Gypenosides inhibits migration and invasion of human oral cancer SAS cells through the inhibition of matrix metalloproteinase-2 -9 and urokinase-plasminogen by ERK1/2 and NF-kappa B signaling pathways

Abstract

Gypenosides (Gyp), found in Gynostemma pentaphyllum Makino, has been used as a folk medicine in the Chinese population for centuries and is known to have diverse pharmacologic effects, including anti-proliferative and anti-cancer actions. However, the effects of Gyp on prevention from invasion and migration of oral cancer cells are still unsatisfactory. The purpose of this study was to investigate effects of Gyp treatment on migration and invasion of SAS human oral cancer cells. SAS cells were cultured in the presence of 90 and 180 μg/mL Gyp for 24 and 48 hours. Gyp induced cytotoxic effects and inhibited SAS cells migration and invasion in dose- and time-dependent response. Wound-healing assay and boyden chamber assay were carried out to investigate Gyp-inhibited migration and invasion of SAS cells. Gyp decreased the abundance of several proteins, including nuclear factor-kappa B (NF-κB), cyclooxygenase-2 (COX-2), extracellular signal-regulated kinase 1/2 (ERK1/ 2), matrix metalloproteinase-9, -2 (MMP-9, -2), sevenless homolog (SOS), Ras, urokinase-type plasminogen activator (uPA), focal adhesion kinase (FAK) and RAC-alpha serine/threonine-protein kinase (Akt), in a time-dependent manner. In addition, Gyp decreased mRNA levels of MMP-2, MMP-7, MMP-9 but did not affect FAK and Rho A mRNA levels in SAS cells. These results provide evidences for the role of Gyp as a potent anti-metastatic agent, which can markedly inhibit the metastatic and invasive capacity of oral cancer cells. The inhibition of NF-κB and MMP-2, -7 and -9 signaling may be one of the mechanisms that is present in Gyp-inhibited cancer cell invasion and migration.

Keywords

gypenosides cell migration cell invasion matrix metalloproteinase-2 -9 urokinase-plasminogen SAS cells

References

Chen JC , Lu KW , Tsai ML , et al. Gypenosides induced G0/G1 arrest via CHk2 and apoptosis through endoplasmic reticulum stress and mitochondria-dependent pathways in human tongue cancer SCC-4 cells. Oral Oncol 2009 ; 45: 273-283.

Pintos J. , Black MJ , Sadeghi N. , et al. Human papillomavirus infection and oral cancer: a case-control study in Montreal, Canada. Oral Oncol 2008 ; 44: 242-250.

Shingyoji M. , Ando S. , Nishimura H. , et al. VEGF in patients with non-small cell lung cancer during combination chemotherapy of carboplatin and paclitaxel . Anticancer Res 2009; 29: 2635-2639.

Cui J. , Eneroth P. , Bruhn JG Gynostemma pentaphyllum: identification of major sapogenins and differentiation from Panax species. Eur J Pharm Sci 1999; 8: 187-191.

Shang L. , Liu J. , Zhu Q. , et al. Gypenosides protect primary cultures of rat cortical cells against oxidative neurotoxicity. Brain Res 2006; 1102: 163-174.

Megalli S. , Davies NM , Roufogalis BD Antihyperlipidemic and hypoglycemic effects of Gynostemma pentaphyllum in the Zucker fatty rat. J Pharm Pharm Sci 2006 ; 9: 281-291.

Circosta C. , De Pasquale R. , Occhiuto F. Cardiovascular effects of the aqueous extract of Gynostemma pentaphyllum Makino . Phytomedicine 2005; 12: 638-643.

Aktan F. , Henness S. , Roufogalis BD , et al. Gypenosides derived from Gynostemma pentaphyllum suppress NO synthesis in murine macrophages by inhibiting iNOS enzymatic activity and attenuating NF-kappa B-mediated iNOS protein expression. Nitric Oxide 2003; 8: 235-242.

Wang QF , Chen JC , Hsieh SJ , et al. Regulation of Bcl-2 family molecules and activation of caspase cascade involved in gypenosides-induced apoptosis in human hepatoma cells . Cancer Lett 2002; 183: 169-178.

10.

Hu L. , Chen Z. , Xie Y. New triterpenoid saponins from Gynostemma pentaphyllum. J Nat Prod 1996; 59: 1143-1145.

11.

Wang C. , Wang X. , Li Y. , et al. A preliminary observation of preventive and blocking effect of Gynostemma pentaphyllum (Thunb) Makino on esophageal cancer in rats. Hua Xi Yi Ke Da Xue Xue Bao 1995; 26: 430-432.

12.

Zhou Z. , Wang Y. , Zhou Y. The effect of Gynostemma pentaphyllum mak (GP) on carcinogenesis of the golden hamster cheek pouch induced by DMBA. Zhonghua Kou Qiang Yi Xue Za Zhi 1996; 31: 267-270.

13.

Chiu TH , Chen JC , Chung JG N-acetyltransferase is involved in gypenosides-induced N-acetylation of 2-aminofluorene and DNA adduct formation in human cervix epidermoid carcinoma cells (Ca Ski) . In Vivo 2003; 17: 281-288.

14.

Chen JC , Lu KW , Lee JH , et al. Gypenosides induced apoptosis in human colon cancer cells through the mitochondria-dependent pathways and activation of caspase-3. Anticancer Res 2006; 26: 4313-4326.

15.

Lai WW , Yang JS , Lai KC , et al. Rhein induced apoptosis through the endoplasmic reticulum stress, caspase- and mitochondria-dependent pathways in SCC-4 human tongue squamous cancer cells. In Vivo 2009; 23: 309-316.

