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Objective:Toinvestigate the influence of MMP7 antisense oligodeoxynucleotide(ASODN)on adhesion and invasion ofhuman lung adenocarcinoma cell line A549.Methods:Phosphorothioate MMP7 ASODN was transfected to A549 cells mediated by liposome. The expression of MMP7 was examined by RT-PCR.The adhesive and invasive ability were examined by plate adhesion model and Boyden Chamber transwell assay.Results:After MMP7 ASODN was transfected,the relative optical density(ROD)of electrophoresis strip was decreased obviously(P
ABSTRACT
PURPOSE: The effects of PGE2 receptors (EP1, 2, 3, 4) on the proliferation of prostate cancer cells are still unclear. The degradation of the basement membrane by MMP-2, 7, 9 and TIMP-1, 2 is a critical point in tumor invasion and metastasis. We investigated the effects of PGE2 receptors concerning MMP and TIMP after the treatment of COX-2 inhibitors on prostate cancer cell-lines. MATERIALS AND METHODS: Two prostate cancer cell-lines, PC-3 and DU-145 cells were used in this study. RT-PCR were performed to detect the mRNA expression of EP1, 2, 3, 4, MMP-2, 7, 9 and TIMP-1, 2, MMP-7 was measured by ELISA after being treated with the selective EP2 agonist and EP4 agonist 10(-10), 10(-8), 10(-6) microM respectively. RESULTS: EP2, 3 and 4 mRNA were expressed in both cell-lines. After the NS-398 treatment, EP2 and EP4 mRNA expressions decreased in PC-3 cells. While only the MMP-7 mRNA expression decreased in PC-3 cells after NS-398 treatment, after NS-398 with selective EP2 agonist and EP4 agonist, MMP-7 mRNA expression increased. In PC-3 cells, selective EP2 agonist and EP4 agonist induced a significant dose-dependent increase in MMP-7 production in comparison to the NS-398 treatment group (control) in the conditioned ELISA medium. CONCLUSIONS: These results strongly suggest that COX-2, to some extent, contribute to prostate carcinogenesis at the EP2 and EP4 receptor, which could also be explained by increments of MMP-7 in PC-3 cells. Therefore, these findings show that selective EP inhibitor is useful in preventing specific disease progression in prostate cancer.
Subject(s)
Basement Membrane , Carcinogenesis , Cyclooxygenase 2 Inhibitors , Disease Progression , Enzyme-Linked Immunosorbent Assay , Matrix Metalloproteinase 7 , Neoplasm Metastasis , Prostaglandin-Endoperoxide Synthases , Prostate , Prostatic Neoplasms , Receptors, Prostaglandin E , RNA, Messenger , Tissue Inhibitor of Metalloproteinase-1ABSTRACT
Objective: To investigate the expressions of MMP-2 and MMP-7 between colorectal cancer and normal tissue.Methods: RT-PCR was used to determine the expressions of MMP-2 and MMP-7 in colorectal cancer and normal tissue.Results: The expression rate and level of MMP-2 and MMP-7 were higher in cancerous tissue than that in normal tissue(P
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Objective To study the effect of the recombinant matrilysin (rMMP-7) and its antisense oligonucleotide transfected by liposome (LIPO-MAON) on the proliferation of lung adenocarcinoma cell line A549 and human umbilical vein endothelial cells (HUVEC). Methods Antisense oligonucleotide of matrilysin was constructed by liposome transfection. The proliferation of HUVEC and A549 was detected by MTT assay in case of rMMP-7 or LIPO-MAON existence. The expression of MMP-7 mRNA in both HUVEC and A549 was examined by semi-quantitative RT-PCR assay. Results The proliferation of both HUVEC and A549 cell line was accelerated by high level of rMMP-7 (0.75, 1.0 ?g/ml), and the inhibited proliferation was only found in A549 cell line treated with high concentration of LIPO-MAON (7.5, 10 nmol/ml), but not in HUVEC. By RT-PCR assay, no expression of MMP-7 mRNA was found in HUVEC; however, enhanced or decreased expression of mRNA were found when A549 cell line was treated respectively with rMMP-7 or LIPO-MAON (P
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AIM:To investigate the effects of MMP-7asODN on MMP-7 expression in human stomach cancer cell line KATOIII and explore the effect of MMP-7 on cancer invasion and metastasis. METHODS: A 15mer PS-asODNs targeted against the MMP-7 mRNA were introduced into KATOIII cells by FuGENE TM 6. The levels of MMP-7 mRNA was observed by reverse transcription polymerase chain reaction (RT-PCR). The ability of the invasion was observed by Boyden Chamber invasion assay. RESULTS: The MMP-7/?-actin ration was 0.31?0.02 for the PS-asODNs treated cells which is significantly lower compared with the control and PS-sODN and PS-mODN( 1.59?0.01, 1.14?0.03,1.51?0.02, respectivly.) P
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Objective To investigate the possibility of inhibiting the malignant proliferation of tumor cells in vitro through down-regulating the expression of MMP-7 by antisense gene technology. Methods A Phosphorothioate antisense oligodeoxynucleotide (ASODN) and scrambled oligodeoxynucleotide (SCODN) against MMP-7 were respectively transfected into A549 cells mediated by liposome. The expression of MMP-7 and cell proliferation in the cells were respectively examined by immunohistochemical assay and MTT. Flow cytometry was used to detect the cell cycles. Results After A549 cells were transfected with MMP-7 ASODN, the cell number was decreased. MTT method indicated that the proliferation of A549 cells was inhibited by MMP-7 ASODN at different concentrations, with the inhibition reaching the peak at 48 h. ASODN transfection downregulated the expression levels of MMP-7 protein, and inhibited the transition period of G_ 0 /G_ 1 phase to S phase. Conclusion The artificial MMP-7 ASODN can specifically inhibit the expression of MMP-7 protein and regulate cell cycle and cell proliferation in A549 cells.