• PANNEER SELVAM CHERMAKANI Cell Biology Laboratory, Department of Biochemistry, University of Madras, Guindy Campus, Chennai 600025, Tamil Nadu, India
  • GANAPASAM SUDHANDIRAN Cell Biology Laboratory, Department of Biochemistry, University of Madras, Guindy Campus, Chennai 600025, Tamil Nadu, India


Objective: The objective of the study is to investigate the effect of escin in hypoxia mimicked NCI-H23 cells through the modulation of matrix metalloproteinases (MMPs) 2 and 9.

Methods: In escin-treated NCI-H23 cells, adhesion, migration, and invasion were detected by the adhesion, wound healing, and Boyden chamber assays, respectively. The activation of proteinases was detected using zymography assay. The expressions of HIF-1α and HIF-2α were evaluated by immunoblot.

Results: In the present study, it was observed that escin suppressed chemically induced hypoxia condition and stimulated adhesion, migration, and invasion of NCI-H23 cells. Gelatin zymography assay showed that escin inhibited CoCl2 induced MMPs-2 and 9 activations in NCI-H23 cells. Furthermore, immunoblot analysis revealed that escin treatment decreased the expression of both HIF-1 and 2α in a dose-dependent manner under CoCl2 induced hypoxia condition.

Conclusion: Taken together, these results indicate that escin inhibits HIFs-α mediated MMPs-2 and 9 expressions, resulting in suppression of lung cancer cell invasion that is induced by chemically induced hypoxia condition. Escin is a potential therapeutic agent for clinical use in preventing the invasion of human malignant lung tumors.

Keywords: Lung cancer, Escin, Hypoxia, Invasion, and MMPs


Download data is not yet available.


