The Preliminary Exploration On The Correlation Between FOXM1 ,Mmp-9 And The Occurrence And Development Of Gallbladder Cancer

Objective: To detect the expression of FOXM1 and MMP-9 in Gallbladder Carcinoma tissues and to explore the relationship between the expression of FOXM1 and the biological behavious of human Gallbladder Carcinoma NOZ cells by decreasing FOXM1 gene expression by RNA interference. And to explore its significance in the occurrence, progression of Gallbladder Carcinoma.Methods: Immunohistochemisty was employed to detect the expression of FOXM1 and MMP-9 in 51 samples of Gallbladder Carcinoma tissues and 28 samples of CC,and statistical analyse the correlation between FOXM1 and MMP-9 as well as the relationship between their expression and clinic pathologial paramterers ; RT-PCR to detect the FOXM1 and MMP-9 expression in the three gallbladder cancer cell lines (GBC-SD, NOZ, SGC996), and screen out the cell line with FOXM1 and MMP-9 highly expressed;four pair of DNA template coding was synthesized against Gallbladder Carcinoma cells to reconstruct PGPU6/GFP/Neo-FOXM1 and screened out the strongest one which was transfected into NOZ cells with FOXM1 and MMP-9 highly expressed . RT-PCR, Western blot to detect the endogenous expression of FOXM1 in the FOXM1 siRNA plasmid / NOZ cells, RT-PCR and immunocytochemistry staining to detect the endogenous expression of MMP-9 in the FOXM1 siRNA plasmid / NOZ cells; MTT and Transwell were used to observe the change in the proliferation and invasion ability of NOZ cells. Results: The positive rates of FOXM1 and MMP-9 in the Gallbladder Carcinoma tissues were higher than in the CC tissues (52.9% and 76.5%vs. 10.7% and 7.1% , P≤0.001). The expression of FOXM1 in the Gallbladder Carcinoma tissues was significantly correlated with the clinical stage,the differentiation,lymph node metastasis ( P =0.007,0.008,0.019),and the expression of MMP-9 in the Gallbladder Carcinoma tissues was significantly correlated with the clinical stage,lymph node metastasis ( P =0.001,0.017). The expression of FOXM1 was positively correlated with the expression of MMP-9 in Gallbladder Carcinoma tissues(r =0.043, P=0.003). RT-PCR results showed that gallbladder carcinoma cell lines( GBC-SD, NOZ, SGC996 )have high expression of FOXM1 and MMP-9 and the expression in these was not statistically significant (P <0.05), and NOZ cell line was elected as a pilot cell line;RT-PCR and Western blot analyses demonstrated that PGPU6/GFP/Neo-FOXM1 could down-regulate the expression of significantly inhabit the expression of FOXM1 to 74.32% and 82.18% respectively in NOZ cells ,and also down-regulate the expression of MMP-9 in NOZ cells;MTT and Transwell results showed that PGPU6/GFP/Neo- FOXM1 could significantly inhibit the proliferation of NOZ cells and reduce the invasive cell number to(31.4±4.037).Conclusion: FOXM1 and MMP-9 in human gallbladder carcinoma tissues showed a high expression ,and were relected with clinicopathological parameters as well as the occurrence and progression of Gallbladder Carcinoma; PGPU6/GFP/Neo-FOXM1 could significantly inhibit the expression of FOXM1, suppress the growth and invasion of NOZ cells and low down the expression of MMP-9. The invasion and metastasis of human gallbladder cancer by FOXM1 may be related to increase MMP-9 expression.