The Expression And Significance Of Hsa＿circ＿001710 In Gastric Cancinoma

Gastric cancer is a common malignancy,ranking the fifth in all incidence rate of cancer worldwide in the world at present,,and the mortality rate is the third in the world.According to the global cancer statistics in 2018,there are nearly 1 million new gastric cancer cases,and nearly783000 deaths in 2018.In China,due to the large population and the gap between the developed countries in economic development,the incidence and mortality of gastric cancer are still relatively serious,gastric cancer is the first malignant tumor in gastrointestinal cancer.In terms of its treatment,active surgical operation or postoperative adjuvant chemotherapy are common treatments,but many patients with gastric cancer still have poor prognosis,mainly because early gastric cancer is not easy to diagnose.In 2013,there are two articles on RNA published in Nature which completely broke people’s traditional understanding of RNA at that time,and quickly attracted wide attention of the whole biomedical community.At present,cric RNA has been becoming a new research hotspot in the field of RNA.In fact,circular RNA was first found in the 1970 s.Different from the traditional linear RNA,its structure is a class of closed covalent ring molecule through reverse splicing,without 5 ’ end cap and 3’ end poly A tail,which means it is not easy to be degraded by exonuclease,being highly conservative,stable and tissue characteristic.Its main functions are as follows: as the sponge of micro RNA(mi RNA),as a regulator of gene transcription,binding with RNA binding proteins,and translating its sequence into protein.Therefore,it is of great significance to screen and identify circ RNA molecules associated with gastric cancer.In this study,we screened out the target circ RNA related to gastric cancer by using Circ2 Traits database,and then analyzed whether there was differential expression in gastric cancer tissues and paracancerous tissues to explore the significance of target circrna expression in gastric cancer tissues and the possibility of being a new molecular marker in the future.Methods1.From January 2019 to June 2019,90 pairs of gastric cancer tissues and matched paracancerous tissues were collected from Ningbo clinical pathological diagnosis center,China.After about 1 cm of each sample is taken,it is immediately put in RNA fixer non-liquid nitrogen sample RNA preservation solution(bioteche,Beijing,China)at-80 ℃.2.This study included 90 pairs samples which is 5cm away from the lesions.The tumor staging system of tumor lymph node metastasis(TNM)of the international anti-cancer Federation(Eighth Edition)was used.Histological grading was assessed by the National Comprehensive Cancer Network(NCCN)guidelines for clinical practice on oncology(2020.V.1).3.FFPE RNA Kit was used to extract the total RNA that from gastric cancer tissue samples and paracancerous tissues samples.The concentration of total RNA and A260 / A280 value were detected by Ultra-micro spectrophotometer DS-11(Denovix)to determine the yield and purity.After the extraction was qualified,it was immediately stored in the ultra-low temperature refrigerator at-80 ℃.4.Synthesis of complementary DNA(c DNA)by goscript reverse transcription system.5.The c DNA template was amplified by fluorescence Quantitative Real-time reverse transcription polymerase chain reaction(QRT-PCR),and gotaq q PCR Master Mix(Promega)was used to amplify on mx3005 p real-time PCR(stratagene,La Jolla,CA,USA).,besides,the PCR reaction was performed in triplicate to eliminate random errors.The final product was sequenced to confirm wether it was the hsa＿circ＿001710.6.The expression levels of hsa＿circ＿001710 in gastric cancer and paracancerous tissues were detected by q RT-PCR,and the paired t test was used to analyze whether the expression level was statistically different.7.Data analysis was completed by SPSS 20.0(SPSS,Chicago,IL,USA)software.Chi square test was used to evaluate the relationship between the expression level of hsa＿circ＿001710 and the clinicopathological factors.Graph Pad Prism 5.0(graphpad software,La Jolla,CA,USA)was used to draw the graph.8.Constructing the Receiver Operating Characteristic(ROC),and Area Under Curve(AUC)was calculated to evaluate its diagnostic value,including sensitivity and specificity.p< 0.05 was statistically significant.Results1.Hsa＿circ＿001710 was expressed in gastric cancer.2.The expression level of hsa＿circ＿001710 in gastric cancer was significantly lower than that in adjacent tissues.(p<0.0001)3.The expression level of hsa＿circ＿001710 was correlated with tumor size(p = 0.016)and tumor differentiation(p = 0.028),but not with gender,age,degree of differentiation and so on(p >0.05).4.The area under curve(AUC)of the receiver operating characteristic curve(ROC)expressed by hsa＿circ＿001710 was 0.658,the sensitivity and specificity were 84.4% and 41.1%,respectively.ConclusionsCompared with the adjacent tissues,the expression level of hsa＿circ＿001710 in gastric cancer decreased significantly,which was significantly related to tumor size(p = 0.016)and tumor differentiation(p = 0.028).Accordingly,the down-regulation of hsa＿circ＿001710 expression may have significance in the growth and differentiation of gastric cancer lesion,and it has certain significance in the diagnosis and treatment of gastric cancer.