The expression of matrix metalloproteinase-11 gene in gastric cancer cell lines and its correlation with transcription factor Sp1

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-11 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. Key words: Stomach neoplasms/ME; Matrix metalloproteinase 11/ME; Sp transcription factors/ME