16.

Lin SY , Lai WW , Ho CC , et al. Emodin induces apoptosis of human tongue squamous cancer SCC-4 cells through reactive oxygen species and mitochondria-dependent pathways . Anticancer Res 2009; 29: 327-335.

17.

Ho YT , Yang JS , Li TC , et al. Berberine suppresses in vitro migration and invasion of human SCC-4 tongue squamous cancer cells through the inhibitions of FAK, IKK, NF-kappa-B, u-PA and MMP-2 and -9. Cancer Lett 2009; 279: 155-162.

18.

Fishman DA , Liu Y. , Ellerbroek SM , et al. Lysophosphatidic acid promotes matrix metalloproteinase (MMP) activation and MMP-dependent invasion in ovarian cancer cells. Cancer Res 2001; 61: 3194-3199.

19.

Hsu SC , Kuo CL , Lin JP , et al. Crude extracts of Euchresta formosana radix inhibit invasion and migration of human hepatocellular carcinoma cells. Anticancer Res 2007; 27: 2377-2384.

20.

Lin SS , Lai KC , Hsu SC , 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: 127-133.

21.

Zhang X. , Liu Y. , Gilcrease MZ , et al. A lymph node metastatic mouse model reveals alterations of metastasis-related gene expression in metastatic human oral carcinoma sublines selected from a poorly metastatic parental cell line. Cancer 2002; 95: 1663-1672.

22.

Woodhouse EC , Chuaqui RF , Liotta LA General mechanisms of metastasis. Cancer 1997; 80: 1529-1537.

23.

Thomas GT , Lewis MP , Speight PM Matrix metalloproteinases and oral cancer. Oral Oncol 1999; 35: 227-233.

24.

Stacker SA , Achen MG , Jussila L. , et al. Lymphangiogenesis and cancer metastasis. Nat Rev Cancer 2002; 2: 573-583.

25.

Tryggvason K. , Hoyhtya M. , Salo T. Proteolytic degradation of extracellular matrix in tumor invasion. Biochim Biophys Acta 1987 ; 907: 191-217.

26.

Pepper MS Role of the matrix metalloproteinase and plasminogen activator-plasmin systems in angiogenesis. Arterioscler Thromb Vasc Biol 2001; 21: 1104-1117.

27.

Yoshizaki T. , Maruyama Y. , Sato H. , et al. Expression of tissue inhibitor of matrix metalloproteinase-2 correlates with activation of matrix metalloproteinase-2 and predicts poor prognosis in tongue squamous cell carcinoma. Int J Cancer 2001; 95: 44-50.

28.

Jeon YK , Lee BY , Kim JE , et al. Molecular characterization of Epstein-Barr virus and oncoprotein expression in nasopharyngeal carcinoma in Korea. Head Neck 2004; 26: 573-583.

29.

Pacheco MM , Kowalski LP , Nishimoto IN , et al. Differential expression of c-jun and c-fos mRNAs in squamous cell carcinoma of the head and neck: associations with uPA, gelatinase B, and matrilysin mRNAs. Head Neck 2002; 24: 24-32.

30.

Andreasen PA , Kjoller L. , Christensen L. , et al. The urokinase-type plasminogen activator system in cancer metastasis: a review. Int J Cancer 1997; 72: 1-22.

31.

Kwaan HC The plasminogen-plasmin system in malignancy. Cancer Metastasis Rev 1992; 11: 291- 311 .

32.

Blasi F. The urokinase receptor. A cell surface, regulated chemokine. APMIS 1999; 107: 96-101.

33.

Duffy MJ , Maguire TM , McDermott EW , et al. Urokinase plasminogen activator: a prognostic marker in multiple types of cancer. J Surg Oncol 1999; 71: 130-135.

34.

Farina AR , Tacconelli A. , Vacca A. , et al. Transcriptional up-regulation of matrix metalloproteinase-9 expression during spontaneous epithelial to neuroblast phenotype conversion by SK-N-SH neuroblastoma cells, involved in enhanced invasivity, depends upon GT-box and nuclear factor kappaB elements. Cell Growth Differ 1999; 10: 353-367.

35.

Bond M. , Fabunmi RP , Baker AH , et al. Synergistic upregulation of metalloproteinase-9 by growth factors and inflammatory cytokines: an absolute requirement for transcription factor NF-kappa B. FEBS Lett 1998; 435: 29-34.

36.

Rhee JW , Lee KW , Sohn WJ , et al. Regulation of matrix metalloproteinase-9 gene expression and cell migration by NF-kappa B in response to CpG-oligodeoxynucleotides in RAW 264.7 cells. Mol Immunol 2007; 44: 1393-400.

37.

McLean GW , Carragher NO , Avizienyte E. , et al. The role of focal-adhesion kinase in cancer - a new therapeutic opportunity. Nat Rev Cancer 2005; 5: 505-515.

38.

Parsons JT Focal adhesion kinase: the first ten years. J Cell Sci 2003; 116: 1409-1416.

39.

Chin L. , Tam A. , Pomerantz J. , et al. Essential role for oncogenic Ras in tumour maintenance. Nature 1999; 400: 468-472.

40.

Campbell PM , Der CJ Oncogenic Ras and its role in tumor cell invasion and metastasis. Semin Cancer Biol 2004; 14: 105-114.

41.

Feldkamp MM , Lala P. , Lau N. , et al. Expression of activated epidermal growth factor receptors, Ras-guanosine triphosphate, and mitogen-activated protein kinase in human glioblastoma multiforme specimens. Neurosurgery 1999; 45: 1442-1453.