Abstract
Purpose
Cell-matrix adhesive interaction has an important role in the invasive process of tumor cells, and integrins are the major receptors mediating cell-matrix adhesion. The current study is to investigate the modulation of basement membrane (BM) proteins, especially collagen IV (C IV), laminin (LN), and fibronectin (FN) in the invasive processes of human hepatocellular carcinoma (HCC) cells in vitro, and to reveal the roles of β1 integrins and RGD-containing oligopeptide in the cell-matrix interaction.
Methods
Static adhesion assay was performed to study the rates of adhesion of MHCC97-H cells, treated or untreated with anti-β1 (2 μg ml–1) and GRGDS, to C IV (50 μg ml–1), LN (50 μg ml–1) or FN (50 μg ml–1). Gelatin zymography was used to detect the secretion of MMPs in the conditioned medium of MHCC97-H cells incubated 24 h by C IV, LN or FN, and interactions with anti-β1 and GRGDS. Transwell chamber assay was used to investigate the influence of C IV, LN or FN, interacting with anti-β1 and GRGDS, on the cellular mobility of MHCC97-H cells.
Results
Compared with blank control group, MHCC97-H cells showed significantly higher rates of adhesion to C IV, LN, and FN. Pretreatment with anti-β1 could suppress adhesion to C IV, LN or FN, but GRGDS inhibited adhesion to FN (P<0.05) only. LN and FN could stimulate the secretion of MMPs by MHCC97-H cells cultured in vitro, especially MMP-9 and its activated type. Treatment with anti-β1 could partly counteract the effects of LN and FN. GRGDS could prominently induce the secretion of MMPs, but the effect could be inhibited by pretreatment of anti-β1. The results of Transwell chamber assay showed that LN, FN, and GRGDS could increase the number of tumor cells penetrating the microporous membrane, but the data of C IV did not reach significance. The effects were partly counteracted by anti-β1.
Conclusion
BM proteins play an active role in the invasive process of human hepatocellular carcinoma cells. Integrin β1 is an important molecule which mediates the cell-matrix adhesive interaction of tumor cells. RGD-containing peptides competitively combine with the binding site of integrin β1, and the effects of FN are RGD sequence-dependent.
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References
Begum NA, Mori M, Matsumata T, Takenaka K, Sugimachi K, Barnard GF (1995) Differential display and integrin a6 messenger RNA overexpression in hepatocellular carcinoma. Hepatology 22:1447–1455
Berry MG, Goode AW, Puddefoot JR, Vinson GP, Carpenter R (2003) Integrin beta1-mediated invasion of human breast cancer cells: an ex vivo assay for invasiveness. Breast Cancer 10:214–219
Cachau RE, Serpersu EH, Mildvan AS, August JT, Amzel LM (1989) Recognition in cell adhesion. A comparative study of the conformations of RGD-containing peptides by Monte Carlo and NMR methods. J Mol Recognit 2:179–186
Dedhar S, Hannigan GE (1996) Integrin cytoplasmic interactions and bi-directional transmembrane signaling. Curr Opin Cell Biol 8:657–669
Engbring JA, Kleinman HK (2003) The basement membrane matrix in malignancy. J Pathol 200:465–470
Felding-Habermann B (2003) Integrin adhesion receptors in tumor metastasis. Clin Exp Metastasis 20:203–213
Fleischmajer R, Perlish JS, MacDonald ED, Schechter A, Murdoch AD, Iozzo RV, Yamada Y (1998) There is binding of collagen IV to beta 1 integrin during early skin basement membrane assembly. Ann N Y Acad Sci 857:212–227
Flug M, Kopf-Maier P (1995) The basement membrane and its involvement in carcinoma cell invasion. Acta Anat 152:69–84
Fujii, Nishikawa N, Komazawa H, Suzuki M, Kojima M, Itoh I, Obata A, Ayukawa K, Azuuma I, Saiki I (1998) A new pseudopeptide of Arg-Gly-Asp (RGD) with inhibitory effect on tumor metastasis and enzymatic degradation of extracellular matrix. Clin Exp Metastasis 16:94–104
Haier J, Nasralla M, Nicolson GL (1999) Different adhesion properties of highly and poorly metastatic HT-29 colon carcinoma cells with extracellular matrix components: role of integrin expression and cytoskeletal components. Br J Cancer 80:1867–1874
Hemler ME, Mannion BA, Berditchevski F (1996) Association of TM4SF proteins with integrins: relevance to cancer. Biochem Biophys Acta 1287:67–71
