Abstract
Aim
This research aims to explore the effect of alpinumisoflavone (AIF) as an anti-cancer drug for the treatment of patients with hepatocellular carcinoma (HCC).
Methods
Cell counting kit-8 (CCK-8) and colony formation assay were used to evaluate the viability of the cells and their clonogenic ability. Cellular migration and their invasion capabilities were detected using the wound-healing and transwell assay, respectively. The release of lactate dehydrogenase (LDH) was detected using the LDH kit. The expression levels of genes in the cells and tumor tissues were examined by qRT-PCR, western blotting, and immunohistochemical techniques. The cells transfected with mRFP-GFP-LC3 adenoviruses were stained to determine their autophagy status. MCC950 (NLRP3 inflammasome inhibitor) and NLRP3 shRNA were used to block NLRP3-mediated pyroptosis. Chloroquine and Atg 5 siRNA were used to inhibit the autophagy of the cells.
Results
AIF suppressed cell proliferation, migration, and invasion capacity of SMMC 7721 and Huh7 cells. The incorporation of AIF induced the formation of NLRP3 inflammasome assembly, pyroptosis, and autophagy of the cells. However, the anti-proliferative and anti-metastatic effects of AIF on the HCC cells were attenuated by NLRP3 inhibitor and knockdown. Furthermore, Atg 5 knockdown inhibited autophagy and enhanced the rate of AIF-induced pyroptosis of the cells. AIF also suppressed tumor growth and increased the levels of pyroptosis-related genes in tumor tissues, which were consistent with in vitro observations.
Conclusion
AIF inhibited HCC cell growth and metastasis by inducing NLRP3 inflammasome-mediated pyroptosis. Furthermore, AIF-induced autophagy augmented pyroptosis in HCC.
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References
Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68:394–424.
Llovet JM, Villanueva A, Lachenmayer A, Finn RS. Advances in targeted therapies for hepatocellular carcinoma in the genomic era. Nature Rev Clin Oncol. 2015;12:408–24.
Ahmed F, Ishibashi M. Bio-active natural products with TRAIL-resistance overcoming activity. Chem Pharm Bull. 2016;64:119–27.
Lyddiard JR, Whitfield PJ. Inhibition of site I mitochondrial electron transport by an extract of the seeds of Millettia thonningii: a potential mechanism for the plant’s molluscicidal and schistosome larvicidal activity. J Helminthol. 2001;75:259–65.
Mvondo MA, Njamen D, Tanee Fomum S, Wandji J. Effects of alpinumisoflavone and abyssinone V-4'-methyl ether derived from Erythrina lysistemon (Fabaceae) on the genital tract of ovariectomized female Wistar rat. Phytotherapy Res PTR. 2012;26:1029–36.
Chukwujekwu JC, Van Heerden FR, Van Staden J. Antibacterial activity of flavonoids from the stem bark of Erythrina caffra thunb. Phytother Res. 2011;25:46–8.
Wang T, Jiang Y, Chu L, Wu T, You J. Alpinumisoflavone suppresses tumour growth and metastasis of clear-cell renal cell carcinoma. Am J Cancer Res. 2017;7:999–1015.
Han Y, Yang X, Zhao N, Peng J, Gao H, Qiu X. Alpinumisoflavone induces apoptosis in esophageal squamous cell carcinoma by modulating miR-370/PIM1 signaling. Am J Cancer Res. 2016;6:2755–71.
Namkoong S, Kim TJ, Jang IS, Kang KW, Oh WK, Park J. Alpinumisoflavone induces apoptosis and suppresses extracellular signal-regulated kinases/mitogen activated protein kinase and nuclear factor-kappaB pathways in lung tumor cells. Biol Pharm Bull. 2011;34:203–8.
Li D, Li X, Li G, Meng Y, Jin Y, Shang S, et al. Alpinumisoflavone causes DNA damage in colorectal cancer cells via blocking DNA repair mediated by RAD51. Life Sci. 2019;216:259–70.
Zhang B, Fan X, Wang Z, Zhu W, Li J. Alpinumisoflavone radiosensitizes esophageal squamous cell carcinoma through inducing apoptosis and cell cycle arrest. Biomed Pharmacotherapy. 2017;95:199–206.
