Abstract
The cytostatic drug from fruits and other plant derived products have acted as a chemotherapeutic agent used in treatment of a wide variety of cancers. Lupeol, a dietary triterpene, present in many fruits and medicinal plants, has been shown to possess many pharmacological properties including anti-cancer effect in both in vitro and in vivo assay systems. However, the cancer proliferative and invasive inhibitory effects and molecular mechanisms on gallbladder carcinoma GBC-SD cells have not been studied. In the present study, GBC-SD cells were treated by lupeol and subjected to methyl thiazolyl tetrazolium analysis, Hoechst 33342 staining, annexin V/propidium iodide double-staining, transwell chamber assay and Western blot analysis. In addition, GBC-SD xenograft tumors were established in male nude BALB/c mice, and lupeol was intravenously administered to evaluate the anti-cancer capacity in vivo. Our results showed that lupeol inhibited the proliferation, migration, invasion and induced apoptosis of GBC-SD cells in a dose-dependent manner in vitro. Furthermore, the expression of p-EGFR, p-AKT and MMP-9 levels were significantly down-regulated. These protein interactions may play a pivotal role in the regulation of apoptosis and invasion. More importantly, our in vivo studies showed that administration of lupeol decreased tumor growth in a dose-dependent manner. Immunohistochemistry analysis demonstrated the down-regulation of p-EGFR and MMP-9 in tumor tissues following lupeol treatment, consistent with the in vitro results. Taken together, our findings indicated that lupeol can induce apoptotic cell death and inhibit the migration as well as invasion of GBC-SD cells. The mechanism may be associated with the suppression of EGFR/MMP-9 signaling. These results might offer a therapeutic potential advantage for human gallbladder carcinoma chemoprevention or chemotherapy.
Similar content being viewed by others
Change history
03 October 2022
This article has been retracted. Please see the Retraction Notice for more detail: https://doi.org/10.1007/s10616-022-00550-2
References
Ardiansyah Yamaguchi E, Shirakawa H, Hata K, Hiwatashi K, Ohinata K, Goto T, Komai M (2012) Lupeol supplementation improves blood pressure and lipid metabolism parameters in stroke-prone spontaneously hypertensive rats. Biosci Biotechnol Biochem 76:183–185
Ayyappan S, Prabhakar D, Sharma N (2013) Epidermal growth factor receptor (EGFR)-targeted therapies in esophagogastric cancer. Anticancer Res 33:4139–4155
Chaturvedi PK, Bhui K, Shukla Y (2008) Lupeol: connotations for chemoprevention. Cancer Lett 263:1–13. doi:10.1016/j.canlet.2008.01.047
Chi A, Remick S, Tse W (2013) EGFR inhibition in non-small cell lung cancer: current evidence and future directions. Biomark Res 1:2. doi:10.1186/2050-7771-1-2
Hlobilkova A, Ehrmann J, Knizetova P, Krejci V, Kalita O, Kolar Z (2009) Analysis of VEGF, Flt-1, Flk-1, nestin and MMP-9 in relation to astrocytoma pathogenesis and progression. Neoplasma 56:284–290
Imam S, Azhar I, Hasan MM, Ali MS, Ahmed SW (2007) Two triterpenes lupanone and lupeol isolated and identified from Tamarindus indica linn. Pak J Pharm Sci 20:125–127
Jia JG, Li SG, Gong W, Ding J, Fang CF, Quan ZW (2011) mda-7/IL-24 induces apoptosis in human GBC-SD gallbladder carcinoma cells via mitochondrial apoptotic pathway. Oncol Rep 25:195–201. doi:10.3892/or_00001061
