Expression Of FBXW 11 In Triple-negative Breast Cancer And Its Effect On Its Proliferation And Migration

Purpose Breast cancer is one of the most common malignant tumors,which has a significant impact on the quality of life and overall survival of women.Among them,the triple negative breast cancer,which is called triple negative breast cancer with negative expression of ER,PR and HER2,has the highest malignant degree,and its high rate of recurrence and metastasis is the main reason for the increase of mortality.F-frame / WD-40 domain protein 11 gene(FBXW11)is a member of the F-box protein family,which is characterized by about 40 amino acid motifs.FBXW11 plays a key role in cancer development by controlling the turnover of key proteins related to tumor cell proliferation and migration.Studies have shown that FBXW11 is very important for embryonic development and plays a key role in signal pathways such asβ-catenin/Wnt and NF-κ B.it also regulates HH and RAS signal pathways.FBXW11 also plays a role in a variety of tumors,such as skin tumors,cervical cancer,lung cancer,pancreatic cancer and so on.FBXW11 is closely related toβ-catenin/Wnt and NF-κ B signal pathways.However,the role of FBXW11 in breast cancer has not been reported.In this study,we investigated the expression of FBXW11 in triple negative breast cancer cell lines MDA-MB-231 and HCC-1937 and the effect of protein expression on cell proliferation and migration,which further proved that FBXW11 plays a role in triple negative breast cancer,so as to provide some theoretical reference for the diagnosis and treatment of breast cancer.Methods:1.The relative expression of FBXW11 in various cancer tissues was analyzed by bioinformatics method in The Cancer Genome Atlas(TCGA)database.Normal breast tissue samples and triple negative breast cancer tissue samples were extracted from TCGA database to analyze the expression of FBXW11 in clinical triple negative breast cancer samples and normal tissues.2.The full-length open reading frame gene of FBXW11 was amplified from breast cancer cell MDA-MB-231 by reverse transcription-polymerase chain reaction(RT-PCR),and then ligated and transformed to construct the recombinant GV141-FBXW11 vector for overexpression of FBXW11.The recombinant vector was confirmed by sequencing.3.Two kinds of FBXW11 overexpression cell lines and FBXW11 knockdown cell lines were constructed by lentivirus infection,and the overexpression and knockdown efficiency of FBXW11 were verified by real-time fluorescence quantitative PCR(RT-PCR)and Western Blot.4.Plate clone formation assay and MTT assay were used to detect the effect of FBXW11 protein expression on the proliferation of breast cancer cells.Scratch test was used to detect the effect of FBXW11 protein expression on the migration of breast cancer cells.Then Transwell chamber test was used to detect the effect of FBXW11 protein expression on invasion.5.Western blotting(Western Blot)was used to detect the difference ofβ-catenin and c-myc expression after the change of FBXW11 protein expression.6.The co-localization of FBXW11 and β-catenin was detected by immunofluorescence(Immunofluorescencetechnique,IF).Results1.Through the analysis of TCGA and HPA database,the m RNA level of FBXW11 in breast cancer tissue is lower than that in normal tissue and low expression in breast cancer tissue.2.The sequencing results showed that the recombinant vector was constructed successfully.3.RT-q PCR and Western Blot assay showed that FBXW11 was significantly overexpressed at m RNA level and protein level after lentivirus infection in two cell lines.After small interference transfection,Western Blot assay showed that the expression of FBXW11 protein decreased significantly.4.The MTT results of proliferation test showed that the growth rate of the cell line with overexpression of FBXW11 was significantly slower,while that of the cell line with FBXW11 knockdown was faster.5.Plate clone formation assay showed that the clone formation rate of FBXW11 overexpression cell line was lower than that of the control group,while the clone formation rate of FBXW11 knockout cell line was higher.6.Scratch test and Transwell chamber test showed that the overexpression of FBXW11 inhibited the migration and invasion of breast cancer cells,while the migration ability was enhanced after knocking down FBXW11.7.Immunofluorescence experiment showed that there was co-localization between FBXW11 and β-catenin.Western Blot showed that altered FBXW11 protein levels could affect the Wnt/β-catenin pathway.Conclusion1.The expression of FBXW11 protein in triple negative breast cancer cells and tissues is significantly lower than that in normal cells and normal tissues.2.FBXW11 overexpression cell line was obtained by screening after virus infection,and FBXW11 low expression cell line was obtained after small interference transfection.3.High expression of FBXW11 can inhibit the proliferation,migration and invasion of breast cancer cells,while low expression of FBXW11 can promote the proliferation,migration and invasion of breast cancer cells.4.FBXW11 Through the development of Wnt/β-catenin signaling pathway.5.FBXW11 Inhibition of tumor growth in nude mice...