Inhibitory Effect of Damulin B from Gynostemma pentaphyllum on Human Lung Cancer Cells

Abstract

Damulin B, a dammarane-type saponin from steamed Gynostemma pentaphyllum, exhibits the strongest activity against human lung carcinoma A549 cells among the isolated active saponins. In this study, the structure-activity relationship of a series of saponin compounds was discussed. The inhibitory effect of damulin B on human lung cancer A549 and H1299 cells was investigated from apoptosis, cell cycle, and migration aspects. In vitro, human lung cancer cells were more susceptible to damulin B treatment than human normal fibroblasts. Damulin B exhibited a strong cytotoxic effect, as evidenced by the increase of apoptosis rate, reduction of mitochondrial membrane potential (MMP), generation of reactive oxygen species, and G0/G1 phase arrest. Furthermore, damulin B activated the following: both intrinsic and extrinsic apoptosis pathways along with early G1 phase arrest via the upregulation of the Bax, Bid, tBid, cleaved caspase-8, and p53 expression levels; downregulation of the procaspase-8/-9, CDK4, CDK6, and cyclin D1 expression levels; and more release of cytochrome c in the cytoplasm. In addition, antimigratory activities and suppressive effects on metastasis-related factors, such as MMP-2 and MMP-9, accompanied by the upregulation of IL-24 were revealed. Altogether, the results proved that damulin B could inhibit human lung cancer cells by inducing apoptosis, blocking the cell cycle at early G0/G1 phase and suppressing the migration. Hence, damulin B has potential therapeutic efficacy against lung cancer.

Key words

Gynostemma pentaphyllum - Curcubitaceae - saponin - damulin B - cytotoxicity - lung cancer

^* Authorship note: Shao-Fang Xing and Lin-Hua Liu contributed equally to this work.