Kelly PN, Strasser A (2011) The role of Bcl-2 and its pro-survival relatives in tumourigenesis and cancer therapy. Cell Death Differ 18:1414–1424. doi:10.1038/cdd.2011.17
Khan N, Afaq F, Mukhtar H (2008) Cancer chemoprevention through dietary antioxidants: progress and promise. Antioxid Redox Signal 10:475–510. doi:10.1089/ars.2007.1740
Liang CZ, Zhang JK, Shi Z, Liu B, Shen CQ, Tao HM (2012) Matrine induces caspase-dependent apoptosis in human osteosarcoma cells in vitro and in vivo through the upregulation of Bax and Fas/FasL and downregulation of Bcl-2. Cancer Chemother Pharmacol 69:317–331. doi:10.1007/s00280-011-1699-4
Liu F, He Y, Liang Y, Wen L, Zhu Y, Wu Y, Zhao L, Li Y, Mao X, Liu H (2013a) PI3-kinase inhibition synergistically promoted the anti-tumor effect of lupeol in hepatocellular carcinoma. Cancer Cell Int 13:108. doi:10.1186/1475-2867-13-108
Liu TY, Tan ZJ, Jiang L, Gu JF, Wu XS, Cao Y, Li ML, Wu KJ, Liu YB (2013b) Curcumin induces apoptosis in gallbladder carcinoma cell line GBC-SD cells. Cancer Cell Int 13:64. doi:10.1186/1475-2867-13-64
Marshall J (2006) Clinical implications of the mechanism of epidermal growth factor receptor inhibitors. Cancer 107:1207–1218. doi:10.1002/cncr.22133
Martini M, Ciraolo E, Gulluni F, Hirsch E (2013) Targeting PI3K in cancer: any good news? Front Oncol 3:108. doi:10.3389/fonc.2013.00108
Mitsudomi T, Yatabe Y (2010) Epidermal growth factor receptor in relation to tumor development: EGFR gene and cancer. FEBS J 277:301–308. doi:10.1111/j.1742-4658.2009.07448.x
Mlcochova J, Faltejskova P, Nemecek R, Svoboda M, Slaby O (2013) MicroRNAs targeting EGFR signalling pathway in colorectal cancer. J Cancer Res Clin Oncol 139:1615–1624. doi:10.1007/s00432-013-1470-9
Murtaza I, Saleem M, Adhami VM, Hafeez BB, Mukhtar H (2009) Suppression of cFLIP by lupeol, a dietary triterpene, is sufficient to overcome resistance to TRAIL-mediated apoptosis in chemoresistant human pancreatic cancer cells. Cancer Res 69:1156–1165. doi:10.1158/0008-5472.can-08-2917
Park JH, Jeong YJ, Park KK, Cho HJ, Chung IK, Min KS, Kim M, Lee KG, Yeo JH, Park KK, Chang YC (2010) Melittin suppresses PMA-induced tumor cell invasion by inhibiting NF-kappaB and AP-1-dependent MMP-9 expression. Mol Cells 29:209–215. doi:10.1007/s10059-010-0028-9
Prasad S, Madan E, Nigam N, Roy P, George J, Shukla Y (2009) Induction of apoptosis by lupeol in human epidermoid carcinoma A431 cells through regulation of mitochondrial, Akt/PKB and NF-kappaB signaling pathways. Cancer Biol Ther 8:1632–1639
Qiu Q, Yang M, Tsang BK, Gruslin A (2004) EGF-induced trophoblast secretion of MMP-9 and TIMP-1 involves activation of both PI3K and MAPK signalling pathways. Reproduction 128:355–363. doi:10.1530/rep.1.00234
Reid KM, Ramos-De la Medina A, Donohue JH (2007) Diagnosis and surgical management of gallbladder cancer: a review. J Gastrointest Surg 11:671–681. doi:10.1007/s11605-006-0075-x
Saleem M (2009) Lupeol, a novel anti-inflammatory and anti-cancer dietary triterpene. Cancer Lett 285:109–115. doi:10.1016/j.canlet.2009.04.033
Saleem M, Afaq F, Adhami VM, Mukhtar H (2004) Lupeol modulates NF-kappaB and PI3K/Akt pathways and inhibits skin cancer in CD-1 mice. Oncogene 23:5203–5214. doi:10.1038/sj.onc.1207641
Saleem M, Kaur S, Kweon MH, Adhami VM, Afaq F, Mukhtar H (2005) Lupeol, a fruit and vegetable based triterpene, induces apoptotic death of human pancreatic adenocarcinoma cells via inhibition of Ras signaling pathway. Carcinogenesis 26:1956–1964. doi:10.1093/carcin/bgi157
