The expression of matrix metalloproteinase-11 gene in gastric cancer cell lines and its correlation with transcription factor Sp1 / 中国医师杂志

ABSTRACT

Objective To explore the expression of matrix metalloproteinase-11 (MMP-11) gene in gastric cancer cells and its correlation with transcription factor Sp1.Methods T In the present study,the expression of the MMP-11 was studied and the importance of Sp1 in the expression of the MMP-11 was proved with reverse transcription polymerase chain reaction (RT-PCR),differential PCR,Western-Blot,and DNA sequencing analysis in 10 kinds of the gastric cancer cell lines.Results MMP-11 cDNA was amplified with RT-PCR and differential PCR.In 34 cycle,the MMP-11/β-actin of SNU16,BGC823,MKN45,SGC7901,MGC803,SNU5 and PAMC82 were 3.32 ±0.12,2.22 ±0.11,1.41 ±0.12,1.35 ± 0.05,1.19 ± 0.05,0.97 ± 0.05,and 0.79 ± 0.05 (F =371.54,P ＜ 0.001).DNA sequencing analysis and BLAST showed the MMP11/β-actin of SNU16 DNA was 2 ～2.5 times as much as that of others.This result indicated that MMP-11 gene was amplified on the SNU16 DNA.Differential PCR and DNA sequencing analysis were carried out to amplify the MMP-11 gene in the SNU16 cell.Toward a better understanding of transcription of the MMP-1 1 gene,bioinformics technology was used to demonstrate the existence of two GC-boxes of the 5'-flanking region of MMP-11 DNA.Sp1 protein and MMP-11 protein were over-expressed in tumor cells and tended to have correlation with them (r =0.951,P ＜ 0.05).Conclusions The present results described the expressions of MMP-11 and Sp1 in gastric cancer cells.Moreover and clarified the correlation between MMP-11 and Sp1.It will establish the basic theory in order to confirm further whether Sp1 is a significant promoter of the 5'-flanking region of MMP-11 DNA.