[Effect of calmodulin antagonist EBB on invasion of human fibrosarcoma cell HT1080].
OBJECTIVE
To investigate the potential effect of EBB, a calmodulin antagonist, on invasion of human fibrosarcoma cells HT1080.


METHODS
The antitumor effect of EBB was assessed by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Activities of matrix metalloproteinase (MMP)-2 and MMP-9 were measured by Zymogrophy analysis. The mRNA levels, of MMP-2, MMP-9, and tissue inhibitor of metalloproteinases (TIMP)-1 were evaluated by reverse transcriptionpolymerase chain reaction (RT-PCR). Transwell chamber assay was applied to measure the effect of EBB on the invasion of HT1080 cells.


RESULTS
Calmodulin antagonist EBB inhibited the proliferation of HT1080 cells with an IC50 of (8.2 +/- 1.2) microg/ml. EBB down-regulated the activities of MMP-2 and MMP-9, and down-regulated the mRNA levels of MMP-2 and MMP-9, while up-regulated the mRNA levels of TIMP-1. The invasive ability of HT1080 cells was decreased to (31.13 +/- 2.265)%, (59.91 +/- 2.566)%, and (71.58 +/- 0.5960)% after exposure of the cells with 2, 5, and 10 microg/ml EBB, respectively.


CONCLUSIONS
Treatment with calmodulin antagonist EBB is effective in suppressing tumor invasion. The possible mechanism is the down-regulation of MMPs.
Pan
Bing
B
State Key Laboratory of Experimental Hematology, Institute of Hematology, CAMS and PUMC, Tianjin 300020, China.
Zhou
Yuan
Y
Qi
Jing
J
Xiong
Dong-sheng
DS
Liu
Jie-wen
JW
Qi
Shu-ling
SL
Cheng
Yan-hong
YH
Yang
Chun-zheng
CZ
Zhu
Hui-fang
HF
D000970:Antineoplastic Agents / Q000494:pharmacology; D044182:Benzylisoquinolines / Q000494:pharmacology*; D002147:Calmodulin / Q000037:antagonists & inhibitors*; D045744:Cell Line, Tumor; D015536:Down-Regulation; D005354:Fibrosarcoma / Q000473:pathology*; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis* / Q000235:genetics; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis / Q000235:genetics; D009361:Neoplasm Invasiveness; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics
D000970:Antineoplastic Agents; D044182:Benzylisoquinolines; D002147:Calmodulin; D012333:RNA, Messenger; C050387:O-(4-ethoxybutyl)berbamine; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9