Effect of a nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester on invasion of human colorectal cancer cell line SL-174T.
OBJECTIVE
To investigate the effect and mechanism of action of the nitric oxide synthase (NOS) inhibitor NG-nitro-L-arginine methyl ester (L-NAME) on invasion and metastasis of human colorectal cancer cell line SL-174T.


METHODS
Human colorectal cancer cell line SL-174T was cultured and treated separately with four different dosages of L-NAME for 72 h. Nitric oxide (NO) production was measured with Griess reagent. The effect of L-NAME on invasion and migration of SL-174T cells were evaluated by using Transwell chambers attached with polycarbonate filters and reconstituted basement membrane (Matrigel). RT-PCR was performed to determine the mRNA levels of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor metalloproteinase-2 (TIMP-2).


RESULTS
L-NAME could significantly inhibit NO production of SL-174T in a dose-dependent manner. After being treated for 72 h with 0.2, 0.4, 0.8, and 1.0 mmol/L L-NAME, respectively, the ability of the L-NAME treated SL-174T cells to invade the reconstituted basement membrane decreased significantly (t = 8.056, P< 0.05; t = 14.467, P< 0.01; t = 27.785, P< 0.01; and t = 29.405, P< 0.01, respectively) and the inhibition rates were 10.29%, 19.62%, 34.08%, and 42.23%, respectively. Moreover, L-NAME could inhibit migration of SL-174T cells, and the inhibition rates were 20.76%, 24.95%, 39.43%, and 46.85% for L-NAME at 0.2, 0.4, 0.8, and 1.0 mmol/L, respectively (t= 15.116, P< 0.01). In addition, after treatment with L-NAME, expression of MMP-2 mRNA was significantly decreased (t = 71.238, P< 0.01) and that of TIMP-2 mRNA was markedly increased (t = -13.020, P< 0.01).


CONCLUSIONS
L-NAME exerts anti-invasive and anti-metastatic effects on SL-174T cell line via downregulating MMP-2 mRNA expression and upregulating TIMP-2 mRNA expression.
Yu
Li-Bo
LB
Department of General Surgery, Tumor Hospital Affiliated Harbin Medical University, Harbin 150040, Heilongjiang Province, China.
Dong
Xin-Shu
XS
Sun
Wen-Zhou
WZ
Zhao
Dong-Lu
DL
Yang
Yue
Y
D045744:Cell Line, Tumor / Q000187:drug effects*; D002465:Cell Movement / Q000187:drug effects; D015179:Colorectal Neoplasms; D004305:Dose-Response Relationship, Drug; D005260:Female; D006801:Humans; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology*; D009361:Neoplasm Invasiveness; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors*; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics / Q000378:metabolism
D019716:Tissue Inhibitor of Metalloproteinase-2; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D020778:Matrix Metalloproteinase 2; D019331:NG-Nitroarginine Methyl Ester