The CTP-NPRL2 Fusion Protein Inhibit Migration,Invasion Of Primary RCC Cells

Objective Effect of migration ? invasion after NPRL2(Nitrogen permease regulator 2-like)fusion protein into primary RCC cells via CTP(Cytoplasmic transduction peptide),infuence on EMT of RCC cells.Methods Primary RCC cells were cultured and identified from renal tumor tissue.Prokaryotic expression plasmids p ET15b-CTP-NPRL2 and p ET15b-NPRL2 were built and transfected into E.coli BL21,expressed CTP-NPRL2 and NPRL2 fusion protein.Fusion protein were tested by western blot.Primary RCC cells were divided into three group CTP-NPRL2?NPRL2?Blank control group treated with CTP-NPRL2?NPRL2 fusion protein solution and pbs respectively.Cell invasion and migration ability were detected by Transwell.The Raptor ?Vimentin?Fibronectin?E-cadherin were detected by real-time PCR and Western blot for the analysis of related m RNA and protein expression.Results Primary RCC cells were cultured successfully;The sequencingResult and Western Blot showed Prokaryotic expression plasmids were constructed succeefully;Green fluorescence appeared and located in cytoplasm in CTP-NPRL2 group rather than NPRL2 group or blank control group.The Transwell result showed the invasion and migration ability of Primary RCC cells could be significantly inhibited treated with CTP-NPRL2(P<0.05).the results showed CTP-NPRL2 fusion protein could decrease the expression of Raptor ? Vimentin and Fibronectin but increase the expression of E-cadherin in the level of m RNA and protein expression.Conclusion NPRL2 could regulated EMT of Primary RCC cells possiblely via Raptor depress the activity of m TORC1.