The Role And Mechanism Of UPAR In Tumor Multidrug Resistance

Objective:Cancer is a huge disease problem faced by human beings.Chemotherapy is an effective treatment method for cancer and is widely used.However,tumor cells can easily develop resistance to chemotherapeutic drugs,especially the occurrence of multidrug resistance,which can easily lead to failure in chemotherapy.uPAR plays an important role in the occurrence and development of tumors,but there are few reports on the relationship between uPAR and tumor multidrug resistance.Therefore,this paper wants to study the role and mechanism of uPAR in multidrug resistance of tumors.Methods:1.We use R language programming analyse the RNA expression levels data and clinical prognosis data of uPAR in TCGA database.2.We use CRISPR-Cas9 technique knock out uPAR in HCT8/T and KB_V200. and we use the Gateway cloning technique construct a over-expressing vector of uPAR to overexpress uPAR in AGS and H1299.3.We use the methods of Scratch,Transwell,Softagar,MTT,phalloidin and DAPI double staining to detect the effect of changing uPAR on the ability of multidrug resistance,proliferation,migration,invasion in tumor cells.4.We use western blot technique to explore the mechanism of uPAR in tumor multidrug resistance.Results:1.TCGA data show that uPAR protein is highly expressed in tumor tissues,and the high expression of uPAR is associated with poor prognosis in lung cancer and kidney cancer patients.2.After knocking out uPAR in tumor multidrug resistant cell lines HCT8/T and B_V200.,the ability of multidrug resistance in tumor cells is weakened.However,after overexpression of uPAR in tumor cell lines AGS and H1299with low expression of uPAR,the ability of multidrug resistance in tumor cells is enhanced.3.After knocking out uPAR in tumor multidrug resistant cell lines HCT8/T and B_V200.,the tumor’s proliferation,migration,and invasion were weakened.However,when uPAR was over-expressed in tumor cell lines AGS and H1299with low expression of uPAR,the proliferation,migration and invasion of tumors were enhanced.4.After knocking out uPAR in tumor multidrug resistant cell lines HCT8/T and B_V200.the expression of EGFR,HER3,β-Catenin,p-ERK1/2（Thr202/Tyr204）,p-JNK（Thr183/Tyr185）,p-PTEN（Ser380）,BCL-XL,Snail will decreased.However,when uPAR was over-expressed in tumor cell lines AGS and H1299with low expression of uPAR,the expression of these protein will increased.Conclusion:TCGA data show that uPAR protein is highly expressed in tumor tissues,and he high expression of uPAR is associated with poor prognosis in lung cancer and kidney cancer patients.u PAR can promote the occurrence of multidrug resistance and the ability of proliferation,migration,invasion in tumors.what’s more,uPAR may affect directly or indirectly in the expression of epidermal growth factor receptor（EGFR）family;Wnt/β-Catenin,MAPK pathways;the phosphorylation level of PTEN tumor suppressor;anti-apoptotic pathways;epithelial-mesenchymal transition pathways to promote the occurrence of multidrug resistance and the proliferation,migration,invasion in tumors.