Study On The Role Of MMP-11 In The Proliferation And Invasion Of Gastric Cancer Cells And The Mechanism

Gastric cancer(GC)is one of the most common malignancies.Its incidence is the fourth most common cancer incidence in the world and mortality is the second most common cancer in the world.When majority of patients visit their physicians,they are already in advanced stages with poor treatment and poor prognosis,which seriously endanger the quality of life of the citizens.Therefore,it is a major public health problem to be solved urgently.The development of GC is a complex biological process with multiple factors,multiple steps and multiple genes.At present,the molecular mechanism of GC is not clear.Therefore,exploring molecular biological mechanism of GC cell proliferation and invasion,and looking for biological markers associated with GC proliferation and invasion has been one of the important research projects in the GC basic and clinical research.The expression of matrix metalloproteinase family(MMPs)is closely related to the occurrence,invasion and prognosis of many tumors.They not only participate in degradation of extracellular matrix(ECM),affect the adhesion and motion of tumorcells,but also are related with the occurrence,invasion and metastasis of the tumor.MMP-11 belongs to the stromelysin of MMP family and there is not much research on it.As for action mechanism,MMP-11 may participate in and influence the occurrence and development of the tumor(including anti-apoptosis,cell proliferation,cell adhesion,angiogenesis)via IGF-1 way.Insulin-like growth factor(IGF)family plays an important role in the occurrence and development of malignant tumors.Among them,IGF-I is a kind of important mitogen,mainly play its biological role through insulin-like growth factor-? receptor(IGF-1R).IGF-1 is a single chain consisting of 70 amino acids.It consists of three disulfide bonds and four structural domains with a molecular weight of 7649 k Da.It plays an important role in regulating cell growth and metabolism.To explore the role and mechanism of MMP-11 in the occurrence and development of gastric cancer,,in the second section,clinical gastric cancer samples and gastric cancer cell lines SGC7901 were taken as research objects.The expression of MMP-11 and IGF-1 in different stages and types of GC cells,the effects of MMP-11 and IGF-1 on the proliferation and invasion of GC cells were studied by means of immunohistochemistry,flow cytometry,fluorescence quantitative RT-PCR,western blot and other technical systems.Part? Expression of MMP-11 and IGF-1 in gastric cancer tissueObjective: This part of the experiment aims to explore the expression of IGF-1 and MMP-11 in in different stages and types of GC tissues and normal gastric mucosal tissues.Methods: We selected 202 patients with GC who were diagnosed and underwent surgical treatment.In this study,gastric mucosa with tumor margins greater than 5cm were selected as a normal control group.The GC tissue samples were taken from the medial wall of the cancer ulcer depression.The expression levels of IGF-1 and MMP-11 in intestinal type,diffuse type and different stages(?,?,?,? stage)of GC tissues and the adjacent cancer tissues were measured by q RT-PCR.The protein expression of IGF-1 and MMP-11 in intestinal type,diffuse type and different stages(?,?,?,? stage)of GC tissues and the adjacent tissues were determined by using the western blotting.Results: By q RT-PCR and western blotting,we found MMP-11 expression in intestinal type and diffuse type of GC tissues was significantly increased compared with the adjacent tissues.MMP-11 expression in intestinal type of GC tissue was much higher than that in diffuse type of GC tissue.MMP-11 level increased with the clinical staging of GC,the expression in ? stage was significantly higher than?stage.MMP-11 level in ? stage GC tissue was significantly higher in ?and ?period.By q RT-PCR and western blotting,we found IGF-1 expression in intestinal type and diffuse type of GC tissues was significantly increased compared with the adjacent tissues.IGF-1 expression in intestinal type of GC tissue was much higher than that in diffuse type of GC tissue.IGF-1 level increased with the clinical staging of GC,the expression in ? stage was significantly higher