Bergsbaken T, Fink SL, Cookson BT. Pyroptosis: host cell death and inflammation. Nature Rev Microbiol. 2009;7:99–109.
Franchi L, Munoz-Planillo R, Nunez G. Sensing and reacting to microbes through the inflammasomes. Nature Immunol. 2012;13:325–32.
Ummanni R, Lehnigk U, Zimmermann U, Woenckhaus C, Walther R, Giebel J. Immunohistochemical expression of caspase-1 and -9, uncleaved caspase-3 and -6, cleaved caspase-3 and -6 as well as Bcl-2 in benign epithelium and cancer of the prostate. Exp Therapeutic Med. 2010;1:47–52.
Winter RN, Kramer A, Borkowski A, Kyprianou N. Loss of caspase-1 and caspase-3 protein expression in human prostate cancer. Cancer Res. 2001;61:1227–322.
Yue Q, Gao G, Zou G, Yu H, Zheng X. Natural products as adjunctive treatment for pancreatic cancer: recent trends and advancements. BioMed Res Int. 2017;2017:8412508.
Yuan R, Hou Y, Sun W, Yu J, Liu X, Niu Y, et al. Natural products to prevent drug resistance in cancer chemotherapy: a review. Ann N Y Acad Sci. 2017;1401:19–27.
Nkengfack AE, Azebaze AG, Waffo AK, Fomum ZT, Meyer M, van Heerden FR. Cytotoxic isoflavones from Erythrina indica. Phytochemistry. 2001;58:1113–20.
Kumar S, Pathania AS, Saxena AK, Vishwakarma RA, Ali A, Bhushan S. The anticancer potential of flavonoids isolated from the stem bark of Erythrina suberosa through induction of apoptosis and inhibition of STAT signaling pathway in human leukemia HL-60 cells. Chem Biol Interact. 2013;205:128–37.
Gao M, Chang Y, Wang X, Ban C, Zhang F. Reduction of COX-2 through modulating miR-124/SPHK1 axis contributes to the antimetastatic effect of alpinumisoflavone in melanoma. Am J Transl Res. 2017;9:986–98.
Wang Y, Yin B, Li D, Wang G, Han X, Sun X. GSDME mediates caspase-3-dependent pyroptosis in gastric cancer. Biochem Biophys Res Commun. 2018;495:1418–25.
Wei Q, Zhu R, Zhu J, Zhao R, Li M. E2-induced activation of the NLRP3 inflammasome triggers pyroptosis and inhibits autophagy in HCC cells. Oncol Res. 2019;27:827–34.
Han M, Gao H, Xie J, Yuan YP, Yuan Q, Gao MQ, et al. Hispidulin induces ER stress-mediated apoptosis in human hepatocellular carcinoma cells in vitro and in vivo by activating AMPK signaling pathway. Acta Pharmacol Sin. 2019;40:666–76.
Yoshii SR, Mizushima N. Monitoring and measuring autophagy. Int J Mol Sci. 2017;2017:18.
Han M, Gao H, Ju P, Gao MQ, Yuan YP, Chen XH, et al. Hispidulin inhibits hepatocellular carcinoma growth and metastasis through AMPK and ERK signaling mediated activation of PPARgamma. Biomed Pharmacotherapy. 2018;103:272–83.
Yu P, Wang HY, Tian M, Li AX, Chen XS, Wang XL, et al. Eukaryotic elongation factor-2 kinase regulates the cross-talk between autophagy and pyroptosis in doxorubicin-treated human melanoma cells in vitro. Acta Pharmacol Sin. 2019;40:123–1244.
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XL designed the research and wrote the paper. YZ, HY, MS, and TH performed the experiments. YL, XY, and XS analyzed the data and collected the information.
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43440_2020_64_MOESM1_ESM.jpg
The effect of AIF on the histological changes of heart, liver, spleen, lung, kidney as detected by H&E staining (JPG 440 kb)
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Zhang, Y., Yang, H., Sun, M. et al. Alpinumisoflavone suppresses hepatocellular carcinoma cell growth and metastasis via NLRP3 inflammasome-mediated pyroptosis. Pharmacol. Rep 72, 1370–1382 (2020). https://doi.org/10.1007/s43440-020-00064-8
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DOI: https://doi.org/10.1007/s43440-020-00064-8