Effects Of Wnt4 On FOXN1 And The Effect Of Down-regulation On Cell Apoptosis In Thymoma

Objective:Wnt signaling pathway regulates the development of thymic epithelial cells by modulating the expression of FOXN1.Thymoma is the most common anterior mediastinal tumor,more types of histology,and all kinds of subtypes can occur invasive growth,and a small number of them may transter distant,although its has longer course of disease,the effect is relatively good,but It still a malignant tumor,its complication such as myasthenia gravis and other autoimmune diseases has great harm to human,affecting the patients quality of life seriously.Wnt4 and FOXN1 are two factors that have been closely related to the growth of thymoma in recent years.In order to determine whether Wnt4 and FOXN1 are involved in the pathogenesis of thymoma.In this study,the expression of Wnt4 and its access factor in human thymoma cells was down-regulated by si RNA,Which are synthesis by chemical method.The m RNA and protein expression of FOXN1 in thymoma cells was detected by PCR and Western-Blot method,thus,we can identify the signal pathway which wnt4 regulate thymoma growth,and to investigate the effect of Wnt4 and FOXN1 on the apoptosis of thymoma cells by flow cytometry.Methods:1?According c DNA of Wnt4?JNK ?Tcf-4 and FOXN1 in Gene Banks,constructed and synthesized Wnt4-si RNA?JNK-si RNA?Tcf-4-si RNA and FOXN1-si RNA,For each gene,3 pairs of si RNA were synthesized and cultured in vitro with thymoma cells with Lipofectamine TM 2000 and transfected with si RNA targeting Wnt4,JNK,Tcf-4 and FOXN1,The expression of Wnt4,JNK and Tcf-4was detected by PCR,The follow-up experiments were carried out to filter the best interference si RNA effect of Wnt4,JNK,Tcf-4 and FOXN1,detecting the expression level of FOXN1 m RNA and proterin after interference each group of genes by RTq PCR and Western-bloting.2?Wnt4-si RNA and FOXN1-si RNA were transfected into human thymoma cells in vitro,Flow cytometry was used to detect the effect of Wnt4 gene and FOXN1 expression on the apoptosis of thymoma cells Result:1?Compared with the control group,the best interence of Wnt4-si RNA,JNKsi RNA,Tcf-4-si RNA and FOXN1-si RNA is Wnt4-986,JNK-722,Tcf-4-1717 and FOXN1-1069,respectively.The inhibition rates were 56.7%,72.6%,63.2%and52.3%,respectively,after deregulation Wnt4 and JNK gene,the expression of FOXN1 m RNA and protein in human thymoma cells was notablely lower than that in the blank group(P<0.05),and after down-regulation Tcf-4 gene,FOXN1 m RNA and protein expression were not different from the control group(P>0.05).2?Compared with the control group,the number of apoptotic cells in human thymoma cells was significantly increased after interference with Wnt4 and FOXN1 gene,the difference was statistically significant(P <0.05)Conclusion:1?Wnt4-986,JNK-722,Tcf-4-1717 and FOXN1-1069 were able to inhibit the expression of Wnt4,JNK,Tcf-4 and FOXN1,respectively.The expression of FOXN1 in human thymoma cell line was significantly decreased after interfering with Wnt4 and JNK genes by si RNA,suggesting that Wnt4 and JNK could regulate the downstream FOXN1 in thymoma cell lines,also suggesting that the expression of transcription factor FOXN1 in thymoma cells cultured in vitro may be achieved by nonclassical signal pathway—Wnt / JNK pathway.2?Interfering with Wnt4 gene and transcription factor FOXN1 in human thymoma cell lines can promote thymoma cell apoptosis,suggesting that change of Wnt4 gene and transcription factor FOXN1 expression may be related to the occurrence and development of thymoma,suggesting that Wnt4 gene may be plays a role in thymoma through transcription factors FOXN1.