The Preliminary Analysis Of Exosomal CircRNA Expression Profile In Lymph Node Metastasis Of Esophageal Squamous Cell Carcinoma

Objective:Esophageal carcinoma is one of the most common malignant tumors of digestive tract,which has serious influence on human health and quality of life.Metastasis is the primary cause of the death for esophageal carcinoma.Previous studies indicated that metastasis affect the survival rate of patients with esophageal cancer significantly.Because a considerable amount of circRNA exist in plasma exosomes which mostly are evolutionally conserved,stable and tissue specific,there is a great significance that choosing exosomal circRNA as the novel biomarkers to predict cancer prognosis.The study analyzes the expression profile of exosomal circRNA in esophageal squamous cell carcinoma?ESCC?,uses molecular biology methods,screens characteristic circRNA associated with Lymph node metastasis in esophageal carcinoma Exploring the potential of exosomal circRNA as biomarkers for ESCC,provides population and laboratory evidence for non-invasive prediction of lymph node metastasis and prognosis of ESCC,and provides clues for elucidating the mechanisms of metastasis and recurrence of ESCC.Methods: We choose 4 cases of ESCC with metastasis,4 cases of ESCC without metastasis,and 4 cases of benign diseases without systemic inflammation,total 16 plasma as samples.Then we demonstrated the difference of the expression of circRNA by microarray analysis between the patients with ESCC before and after surgical treatment,ESCC with metastasis and without metastasis,and ESCC with metastasis and benign diseases without systemic inflammation.The intersection of exosomal circRNA of the three comparison group is used as the results of the initial screening.We detected the expression levels of exosomal circRNA by 2 stages real-time PCR in 69 ESCC patients in order to confirm the results of the preliminary screening of the microarray analysis.Using survival analyze to investigate relationship between the expression levels of exosomal circRNA and prognosis in ESCC patients following surgical resection.Bioinformatics analyze is using KOBAS annotation circRNA in pathways,diseases and functional.Predict possible target mi RNAs,and speculate the possible mechanism of exosomal circRNA.Results: The result of the microarray analysis showed that 106 circRNA in the three comparison groups which are significantly correlated with metastasis in ESCC carcinoma patients especially lymph node metastasis,up express circRNA 50?P<0.05 and FC>2?,down express circRNA 56?P<0.05 and FC<0.5?.These 106 circRNA are considered associated with lymph node metastasis in ESCC.The expression of has_circRNA₀026611 in ESCC with lymph node metastasis was significantly increased by real-time q PCR?z=-3.205,P<0.01?.Survival analyze shows that high expression level of has_circRNA₀026611??1.696?is a significant adver progeostic factor of progress-free survival?P=0.034,HR=10.331?.Bioinformatics analysis shows that cancer ralated mi RNAs such as hsa-mi R-608?hsa-mi R-6743-5p and hsa-mi R-2278 might be the target of has_circRNA₀026611.KEGG pathway enrichment shows that has_circRNA₀026611 participate in tumor invasion and metastasis through micro RNAs in cancer pathway.Conclusion: The results of this study showed that the expression level of has_circRNA₀026611 increased in exosomes of plasma of ESCC with lymph node metastasis,which suggested that circRNA₀026611 has potential as a biomarker of ECSS,the up regulation of has_circRNA₀026611 in exosomes of plasma exerted potential metastasis-promoting effect in esophageal squamous cell carcinoma.And the up regulation of has_circRNA₀026611 is a significant adver prognostic factor of disease-free survival in ESCC patients following surgical resection.