Heussen C, Dowdle EB (1980) Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Anal Biochem 102:196–202
Hida J, Matsuda T, Kitaoka M, Machidera N, Kubo R, Yasutomi M (1994) The role of basement membrane in colorectal cancer invasion and liver metastasis. Cancer 74:592–598
Humphries MJ, Olden K, Yamada KM (1986) A synthetic peptide from fibronectin inhibits experimental metastasis of murine melanoma cells. Science 233:467–470
Hynes RO (1992) Integrins: versatility, modulation, and signaling in cell adhesion. Cell 69:11–25
Jaskiewicz K, Chasen NR (1995) Differential expression of transforming growth factor a, adhesions molecules and integrins in primary, metastatic liver tumor and in liver cirrhosis. Anticancer Res 15:559–562
Juliano RL, Varner JA (1993) Adhesion molecules in cancer: the role of integrins. Curr Opin Cell Biol 5:812–818
Kleinman HK, Koblinski J, Lee S, Engbring J (2001) Role of basement membrane in tumor growth and metastasis. Surg Oncol Clin N Am 10:329–338
Koo LY, Irvine DJ, Mayes AM, Lauffenburger DA, Griffith LG (2002) Co-regulation of cell adhesion by nanoscale RGD organization and mechanical stimulus. J Cell Sci 115:1423–1433
Li Y, Tang ZY, Ye SL, Liu YK, Chen J, Xue Q, Chen J, Gao DM, Bao WH (2001) Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97. World J Gastroenterol 7:630–636
Maheshwari G, Brown G, Lauffenburger DA, Wells A, Griffith LG (200) Cell adhesion and motility depend on nanoscale RGD clustering.J Cell Sci 113:1677–1686
Okroj M, Dobrzanska-Paprocka Z, Rolka K, Bigda J (2003) In vitro and in vivo analyses of the biological activity of RGD peptides towards Ab bomirski melanoma. Cell Mol Biol Lett 8:873–884
Oshita F, Kameda Y, Ikehara M, Tanaka G, Yamada K, Nomura I, Noda K, Shotsu A, Fujita A, Arai H, Ito H, Nakayama H, Mitsuda A (2002) Increased expression of integrin beta1 is a poor prognostic factor in small-cell lung cancer. Anticancer Res 22:1065–1070
Ozaki I, Yamamoto K, Mizuta T, Kajihara S, Fukushima N, Setoguchi Y, Morito F, Sakai T (1998) Differential expression of laminin receptors in human hepatocellular carcinoma. Gut 43:837–842
Stetler-Stevenson WG, Aznavoorian S, Liotta LA (1993) Tumor cell interactions with the extracellular matrix during invasion and metastasis. Ann Rev Cell Biol 9:541–573
Stracke ML, Liotta LA (1992) Multi-step cascade of tumor cell metastasis. In Vivo 6:309–316
Stracke ML, Murata J, Aznavoorian S, Liotta LA (1994) The role of the extracellular matrix in tumor cell metastasis. In Vivo 8:49–58
Tang ZY (2000) Surgery of hepatocellular carcinoma with special reference to studies on metastasis and recurrence. Gastroenterol Today 4:191–195
Tsuchiya Y, Sawada S, Tsukada K, Saiki I (2002) A new pseudo-peptide of Arg-Gly-Asp (RGD) inhibits intrahepatic metastasis of orthotopically implanted murine hepatocellular carcinoma. Int J Oncol 20:319–324
Wilson DF, Jiang DJ, Pierce AM, Wiebkin OW (1999) Oral cancer: role of the basement membrane in invasion. Aust Dent J 44:93–97
Yang C, Zeisberg M, Lively JC, Nyberg P, Afdhal N, Kalluri R (2003) Integrin alpha1beta1 and alpha2beta1 are the key regulators of hepatocarcinoma cell invasion across the fibrotic matrix microenvironment. Cancer Res 63:8312–8317
Yao M, Zhou XD, Zha XL, Shi DR, Fu J, He JY, Lu HF, Tang ZY (1997) Expression of the integrin a5 subunit and its mediated cell adhesion in hepatocellular carcinoma. J Cancer Res Clin Oncol 123:435–440
Zitzmann S, Ehemann V, Schwab M (2002) Arginine-glycine-aspartic acid (RGD)-peptide binds to both tumor and tumor-endothelial cells in vivo. Cancer Res 62:5139–5143
Acknowledgement
This project was supported in part by China State Key Basic Research Program Grant (G199805120) and Key Laboratory of Carcinogenesis and Cancer Invasion (Fudan University), Ministry of Education, P.R. China
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Tian, B., Li, Y., Ji, XN. et al. Basement membrane proteins play an active role in the invasive process of human hepatocellular carcinoma cells with high metastasis potential. J Cancer Res Clin Oncol 131, 80–86 (2005). https://doi.org/10.1007/s00432-004-0614-3
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DOI: https://doi.org/10.1007/s00432-004-0614-3