than?stage.IGF-1 level in ? stage GC tissue was significantly higher in ?and ? period.Conclusion: The occurrence of GC can raise the expression level of MMP-11 and IGF-1 and the expression of MMP-11 and IGF-1 was related with the classification and staging of GC cells.It suggested that MMP-11 and IGF-1 may play an important inhibitory effect on the development and invasion of GC.Part? Action mechanism of MMP-11 intervention in IGF-1in gastric cancer cellsObjective: This part of the experiment aims to explore the influence of MMP-11 on the expression of IGF-1 and the action mechanism.Methods: We purchased GC cells SGC-7901 and randomly divided them into 5groups: rh-MMP-11 intervention(25 ? 50 ? 100 ng/ml rh-MMP-11 was added in SGC-7901 cell medium),NT157 intervention(5 ? 10 ?M NT157 was added in SGC-7901 cell medium after adding 50 ng/ml rh-MMP-11),piceatannol intervention(10?20 ?M piceatannol was added in SGC-7901 cell medium after adding 50 ng/ml rh-MMP-11),si-STAT3 intervention(si-STAT3 plasmid was transfected to SGC-7901 cell medium after adding 50 ng/ml rh-MMP-11)and control group.The m RNA levels and protein expression of IGF-1 in each group were determined by q RT-PCR and western blotting,respectively.Results: By q RT-PCR,we found the m RNA expression of IGF-1 in SGC-7901 cells increased significantly with the increase of MMP-11 concentration,and the protein expression of STAT3,JAK1 and IGF-1 significantly increased.The expression level of IGF-1 was significantly decreased after adding different concentrations of NT157 or percutaneous.In addition,with the increase of the concentration of NT157 or percutaneous,the m RNA level of IGF-1 in SGC-7901 cells decreased significantly,and the decrease degree of STAT3,JAK1 and IGF-1R protein expression also increased.In addition,the m RNA expression of IGF-1 in SGC-7901 cells was significantly after transfection with si-STAT3 based on the MMP-11 intervention culture.Conclusion: MMP-11 could promote the phosphorylation of STAT3,JAK1 and IGF-1R.NT157 could decrease the phosphorylation of STAT3,JAK1 and IGF-1R.MMP-11 can activate the JAK1/STAT3 signaling pathway by inducing IGF-1expression.Part ? Functional mechanism of MMP-11 and IGF-1 in the proliferation and invasion of gastric cancer cellsObjective: This part of the experiment aims to explore the effects of IGF-1 and MMP-11 on the proliferation and invasion of GC cells.Methods: We purchased GC cells SGC-7901 and randomly divided them into 5groups: rh-IGF-1 intervention(25?50?100 ng/ml rh-IGF-1 was added in SGC-7901 cell medium),NT157 intervention(10 ?M NT157 was added in SGC-7901 cell medium after adding 50 ng/ml rh-IGF-1),piceatannol intervention(20 ?M piceatannol was added in SGC-7901 cell medium after adding 50 ng/ml rh-IGF-1),si-STAT3 intervention(si-STAT3 plasmid was transfected to SGC-7901 cell medium after adding 50 ng/ml rh-IGF-1)and control group.In addition,some GC cells were randomly divided them into 5 groups: rh-MMP-11 intervention(1050 ng/ml rh-IGF-1was added in SGC-7901 cell medium),NT157 intervention(10 ? M NT157 was added in SGC-7901 cell medium after adding 50 ng/ml rh-MMP-11),piceatannol intervention(20 ?M piceatannol was added in SGC-7901 cell medium after adding50 ng/ml rh-MMP-11),si-STAT3 intervention(si-STAT3 plasmid was transfected to SGC-7901 cell medium after adding 50 ng/ml rh-MMP-11)and control group.MTT method was used to determine the proliferation situation of GC cells in each group and Transwell invasion experiment was used to determine the invasion situation of GC cells in each group.Results: The proliferation and invasion capacity of SGC-7901 cells increased significantly after IGF-1 intervention.The proliferation and invasion capacity is stronger with the intervention time increase.However,the enhancement degree decreased after NT157 intervention,piceatannol intervention or si-STAT3 transfection.The proliferation and invasion capacity of SGC-7901 cells increased significantly after MMP-11 intervention.The proliferation and invasion capacity is stronger with the intervention time increase.However,the enhancement degree decreased after NT157 intervention,piceatannol intervention or si-STAT3 transfection.Conclusion: MMP-11 can promote the proliferation and invasion of GC cells by stimulating IGF-1 expression.