1. Mushtaq S, Abbasi BH, Uzair B, Abbasi R. Natural products as reservoirs of novel therapeutic agents. EXCLI J 2018;17:420.
2. Vaskova J, Fejercakova A, Mojzisova G, Vasko L, Patlevic P. Antioxidant potential of aesculus hippocastanum extract and escin against reactive oxygen and nitrogen species. Eur Rev Med Pharmacol Sci 2015;19:879-6.
3. Yuan SY, Cheng CL, Wang SS, Ho HC, Chiu KY, Chen CS, et al. Escin induces apoptosis in human renal cancer cells through G2/M arrest and reactive oxygen species-modulated mitochondrial pathways. Oncol Rep 2017;37:1002-10.
4. Li M, Lu C, Zhang L, Zhang J, Du Y, Duan S, et al. Oral administration of escin inhibits acute inflammation and reduces intestinal mucosal injury in animal models. Evid Based Complement Alternat Med 2015. DOI:10.1155/2015/503617
5. Varinska L, Faber L, Kello M, Petrovova E, Balazova L, Solar P, et al. ?-escin effectively modulates HUVECs proliferation and tube formation. Molecules 2018;23:197.
6. Huang GL, Shen DY, Cai CF, Zhang QY, Ren HY, Chen QX. ?-escin reverses multidrug resistance through inhibition of the GSK3?/?-catenin pathway in cholangiocarcinoma. World J Gastroenterol 2015;21:1148.
7. Kwak H, An H, Alam MB, Choi WS, Lee SY, Lee SH. Inhibition of migration and invasion in melanoma cells by ?-escin via the ERK/NF-?B signaling pathway. Biol Pharm Bull 2018;41:1606-10.
8. Wang Y, Xu X, Zhao P, Tong B, Wei Z, Dai Y. Escin Ia suppresses the metastasis of triple-negative breast cancer by inhibiting epithelial-mesenchymal transition via down-regulating LOXL2 expression. Oncotarget 2016;7:23684.
9. Harikumar KB, Sung B, Pandey MK, Guha S, Krishnan S, Aggarwal BB. Escin, a pentacyclic triterpene, chemosensitizer human tumor cells through inhibition of the nuclear factor-?B signaling pathway. Mol Pharmacol 2010;77:818-27.
10. Nichols L, Saunders R, Knollmann FD. Causes of death of patients with lung cancer. Arch Pathol Lab Med 2012;136:1552-7.
11. Alt?nay S. Is Extracellular matrix a castle against to invasion of cancer cells? In Tumor Metastasis Intech Open; 2016.
12. Klein T, Bischoff R. Physiology and pathophysiology of matrix metalloproteases. Amino Acids 2011;41:271-90.
13. Djuric T, Zivkovic M. Overview of MMP biology and gene associations in human diseases. Role Matrix Met Hum Body Pathol 2017;1:3-3.
14. Mustafa SH, Muhamad M, Ab-rahim SH. Aberrant n-glycosylation regulates invasion of mg-63 cells through extracellular matrix remodeling. Int J Appl Pharm 2019;11:75-9.
15. Roomi MW, Monterrey JC, Kalinovsky T, Rath M, Niedzwiecki A. Patterns of MMP-2 and MMP-9 expression in human cancer cell lines. Oncol Rep 2009;21:1323-33.
16. Gialeli C, Theocharis AD, Karamanos NK. Roles of matrix metalloproteinases in cancer progression and their pharmacological targeting. FEBS J 2011;278:16-27.
17. Miyazaki Y, Hara A, Kato K, Oyama T, Yamada Y, Mori H, et al. The effect of hypoxic microenvironment on matrix metalloproteinase expression in xenografts of human oral squamous cell carcinoma. Int J Oncol 2008;32:145-51.
18. Fu OY, Hou MF, Yang SF, Huang SC, Lee WY. Cobalt chloride-induced hypoxia modulates the invasive potential and matrix metalloproteinases of primary and metastatic breast cancer cells. Anticancer Res 2009;29:3131-8.
19. Shi CY, Fan Y, Liu B, Lou WH. HIF1 contributes to hypoxia-induced pancreatic cancer cells invasion via promoting QSOX1 expression. Cell Physiol Biochem 2013;32:561-8.
20. Li NA, Wang H, Zhang J, Zhao E. Knockdown of hypoxia-inducible factor-2? inhibits cell invasion via the down regulation of MMP-2 expression in breast cancer cells. Oncol Lett 2016;11:3743-8.
21. Merchant N, Nagaraju GP, Rajitha B, Lammata S, Jella KK, Buchwald ZS, et al. Matrix metalloproteinases: their functional role in lung cancer. Carcinogenesis 2017;38:766-80.
22. Lin SS, Lai KC, Hsu SC, Yang JS, Kuo CL, Lin JP, et al. Curcumin inhibits the migration and invasion of human A549 lung cancer cells through the inhibition of matrix metalloproteinase-2 and-9 and vascular endothelial growth factor (VEGF). Cancer Lett 2009;285:127-33.
23. Paneerselvam C, Ganapasam S. ?-Escin alleviates cobalt chloride-induced hypoxia-mediated apoptotic resistance and invasion via ROS-dependent HIF-1?/TGF-?/MMPs in A549 cells. Toxicol Res 2020;9:191-201.
24. Wang J, Huang S. Fisetin inhibits the growth and migration in the A549 human lung cancer cell line via the ERK1/2 pathway. Exp Ther Med 2018;15:2667-73.
25. Lecomte N, Njardarson JT, Nagorny P, Yang G, Downey R, Ouerfelli O, et al. Emergence of potent inhibitors of metastasis in lung cancer via syntheses based on migrastatin. Proc Nat Acad Sci 2011;108:15074-8.
26. Ryabaya O, Prokofieva A, Akasov R, Khochenkov D, Emelyanova M, Burov S, et al. Metformin increases antitumor activity of MEK inhibitor binimetinib in 2D and 3D models of human metastatic melanoma cells. Biomed Pharmacother 2019;109:2548-60.
