| Objective: Gastric cancer is one of the high morbidity and mortality of cancer in the world.China is the high incidence area of gastric cancer,the incidence rate of gastric cancer in China is higher than the world average level.The exact molecular components of the pathogenesis of gastric cancer have not been fully elucidated.Recent research shows that a class of non coding protein RNA called long non coding RNA(Lnc RNA)is involved in the pathogenesis of many human diseases,including cancer.In this study,LncRNA ADAMTS9-AS1 was selected as the object of study from the results of Arraystar LncRNA Microarray V3.0.To further study the expression of LncRNA ADAMTS9-AS1 in a large number of gastric cancer tissue samples.The correlation between clinical pathological factors and expression of LncRNAADAMTS9-AS1 was analyzed.Meanwhile,to analyze the correlation between expression of LncRNA ADAMTS9-AS1 and immunohistochemical markers.Inhibition of LncRNA ADAMTS9-AS1 expression in gastric adenocarcinoma cells,study its effect on function of gastric adenocarcinoma cells.The possible downstream molecules analyzed by bioinformatics is ADAMTS9.Preliminary study on the correlation between ADAMTS9-AS1 LncRNA and ADAMTS9.This study lays a foundation for the further understanding of the role of Lnc RNA ADAMTS9-AS1 in the pathogenesis of cancer.Meanwhile to search for gastric adenocarcinoma diagnostic markers and therapeutic targets to provide the basis.Methods:(1)We selected a representative of the 6 gastric adenocarcinoma tissues and adjacent normal tissues,it was detected on differential expression of LncRNAbetween 6 gastric adenocarcinoma tissues and adjacent normal tissues by Arraystar LncRNA Microarray V3.0.LncRNA ADAMTS9-AS1 was selected as the object of study from result of Microarray.(2)It was detected on expression of LncRNA ADAMTS9-AS1 between 132 gastric adenocarcinoma tissues and adjacent normal tissues by qRT-PCR technology.The consistency of the results of the above two experiments will be verified.The correlation between clinical pathological factors and expression of LncRNA ADAMTS9-AS1 was analyzed.Meanwhile,to analyze the correlation between expression of Lnc RNAADAMTS9-AS1 and immunohistochemical markers.(3)After LncRNA ADAMTS9-AS1 was knockdowned by siRNA,using CCK-8assay,colony formation assay,Transwell cell migration and invasion assay,Cell apoptosis,we studied gastric adenocarcinoma cell biological function changes.(4)The possible downstream molecules of LncRNA ADAMTS9-AS1 analyzed by bioinformatics analysis was ADAMTS9.The expression of ADAMTS9 in gastric adenocarcinoma tissue were detected by qRT-PCR assay.To analysis of the correlation between the expression of LncRNAADAMTS9-AS1 and ADAMTS9.Results:(1)Result of lncRNA gene chip: In 6 patients with gastric cancer tissues and adjacent normal tissue contrast,expression of LncRNA ADAMTS9-AS1 were up-regulating,fold chang is 4.92 fold.(2)Result of qRT-PCR assay in 132 gastric adenocarcinoma tissues:The expression of LncRNA ADAMTS9-AS1 were up-regulating in 132 gastric adenocarcinoma tissues with paired normal tissue as control.Up rate was 66%,fold chang is 6.26 fold(P<0.05).Statistics show that it are correlated between expression of LncRNAADAMTS9-AS1 and tumor size,tumor invasion,vessel invasion,VEGF.(3)Result of Functional study of LncRNAADAMTS9-AS1 in gastric adenocarcinoma cell: The expression of LncRNA ADAMTS9-AS1 in 4 cell lines of gastric adenocarcinoma(BGC-823,SGC-7901,MGC-803,AGS)were up-regulating,with GES-1 control.After two different sites of siRNA knockdown of LncRNA ADAMTS9-AS1 in 2 cell lines of gastric adenocarcinoma,We found that cellproliferation,colony formation,migration and invasion were significantly inhibited.(4)Preliminary exploration results of downstream molecules of LncRNA ADAMTS9-AS1 : The justice chain of LncRNA ADAMTS9-AS1 is ADAMTS9 gene.The expression of ADAMTS9 were down regulation in gastric adenocarcinoma tissues with paired normal tissue as control.Statistics show that it are correlated between down-regulating expression of ADAMTS9 and tumor size,vessel invasion,VEGF.Linear regression analysis showed that ADAMTS9 expression was negatively correlated with LncRNA ADAMTS9-AS1 expression,r=0.754,R2=0.568,P<0.05.Conclusions:.Result of Arraystar Lnc RNA Microarray V3.0,gastric adenocarcinoma tissue qRT-PCR verification and gastric adenocarcinoma cell lines verification in the expression of LncRNA ADAMTS9-AS1 is consistent.Their result were that the expression of LncRNA ADAMTS9-AS1 in gastric adenocarcinoma were up-regulating.Result of Functional study of LncRNAADAMTS9-AS1 in gastric adenocarcinoma cell is that decreased LncRNA ADAMTS9-AS1 can inhibit gastric adenocarcinoma cell proliferation ability,coloning ability and invasion ability.Therefore,upregulation of LncRNAADAMTS9-AS1 promotes tumor proliferation and invasion.The statistical analysis found that it was correlated between the up-regulated expression of LncRNAADAMTS9-AS1 and tumor size,tumor invasion,vascular invasion,higher expression of VEGF in gastric adenocarcinoma patients.Meanwhile,Our research shows that the expression of ADAMTS9 as justice chain of LncRNAADAMTS9-AS1 is down-regulating in gastric adenocarcinoma.Statistics show that it are correlated between down-regulated expression of ADAMTS9 and tumor size,vessel invasion,higher expression of VEGF in gastric adenocarcinoma patients.It was basically the same statistical analysis result in clinical pathological factors and immunohistochemical markers in LncRNAADAMTS9-AS1 and ADAMTS9.Linear regression analysis revealed that the expression level of LncRNA ADAMTS9-AS1 and ADAMTS9 was negatively correlated.Combined with the literature and experimental results,we speculate that up-regulated LncRNAADAMTS9-AS1 can negative regulate ADAMTS9,and then down regulate ADAMTS9 downstream molecules such as VEGF by the signal transduction pathway of ADAMTS9.play a role in regulation and control.The final result of this regulation is to promote the proliferation and invasion of tumor. |
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