| Objection:(1)Using Arraystar LncRNA microarray to detect the expression of Lnc RNA and mRNA in gastric cancer tissues and adjacent normal tissues,selecting the distinct expression of LncRNA DPP10-AS1,the expression of LncRNA DPP10-AS1 in four gastric cancer cell lines MGC-803,BGC-823,SGC-7901,AGS and normal gastric mucosa epithelial cells(GES-1)were measured furtherly in the group of gastric cancer(GC)tissues and adjacent noncancerous tissues(ANTs);(2)To investigate the expression levels of Lnc RNA DPP10-AS1 adjacent gene DPP10 in gastric cancer tissues and adjacent noncancerous tissues,four gastric cancer cell lines and normal gastric mucosal cells were determined;(3)To explicit the correlation between LncRNA DPP10-AS1 and adjacent gene DPP10 expression levels in gastric cancer tissues and their expression levels in gastric cancer cells lines and normal gastric mucosal cells were consistent;(4)To explore the correlation between LncRNA DPP10-AS1 and the clinical pathological data of 76 gastric cancer patients for providing their clinical significance;(5)Through the Real-time PCR detection in gastric cancer tissues several classical molecular signaling pathways in the key gene expression levels,further screening analysis of LncRNA DPP10-AS1 on regulation of gastric cancer may be involved in signaling pathways and molecular mechanism of concrete.Method:(1)Using Arraystar LncRNA microarray detection of gastric cancer and cancer adjacent mRNA differentially expression LncRNA and adjacent mRNA,The LncRNA DPP10-AS1 was significantly lower than that of the adjacent noncancerous tissues,and we finally decide the LncRNA DPP10-AS1 as the target LncRNA;(2)The expression levels of LncRNA DPP10-AS1 in 76 patients with gastric cancer were further verified by Real-time PCR,gastric cancer cell lines MGC-803,BGC-823,SGC-7901,AGS and normal gastric mucosal cells GES-1 were also different;(3)Through the Real-time detection further gastric cancer tissue and adjacent noncancerous tissues,gastric cancer cell lines and normal gastric mucosal cells of Lnc RNA DPP10-AS1 adjacent gene DPP10 expression difference,and contrast analysis of LncRNA DPP-AS1 and its adjacent DPP10 in gastric cancer tissues and adjacent noncancerous tissues related;(4)The clinicalpathological datas of 76 gastric cancer patients were collected,and then the correlation between LncRNA DPP10-AS1 and clinical data,as well as the relevant clinical significance was analyzed;(5)Through the Real-time PCR to detect the expression of several classical associated molecular in signaling pathway in gastric cancer tissues,and furtherly screen the mechanism of LncRNA DPP10-AS1 which participates the development of gastric cancer by involving some kind of signaling pathways.Result:(1)The expression level of LncRNA DPP10-AS1 in gastric cancer tissues was significantly lower than that of adjacent noncancerous tissues(p<0.05),and the expression level of LncRNA DPP10-AS1 in gastric cancer cells was significantly lower than that of normal gastric mucosal epithelial(p<0.05);(2)The expression level of DPP10,as the adjacent gene of LncRNA DPP10-AS1,is markedly lower than that of adjacent noncancerous tissues(p < 0.05),while DPP10 expression level in gastric cancer cell lines was down-regulated compared with that in normal gastric mucosal epithelial cells(p<0.05);(3)The expression levels of LncRNA DPP10-AS1 and adjacent gene DPP10 were positively correlated in gastric cancer tissues and adjacent noncancerous tissues(R=0.901,p<0.001);The expression of DPP10 in gastric cancer cell lines was lower than that of normal gastric mucosal epithelial cells(p<0.05);(4)Through the analysis of clinical pathological data,the expression level of DPP10-AS1 was correlated with patients,Lymph node metastasis(p<0.05),and there is no relationship among the expression level of LncRNA DPP10-AS1 and patients,smoking,the size of tumor,the depth of invasion,Tumor TNM stage(p>0.05);(5)Two key genes MEK and ERK expression of MAPK/ERK signaling pathway in low expression LncRNA DPP10-AS1 group was obviously higher compared with the high expression group(p<0.05),which indicates LncRNA DPP10-AS1 may mediated the occurrence,development and distant metastasis of gastric cancer through MAPK/ERK signal pathway.Conclusion:(1)The expression level of Lnc RNA DPP10-AS1 and its adjacent gene DPP10 were significantly lower in both gastric cancer tissues and gastric cancer cell lines,and the expression of LncRNA DPP10-AS1 was positive correlated with DPP10 in gastric cancer tissues.The result showed that LncRNA DPP10-AS1 might involved in the process of biological activity regulation as DPP10 targeted;(2)The expression level of LncRNA DPP10-AS1 is closely related to Lymph node metastasis in gastric cancer patients,which signifies LncRNA DPP10-AS1 might has the prognosis significance of guiding for gastric cancer patients;(3)MAPK/ERK signaling pathway of two key genes MEK and ERK expression in low expression LncRNA DPP10-AS1 group increased significantly,which prompts LncRNA DPP10-AS1 may down regulates by MAPK/ERK signaling pathway,and LncRNA DPP10-AS1 may also be mediated by MAPK/ERK signaling pathway which involved in the occurrence,development and distant metastasis of gastric cancer. |
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