27. Heo DS, Choi H, Yeom MY, Song BJ, Oh SJ. Serum levels of matrix metalloproteinase-9 predict lymph node metastasis in breast cancer patients. Oncol Rep 2014;31:1567-72.
28. Janiszewska M, Primi MC, Izard T. Cell adhesion in cancer: beyond the migration of single cells. J Biol Chem 2020;295:2495-505.
29. Pachmayr E, Treese C, Stein U. Underlying mechanisms for distant metastasis-molecular biology. Visc Med 2017;33:11-20.
30. Shay G, Lynch CC, Fingleton B. Moving targets: emerging roles for MMPs in cancer progression and metastasis. Matrix Biol 2015;44:200-6.
31. Finger EC, Giaccia AJ. Hypoxia, inflammation, and tumor microenvironment in metastatic disease. Cancer Metastasis Rev 2010;29:285-93.
32. Semenza GL. The hypoxic tumor microenvironment: a driving force for breast cancer progression. Biochim Biophys Acta 2016;1863:382-91.
33. Martin TA, Ye L, Sanders AJ, Lane J, Jiang WG. Cancer invasion and metastasis: molecular and cellular perspective. In: Madame Curie Bioscience Database [Internet] Landes Bioscience; 2013.
34. Keith B, Johnson RS, Simon MC. HIF1? and HIF2?: sibling rivalry in hypoxic tumour growth and progression. Nat Rev Cancer 2012;12:9-22.
35. Wigerup C, Pahlman S, Bexell D. Therapeutic targeting of hypoxia and hypoxia-inducible factors in cancer. Pharmacol Ther 2016;164:152-69.
36. Lu X, Kang Y. Hypoxia and hypoxia-inducible factors: master regulators of metastasis. Clin Cancer Res 2010;16:5928-35.
37. Biswas S, Troy H, Leek R, Chung YL, Li JL, Raval RR, et al. Effects of HIF-1? and HIF2? on growth and metabolism of clear-cell renal cell carcinoma 786-0 xenografts. J Oncol 2010. DOI:10.1155/2010/757908
38. Zhao J, Du F, Shen G, Zheng F, Xu B. The role of hypoxia-inducible factor-2 in digestive system cancers. Cell Death Dis 2015;6:e1600.
39. Abbas S, Malla S. Cytotoxicity and expression studies of angiogenesis-promoting genes in cancer cell lines under the treatment of cancer candidate drugs. Asian J Pharm Clin Res 2019;12:130-4.
40. Wan R, Mo Y, Chien S, Li Y, Li Y, Tollerud DJ, et al. The role of hypoxia-inducible factor-1? in the increased MMP-2 and MMP-9 production by human monocytes exposed to nickel nanoparticles. Nanotoxicology 2011;5:568-82.
41. Cai X, Zhu H, Li Y. PKC?, MMP 2 and MMP 9 expression in lung adenocarcinoma and association with a metastatic phenotype. Mol Med Rep 2017;16:8301-6.
42. Lopez Sanchez LM, Jimenez C, Valverde A, Hernandez V, Penarando J, Martinez A, et al. CoCl2, a mimic of hypoxia, induces the formation of polyploid giant cells with stem characteristics in colon cancer. PloS One 2014;9:e99143.
43. Bauer N, Liu L, Aleksandrowicz E, Herr I. Establishment of hypoxia induction in an in vivo animal replacement model for experimental evaluation of pancreatic cancer. Oncol Rep 2014;32:153-8.
44. Bedessem B, Marie Paule M, Hamel M, Giroud F, Stephanou A. Effects of the hypoxia-mimetic agents DFO and CoCl2 on HeLa-fucci cells. J Cell Biol Cell Metab 2015;2:1.
45. Yuan Y, Hilliard G, Ferguson T, Millhorn DE. Cobalt inhibits the interaction between hypoxia-inducible factor-? and von hippel-lindau protein by direct binding to hypoxia-inducible factor-?. J Biol Chem 2003;278:15911-6.
46. Taheem DK, Foyt DA, Loaiza S, Ferreira SA, Ilic D, Auner HW, et al. Differential regulation of human bone marrow mesenchymal stromal cell chondrogenesis by hypoxia-inducible factor?1? hydroxylase inhibitors. Stem Cells 2018;36:1380-92.
47. Fang Y, Zhang H, Zhong Y, Ding X. Prolyl hydroxylase 2 (PHD2) inhibition protects human renal epithelial cells and mice kidney from hypoxia injury. Oncotarget 2016;7:54317.
48. Shyu KG, Hsu FL, Wang MJ, Wang BW, Lin S. Hypoxia-inducible factor 1? regulates lung adenocarcinoma cell invasion. Exp Cell Res 2007;313:1181-91.
49. Fujiwara S, Nakagawa KO, Harada H, Nagato S, Furukawa K, Teraoka M, et al. Silencing hypoxia-inducible factor-1? inhibits cell migration and invasion under hypoxic environment in malignant gliomas. Int J Oncol 2007;30:793-802.
50. Chu CY, Jin YT, Zhang W, Yu J, Yang HP, Wang HY, et al. CA IX is upregulated in CoCl2-induced hypoxia and associated with cell invasive potential and a poor prognosis of breast cancer. Int J Oncol 2015;48:271-80.
51. Lu N, Zhou H, Lin YH, Chen ZQ, Pan Y, Li XJ. Oxidative stress mediates CoCl2 induced prostate tumour cell adhesion: Role of protein kinase c and p38 mitogen?activated protein kinase. Basic Clin Pharmacol Toxicol 2007;101:41-6.
125 Views | 118 Downloads
How to Cite
CHERMAKANI, P. S., and G. SUDHANDIRAN. “ESCIN MITIGATES HYPOXIA MIMICKING NCI-H23 CELLS THROUGH MODULATION OF MMPs, HIF-1α AND HIF-2α”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 12, no. 12, Dec. 2020, pp. 26-30, doi:10.22159/ijpps.2020v12i12.39788.
Original Article(s)