• References

• 1 Siegel RL, Miller KD, Jemal A. Cancer statistics, 2015. CA Cancer J Clin 2015; 65: 5-29
• 2 Guo Y, Zeng H, Zheng R, Li S, Barnett AG, Zhang S, Zou X, Huxley R, Chen W, Williams G. The association between lung cancer incidence and ambient air pollution in China: a spatiotemporal analysis. Environ Res 2015; 144: 60-65
• 3 Shibuya K, Mathers CD, Boschi-Pinto C, Lopez AD, Murray CJ. Global and regional estimates of cancer mortality and incidence by site: II. Results for the global burden of disease 2000. BMC Cancer 2002; 2: 37
• 4 Potiron VA, Roche J, Drabkin HA. Semaphorins and their receptors in lung cancer. Cancer Lett 2009; 273: 1-14
• 5 Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol 2007; 35: 495-516
• 6 Hengartner MO. The biochemistry of apoptosis. Nature 2000; 407: 770-776
• 7 van Gurp M, Festjens N, van Loo G, Saelens X, Vandenabeele P. Mitochondrial intermembrane proteins in cell death. Biochem Biophys Res Commun 2003; 304: 487-497
• 8 Cory S, Adams JM. The Bcl2 family: regulators of the cellular life-or-death switch. Nat Rev Cancer 2002; 2: 647-656
• 9 Everett H, McFadden G. Viruses and apoptosis: meddling with mitochondria. Virology 2001; 288: 1-7
• 10 Malumbres M, Barbacid M. To cycle or not to cycle: a critical decision in cancer. Nat Rev Cancer 2001; 1: 222-231
• 11 Lam M, Carmichael AR, Griffiths HR. An aqueous extract of Fagonia cretica induces DNA damage, cell cycle arrest and apoptosis in breast cancer cells via FOXO3a and p53 expression. PLoS One 2012; 7: e40152
• 12 Fidler IJ. Critical determinants of metastasis. Semin Cancer Biol 2002; 12: 89-96
• 13 Lebedeva IV, Su ZZ, Sarkar D, Kitada S, Dent P, Waxman S, Reed JC, Fisher PB. Melanoma differentiation associated gene-7, mda-7/interleukin-24, induces apoptosis in prostate cancer cells by promoting mitochondrial dysfunction and inducing reactive oxygen species. Cancer Res 2003; 63: 8138-8144
• 14 Gupta P, Su ZZ, Lebedeva IV, Sarkar D, Sauane M, Emdad L, Bachelor MA, Grant S, Curiel DT, Dent P, Fisher PB. MDA-7/IL-24: multifunctional cancer-specific apoptosis-inducing cytokine. Pharmacol Ther 2006; 111: 596-628
• 15 Ramesh R, Ito I, Gopalan B, Saito Y, Mhashilkar AM, Chada S. Ectopic production of MDA-7/IL-24 inhibits invasion and migration of human lung cancer cells. Mol Ther 2004; 9: 510-518
• 16 Huang TH, Razmovski-Naumovski V, Salam NK, Duke RK, Tran VH, Duke CC, Roufogalis BD. A novel LXR-alpha activator identified from the natural product Gynostemma pentaphyllum . Biochem Pharmacol 2005; 70: 1298-1308
• 17 Liang QC, Zhong M. Zhuang Medicine. Guangxi: Guangxi Nationalities Publishing House; 2005: 94-96
• 18 Zhao GP, Dai S, Chen RS. Dictionary of traditional Chinese Medicine. 2nd ed.. Shanghai: Shanghai Scientific & Technical Publishers; 2006: 2460-2462
• 19 Wang JR, Zhao JB. The effect of preventing recurrence of cancer metastasis on jiaogulan soup in clinical study. Zhejiang Zhong Yi Za Zhii 1993; 28: 529-530
• 20 Kao TH, Huang SC, Inbaraj BS, Chen BH. Determination of flavonoids and saponins in Gynostemma pentaphyllum (Thunb.) Makino by liquid chromatography-mass spectrometry. Anal Chim Acta 2008; 626: 200-211
• 21 Piao XL, Wu Q, Yang J, Park SY, Chen DJ, Liu HM. Dammarane-type saponins from heat-processed Gynostemma pentaphyllum show fortified activity against A549 cells. Arch Pharm Res 2013; 36: 874-879
• 22 Chen WQ, Zeng HM, Jemal A, Yu XQ, He J. Cancer statistics in China, 2015. CA Cancer J Clin 2016; 2: 115-132
• 23 Piao XL, Xing SF, Lou CX, Chen DJ. Novel dammarane saponins from Gynostemma pentaphyllum and their cytotoxic activities against HepG2 cells. Bioorg Med Chem Lett 2014; 24: 4831-4833
• 24 Xing SF, Chen DJ, Liu HM, Piao XL. Isolation and identification of ginsenoside Rg3 from heat-processed Gocaekmbaw of Zhuang-medicine. Chin J Exp Tradit Med Form 2014; 20: 120-124
• 25 Xing SF, Jang M, Wang YR, Piao XL. A new dammarane-type saponin from Gynostemma pentaphyllum induces apoptosis in A549 human lung carcinoma cells. Bioorg Med Chem Lett 2016; 26: 1754-1759
• 26 Gottlieb E, Armour SM, Harris MH, Thompson CB. Mitochondrial membrane potential regulates matrix configuration and cytochrome c release during apoptosis. Cell Death Differ 2003; 10: 709-717
• 27 Maillet A, Yadav S, Loo YL, Sachaphibulkij K, Pervaiz S. A novel osmium-based compound targets the mitochondria and triggers ROS-dependent apoptosis in colon carcinoma. Cell Death Dis 2013; 4: e653
• 28 Kim KM, Heo DR, Lee J, Park JS, Baek MG, Yi JM, Kim H, Bang OS. 5,3′-Dihydroxy-6,7,4′-trimethoxyflavanone exerts its anticancer and antiangiogenesis effects through regulation of the Akt/mTOR signaling pathway in human lung cancer cells. Chem Biol Interact 2015; 225: 32-39
• 29 Green DR, Reed JC. Mitochondria and apoptosis. Science 1998; 281: 1309-1312
• 30 Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng TI, Jones DP, Wang X. Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science 1997; 275: 1129-1132
• 31 Hossini AM, Eberle J. Apoptosis induction by Bcl-2 proteins independent of the BH3 domain. Biochem Pharmacol 2008; 76: 1612-1619
• 32 Li H, Zhu H, Xu CJ, Yuan J. Cleavage of BID by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis. Cell 1998; 94: 491-501
• 33 Gross A, Yin XM, Wang K, Wei MC, Jockel J, Milliman C, Erdjument-Bromage H, Tempst P, Korsmeyer SJ. Caspase cleaved BID targets mitochondria and is required for cytochrome c release, while BCL-XL prevents this release but not tumor necrosis factor-R1/Fas death. J Biol Chem 1999; 274: 1156-1163
• 34 Eskes R, Desagher S, Antonsson B, Martinou JC. Bid induces the oligomerization and insertion of Bax into the outer mitochondrial membrane. Mol Cell Biol 2000; 20: 929-935
• 35 Wei MC, Lindsten T, Mootha VK, Weiler S, Gross A, Ashiya M, Thompson CB, Korsmeyer SJ. tBID, a membrane-targeted death ligand, oligomerizes BAK to release cytochrome c. Genes Dev 2000; 14: 2060-2071
• 36 Wei C, Xiao Q, Kuang X, Zhang T, Yang Z, Wang L. Fucoidan inhibits proliferation of the SKM-1 acute myeloid leukaemia cell line via the activation of apoptotic pathways and production of reactive oxygen species. Mol Med Rep 2015; 12: 6649-6655
• 37 Seo KH, Ryu HW, Park MJ, Park KH, Kim JH, Lee MJ, Kang HJ, Kim SL, Lee JH, Seo WD. Mangosenone F, A furanoxanthone from Garciana mangostana, induces reactive oxygen species-mediated apoptosis in lung cancer cells and decreases xenograft tumor growth. Phytother Res 2015; 29: 1753-1760
• 38 Liu G, Kuang S, Wu S, Jin W, Sun C. A novel polysaccharide from Sargassum integerrimum induces apoptosis in A549 cells and prevents angiogensis in vitro and in vivo . Sci Rep 2016; 6: 26722
• 39 Schwartz GK, Shah MA. Targeting the cell cycle: a new approach to cancer therapy. J Clin Oncol 2005; 23: 9408-9421
• 40 Yang JB, Khan M, He YY, Yao M, Li YM, Gao HW, Ma TH. Tubeimoside-1 induces oxidative stress-mediated apoptosis and G0/G1 phase arrest in human prostate carcinoma cells in vitro . Acta Pharmacol Sin 2016; 37: 950-962
• 41 Khan M, Yu B, Rasul A, Al Shawi A, Yi F, Yang H, Ma T. Jaceosidin induces apoptosis in U87 glioblastoma cells through G2/M phase arrest. Evid Based Complement Alternat Med 2012; 2012: 703034
• 42 Matrisian LM. The matrix-degrading metalloproteinases. Bioessays 1992; 14: 455-463
• 43 Shapiro SD. Matrix metalloproteinase degradation of extracellular matrix: biological consequences. Curr Opin Cell Biol 1998; 10: 602-608