TIPE2 Mediates Proliferation? Migration?invasion And Apoptosis Of Human Rectal Adenocarcinoma Cells

BackgroundRectal cancer is the most common malignant gastrointestinal tumor.The incidence of the disease has risen rapidly around the world in recent years.In China,the median age of rectal cancer onset is around 45 years old,and the incidence of young people has an increasing trend.The traditional surgical therapy about rectal cancer has its drawbacks,and the research on rectal cancer still has problems such as unclear mechanism of occurrence,unknown treatment target,lack of surface markers,so we turn our eyes to the target treatment of rectal cancer and devote ourselves to finding relevant targets.Adenocarcinoma is the most common type of rectal cancer,so we uesd the most commonly human rectal adenocarcinoma cell lines HR8348 and SW837 in this study.The TNFAIP8(tumor necrosis factor-a-induced protein-8 family,also known as the TIPE family,is a newly reported immunomodulatory and tumor regulator..TIPE2(TNF-?-induced protein 8-like 2)is a member of the TIPE family and is considered to be a new negative regulator of immune and inflammatory homeostasis,which is closely related to the development of tumor and may be a new molecular target for tumor therapy.The results of the existing studies show that the expression of TIPE2 in different tumors is different,but it has inhibitory effect.At present,there is no research related to rectal cancer at home and abroad,in order to understand the role of TIPE2 in the development of rectal cancer,this study discussed the expression of TIPE2 in human rectal adenocarcinoma and its effect on the proliferation and apoptosis of human rectal adenocarcinoma cells.PurposeThe expression of TIPE2 in human rectal adenocarcinoma and its effects on the proliferation,migration,invasion and apoptosis of human rectal adenocarcinoma were studied,and the mechanisms for causing change were discussed.MethodsThe expression of TIPE2 in human rectal adenocarcinoma was detected by immunohistochemistry and Western blot.The overexpression of TIPE2,knockdown and the corresponding blank control plasmid were transferred into human rectal adenocarcinoma cell line HR8348 and SW837 cells,respectively.Then stable cell lines were obtained by screening with G418.The effect of TIPE2 on cell proliferation was detected by MTS and EDU.The influence on the ability of cell cloning was tested by plate cloning experiment.Through scratch,transwell,soft agar colony formation experiment,invasion test,the influence of invasion on cell migration and invasion ability is detected.Tunel staining was used to detect the effect on apoptosis.The expression of apoptosis,autophagy related proteins and Wnt/?-Catenin?TGF-?/Smad2/3pathway were detected by western blot.The effect of TIPE2 on the proliferation of transplanted tumor in nude mice was detected by tumor formation assay.Results1.Immunohistochemical and Western blot results showed that the expression of TIPE2 in cancer tissues was higher than that in adjacent cancer tissues.2.MTS and EDU results showed that overexpression of TIPE2 could inhibit cell proliferation and silencing could promote cell proliferation.3.Scratch and transwell results showed that overexpression of TIPE2 could weaken cell migration and silencing could enhance cell migration.4.Soft agar colony formation and invasion results showed that over-expression of TIPE2 could weaken invasion ability,and invasion ability could be enhanced by silencing.5.Tunel staining showed that overexpression of TIPE2 promoted apoptosis,while silencing inhibited apoptosis.6.Western blotting results showed that the expression of Cleaved Caspase3 and Cleaved PARP increased after TIPE2 overexpression,while the expression of Wnt3 a decreased,and the phosphorylation levels of ?-Catenin and Gsk-3?decreased.Therefore,overexpression of TIPE2 promoted apoptosis and may regulate apoptosis through the Wnt/?-Catenin signaling pathway.7.Western blotting results showed that the expression of LC3A/B,Beclin-1 decreased,the expression of p62 increased,the phosphorylation level of Smad2 and Smad3 decreased,and the expression of TGF-? decreased after overexpression of TIPE2.Therefore,overexpression of TIPE2 inhibited autophagy and may regulate autophagy through the TGF-?/ smad2/3 signaling pathway.8.The tumor volume and weight of nude mice in the TIPE2 overexpression group were smaller than those in the control group,and the positive rates of KI67 and CD31 were lower than those in the control group.ConclusionTIPE2 inhibits proliferation,migration and invasion of human rectal adenocarcinoma cells,and may regulate apoptosis and autophagy through Wnt/ ?-Catenin and TGF-? / Smad2/3 signaling pathways,thereby inhibiting the occurrence and development of human rectal adenocarcinoma.