Expression And Clinical Significance Of IFIT2 In Cardia Adenocarcinoma

Background Gastric cardia adenocarcinoma as one of the digestive tract malignant tumor, its trend incidence is rising year by year. Because of the lack of effective early diagnosis index, the GCA metastasis had occurred for the most patients when seeing a doctor and they had poor clinical outcomes. Therefore, the further exploring the alterations of genes related to development and progression of GCA should be much accounted of. IFIT2 (IFN-induced protein with tetratricopeptide repeats 2) is a member of IFIT family genes,which has a large number of biological characteristics such as a protection against the viral infections, immune regulation, resist cell migration, and promote cell apoptosis. An increasing body of evidence indicates that IFIT2 have a critical role in protecting against viral infections; however, the role of IFIT2 in the development and progression of GCA remains largely unclear.Objective In this study, we examined IFIT2 expression in Gastric cardia adenocarcinoma tissue specimens and adjacent normal mucosa tissue via immunohistochemistry-, Western blotting and RT-PCR. in order to investigate the important role of IFIT2 in GCA in early diagnosis, clinical targeted treatment and prognosis.Methods Our specimens were randomly selected from 60 patients, who accepted Radical resection of cardiac carcinoma-in the first affiliated hospital of henan university of science and technology during the Feb 2012 to Oct 2013. There are 31 male and 29 female patients. The protein levels expressions of IFIT2 in GCA and normal esophageal mucosa tissue was examined by immunohistochemistry and western blotting assay. The gene levels expressions of IFIT2 were examined by qRT-PCR. The data were analysized by software SPSS 17.0, chi-square test or rank-sum test, P<0.05 was considered asstatistically significant value.Results 1. Immunohistochemistry:The positive expression rates of IFIT2 in 60 tissues paired non-cancerous tissues are 25% (15/60)、70% (42/60) respectively, The down-regulation of IFIT2 expression in tumor tissues was positively correlated with degree of differentiation (chi-square test P= 0.015). There was no significant correlation among the expression of IFIT2 with sex and Clinical staging (TNM) (chi-square test P > 0.015).2. Western blotting:Eight specimens in pairs were selected randomly from 60 GCA, in GCA tissue and adjacent normal mucosa tissue specimens. The study was done in order to confirm the differences expression of IFIT2 in GCA tissues and paired adjacent normal mucosa tissue specimens by Western blot. The experiment results from Western blot were firstly scanned by gel imaging system (721 BRO05922 type, the BIO-RAD company in Americen). then we measure a stripe image grey value from the expression of in GCA tissues and paired adjacent normal mucosa tissue specimens through the gel image analysis software. The protein levels expression of IFIT2 in two groups was correlated (rank-sum test.Z=2.474,P=0.018)3. qRT-PCR:Thirty specimens in pairs were selected randomly from 60 GCA tissues paired adjacent normal mucosa tissue specimens.The different expressions of IFIT2 in thirty specimens in pairs were amplified, analysed and validated through the application of real-time fluorescent quantitative PCR. by the the designed primers from primer 5 software. GAPDH was used as an internal control.the Ct values of IFIT2 expression were detected by qRT-PCR. The expression of IFIT2 in two groups was correlated (Student’s / test. t =3.954, <0.001).Conclusion 1. IFIT2 have a critical inhibition role in the development and progression of GCA.2. The down-regulation of IFIT2 expression may participate in the development of cardia cancer.3. The relation of IFIT2 expression with TNM staging and tumor progression needs to be further researched.