Saleem M, Murtaza I, Tarapore RS, Suh Y, Adhami VM, Johnson JJ, Siddiqui IA, Khan N, Asim M, Hafeez BB, Shekhani MT, Li B, Mukhtar H (2009) Lupeol inhibits proliferation of human prostate cancer cells by targeting beta-catenin signaling. Carcinogenesis 30:808–817. doi:10.1093/carcin/bgp044
Taner CB, Nagorney DM, Donohue JH (2004) Surgical treatment of gallbladder cancer. J Gastrointest Surg 8:83–89 (discussion 89)
Tarapore RS, Siddiqui IA, Saleem M, Adhami VM, Spiegelman VS, Mukhtar H (2010) Specific targeting of Wnt/beta-catenin signaling in human melanoma cells by a dietary triterpene lupeol. Carcinogenesis 31:1844–1853. doi:10.1093/carcin/bgq169
Tian YC, Chen YC, Chang CT, Hung CC, Wu MS, Phillips A, Yang CW (2007) Epidermal growth factor and transforming growth factor-beta1 enhance HK-2 cell migration through a synergistic increase of matrix metalloproteinase and sustained activation of ERK signaling pathway. Exp Cell Res 313:2367–2377. doi:10.1016/j.yexcr.2007.03.022
Tomas A, Futter CE, Eden ER (2014) EGF receptor trafficking: consequences for signaling and cancer. Trends Cell Biol 24:26–34. doi:10.1016/j.tcb.2013.11.002
Wachsberger PR, Lawrence YR, Liu Y, Rice B, Feo N, Leiby B, Dicker AP (2014) Hsp90 inhibition enhances PI-3 kinase inhibition and radiosensitivity in glioblastoma. J Cancer Res Clin Oncol 140:573–582. doi:10.1007/s00432-014-1594-6
Wang S, Yu S, Shi W, Ge L, Yu X, Fan J, Zhang J (2011) Curcumin inhibits the migration and invasion of mouse hepatoma Hca-F cells through down-regulating caveolin-1 expression and epidermal growth factor receptor signaling. IUBMB Life 63:775–782. doi:10.1002/iub.507
Washiro M, Ohtsuka M, Kimura F, Shimizu H, Yoshidome H, Sugimoto T, Seki N, Miyazaki M (2008) Upregulation of topoisomerase IIalpha expression in advanced gallbladder carcinoma: a potential chemotherapeutic target. J Cancer Res Clin Oncol 134:793–801. doi:10.1007/s00432-007-0348-0
Yang L, Yang L, Tian W, Li J, Liu J, Zhu M, Zhang Y, Yang Y, Liu F, Zhang Q, Liu Q, Shen Y, Qi Z (2014) Resveratrol plays dual roles in pancreatic cancer cells. J Cancer Res Clin Oncol. doi:10.1007/s00432-014-1624-4
Yilmaz M, Christofori G, Lehembre F (2007) Distinct mechanisms of tumor invasion and metastasis. Trends Mol Med 13:535–541. doi:10.1016/j.molmed.2007.10.004
You YJ, Nam NH, Kim Y, Bae KH, Ahn BZ (2003) Antiangiogenic activity of lupeol from Bombax ceiba. Phytother Res 17:341–344. doi:10.1002/ptr.1140
Yu PF, Liu Q, Liu K, Yagasaki K, Wu E, Zhang GY (2009) Matrine suppresses breast cancer cell proliferation and invasion via VEGF-Akt-NF-kappaB signaling. Cytotechnology 59:219–229. doi:10.1007/s10616-009-9225-9
Zhang Y, Zhang H, Yu P, Liu Q, Liu K, Duan H, Luan G, Yagasaki K, Zhang G (2009) Effects of matrine against the growth of human lung cancer and hepatoma cells as well as lung cancer cell migration. Cytotechnology 59:191–200. doi:10.1007/s10616-009-9211-2
Conflict of interest
The authors declare no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Additional information
Yan Liu and Tingting Bi contributed equally to this work.
About this article
Cite this article
Liu, Y., Bi, T., Shen, G. et al. RETRACTED ARTICLE: Lupeol induces apoptosis and inhibits invasion in gallbladder carcinoma GBC-SD cells by suppression of EGFR/MMP-9 signaling pathway. Cytotechnology 68, 123–133 (2016). https://doi.org/10.1007/s10616-014-9763-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10616-014-9763-7