Modulation of cell growth and matrix metalloproteinase-2 activation of oral squamous cell carcinoma as a function of culture condition with type I collagen.
OBJECTIVE
Oral squamous cell carcinoma (OSCC) is characterized by local invasiveness Matrix metalloproteinase-2 (MMP-2) is a key enzyme involved in local invasiveness. Type I collagen functions as a modulator of cellular function, including MMP-2 activation; additionally, it serves as a barrier against tumor invasion. This study was designed to analyze the effect of type I collagen on tumor growth and MMP-2 activation of OSCC cell lines as a function of culture conditions.


METHODS
Two OSCC cell lines (SAS, OSC-19) were cultured under three conditions: conventional monolayer culture on plastic dishes (MP); two-dimensional culture on type I collagen gel (2D); and three-dimensional culture within a type I collagen gel (3D). MMP-2 activity was determined by means of gelatin zymography and invasive cell growth was assessed morphologically. The inhibitory effect of BB-2516 (marimastat) on SAS and OSC-19 cell growth was examined. Additionally, the effect of tissue inhibitor of MMP-1 (TIMP-1) and TIMP-2 on SAS cell growth was investigated.


RESULTS
The most intense MMP-2 activation was evident in the 3D cases in comparison with MP and 2D in both SAS and OSC-19. MMP-2 activation was correlated with cell growth. MMP-2 activation and cell growth were inhibited by BB-2516 in a dose-dependent manner. Dose-dependent inhibitory effects of TIMP-2, but not TIMP-1, against MMP-2 activation were observed in SAS.


CONCLUSIONS
Three-dimensional type I collagen gel culture is a valuable method for monitoring both MMP-2 activation and invasive cell growth. Membrane type 1 matrix (MT1)-MMP supposedly plays key roles in MMP-2 activation and cell growth in 3D. Thus, this system would be useful for evaluating the inhibitory effect of molecules against MT1-MMP.
Kinsenn
Hidenori
H
Department of Otolaryngology, School of Medicine, Cancer Research Institute, Kanazawa University, Kanazawa, Japan.
Sato
Hiroshi
H
Furukawa
Mitsuru
M
Yoshizaki
Tomokazu
T
D002294:Carcinoma, Squamous Cell / Q000201:enzymology* / Q000473:pathology*; D018929:Cell Culture Techniques; D002455:Cell Division; D045744:Cell Line, Tumor; D024042:Collagen Type I / Q000502:physiology*; D004789:Enzyme Activation; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors; D009062:Mouth Neoplasms / Q000201:enzymology* / Q000473:pathology*; D009361:Neoplasm Invasiveness
D024042:Collagen Type I; D061965:Matrix Metalloproteinase Inhibitors; D020778:Matrix Metalloproteinase 2