Expression Of UPF1 In Endometrial Carcinoma And Its Impact On MTOR Signaling Pathway

Backgroud Endometrial endometrioid carcinoma(EEC)is one of the most common malignant gynecological tumors.EECs can be divided into different subtypes and exhibit unique pathologies and different biological behaviors.However,UPF1 is a key protein factor in the nonsense-mediated m RNA degradation pathway(NMD)and its role in cancer remains to be studied.Therefore,this study hopes to study the pathogenesis,migration and invasion of UPF1 in endometrioid carcinoma,and can further understand the pathogenesis of endometrioid carcinoma,thus providing help for clinical treatment and diagnosis.Objective 1.To detect the RNA expression level of UPF1 in EEC tissues and adjacent tissues and different sources of EEC cell lines;2.Verify the effect of UPF1 on cell function;3.Subcutaneous tumorigenesis in nude mice verifies the effect of UPF1 on tumorigenicity;4.To investigate the effect of UPF1 on the mTOR signaling pathway.Methods 1.Firstly,qRT-PCR was used to detect the RNA expression level of UPF1 in human fresh EEC tissues,paracancerous tissues and EEC cell lines.2.Immunohistochemical method was used to detect the expression of UPF1 in EEC tissues and paracancerous tissues.3.UPF1 overexpression plasmid was constructed,and transient transfection promoted the expression of UPF1 in EEC cell line(RL952,ishikawa);si UPF1 was designed to inhibit the expression of UPF1 in EEC cell line(RL952,ilishkawa);4.Biological function test: After transient transfection of UPF1 over-expression plasmid and si UPF1 to inhibit UPF1 expression,the biological behavior of EEC cells was detected.(1)Flow cytometry was used to detect the cell cycle and apoptosis after overexpression of UPF1 and inhibition UPF1 expression by si UPF1.(2)Using CCK-8 to detect the change of EEC cell growth curve;(3)The effects of overexpression of UPF1 and inhibition of UPF1 expression on migration and invasion of EEC cells were detected by cell scratches and Transwell migration and invasion assays.5.Western blot was used to detect the expression of mTOR and downstream proteins(p70s6k/p-p70s6k)after changing the expression of UPF1 in EEC cell line.6.Construct a stable cell line to perform subcutaneous tumorigenesis in nude mice;7.Statistical analysisResults 1.qRT-PCR detection found that UPF1 RNA expression level in EEC tissues was higher than that in paracancerous tissues.2.Cellular function tests demonstrated that UPF1 can promote the proliferation and invasion of EEC cells,and inhibiting the expression of UPF1 can inhibit the proliferation,invasion and migration of EEC cells.3.Inhibition of UPF1 expression can inhibit the growth rate of subcutaneous tumor growth in nude mice.4.Western blot results showed that UPF1 can upregulate mTOR,activate mTOR/p70s6k/p-70s6 k signaling pathway,and activate mTOR to promote cell proliferation.Conclusions 1.UPF1 is highly expressed in EEC tissues compared with adjacent tissues;2.UPF1 as an oncogene in EEC can promote the proliferation,migration and invasion of EEC cells and inhibit the apoptosis rate of EEC cells.3.Inhibition of UPF1 expression in nude mice inhibits tumor growth;4.UPF1 can upregulate mTOR expression and activate the mTOR signaling pathway to promote cell proliferation.