatin on KYSE-150 cells via p53/p21 pathway
Authors
Sheng Li
Xiu-Yin Shen
Ting Ouyang
Yuhua Qu
Tao Luo
Hua-Qiao Wang
Publication date
01-12-2017
Publisher
BioMed Central
Published in
Cancer Cell International / Issue 1/2017
Electronic ISSN: 1475-2867
DOI
https://doi.org/10.1186/s12935-017-0468-9

Other articles of this Issue 1/2017

Cancer Cell International 1/2017 Go to the issue
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine