| Background:Ovarian cancer is the highest mortality gynecological malignant tumor,and it is also a highly metastatic disease.Transforming growth factor-β1(TGF-β1)is the main driving force of epithelial-mesenchymal transformation,promoting tumor cell metastasis and drug resistance.Tumor necrosis factor inducible protein 8(TNFAIP8)protein like 2(TIPE2)is not only a negative regulator of innate and acquired immunity,but also involved in the occurrence and development of different tumors.However,the role of TIPE2 in ovarian cancer and whether TIPE2 reverses the epithelial-mesenchymal transformation induced by TGF-β1 in ovarian cancer cells are still unclear.This study aims to explore the role of TIPE2 in ovarian cancer and to find potential targets for the diagnosis and treatment of ovarian cancer.Methods:(1)Immunohistochemical method(IHC)was used to detect the expression of TIPE2 in control group(normal fallopian tube tissue and normal ovarian tissue)and experimental group(serous and mucinous ovarian cancer tissue).The relationship between the expression of TIPE2 and pathological type,as well as patient’s age,tumor size,tumor differentiation degree,FIGO stage,disease-free survival rate and overall survival rate of ovarian cancer was analyzed.(2)The effects of TIPE2 overexpression/knockdown on the proliferation,migration and invasion of ovarian cancer cells were detected by EdU,clone formation,Transwell migration and invasion assays.(3)Western-blot was used to detect the effect of TIPE2 overexpression/knockdown on the expression of EMT-related markers and transcription factors.(4)Western-blot was used to detect the effect of TIPE2 overexpression/knockdown on the expression of proteins related to TGF-β1 expression.(5)Rescue experiment was used to detect whether TIPE2 played a role via Smad2-EMT-related pathway,and co-IP method was used to verify the combination of TIPE2 and Smad2.(6)Peritoneal metastatic xenograft model of nude mice was used to detect whether the expression of TIPE2 inhibited the metastasis of ovarian cancer cells in vivo.(7)Transwell migration and invasion assay was used to detect the effect of TGF-β1 on migration and invasion of ovarian cancer cells.Western-blot was used to detect the effect of TGF-β1 on EMT genesis and the expression of TGF-β1 pathway related proteins.(8)Transwell migration and invasion assays were used to detect whether TIPE2 overexpression could reverse TGF-β1-induced migration and invasion of ovarian cancer cells.Western-blot was used to detect whether TIPE2 overexpression could reverse TGF-β1-induced EMT occurrence and the expression of TGF-β1 pathway-related proteins in ovarian cancer cells.(9)The correlation between TIPE2 and TGF-β1 was detected by qRT-PCR,Western-blot,ELISA and IHC of mouse tumor.Results:(1)The expression of TIPE2 was decreased in ovarian cancer tissues,and its expression was closely related to pathological type,patients’ age,tumor differentiation degree and FIGO stage,but not related to tumor size,disease-free survival rate and overall survival rate.(2)TIPE2 had no significant effect on the proliferation and colony formation ability of ovarian cancer cells.(3)TIPE2 inhibited the migration and invasion of ovarian cancer cells.(4)TIPE2 upregulated the expression of epithelial cell marker,and downregulated the expression of mesenchymal cell markers and EMT-related transcriptional factors.(5)TIPE2 inhibited Smad2-EMT-related pathway by combining with Smad2.(6)TIPE2 inhibited the metastasis of ovarian cancer cells in the peritoneal metastatic xenograft model of nude mice.(7)TIPE2 promoted the migration,invasion,EMT occurrence and the expression of TGF-β1 pathwayrelated proteins in ovarian cancer cells.(8)TIPE2 reversed TGF-β1-induced migration,invasion,EMT occurrence and the expression of TGF-β1 pathway-related proteins in ovarian cancer cells.(9)There was a negative correlation between TIPE2 and TGF-β1.Conclusion:TIPE2 plays an important role in the metastasis of ovarian cancer.TIPE2 can inhibit the epithelial-mesenchymal transformation induced by TGF-β1 by targeting Smad2EMT pathway,thus inhibiting the migration and invasion of ovarian cancer cells.Innovation and significance:The expression of TIPE2 in ovarian cancer is decreased.We first found that TIPE2 not only inhibits EMT,migration and invasion of ovarian cancer by combining with Smad2,but also reverses EMT,migration and invasion induced by TGF-β1.In addition,we first found that there is a negative regulation between TIPE2 and TGF-β1,which provides a new and effective target for the diagnosis and treatment of ovarian cancer. |
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