The Study On LOXL2 Mediating Snail Protein SUMOylation Of The Cholangiocarcinoma Cell Line

Objective:(1)To establish the cell model subcell line of EX-LOXL2-QBC939 that stably expressed the protein,in order to find out the bionomics of LOXL2 and the relationship between LOXL2 and occurrence and development of CC.(2)To observe the impact of EX-LOXL2 to stability of Snail in QBC939 cells.(3)To observe the impact of EX-LOXL2 to stability of Snail in QBC939 cells.Methods:(1)To obtain the lowest concentration of G418 that kill all QBC939 cytes by using the puromycin dosage-reaction test.(2)To establish the cell model QBC939 that stably expressed LOXL2 by Lipofection.We obtained the positive clons by the G418 selection base on its lowest killing concentration.To culture the QBC939-control,QBC939-vector-LOXL2 and QBC939-EX-LOXL2.we detected the LOXL2 mRNA and protein by RT-PCR and western blot respectively,which will assure that we establish the QBC939-EX-LOXL2 cell model successfully.(3)By transfer transitly siRNA-SUM02/3 and siRNA-SUM02/3 vectors,we detected expression of LOXL2?SUM02/3 and Snail in QBC939-EX-LOXL2-control,QBC939-EX-LOXL2+siRNA-SUM02/3-vector and QBC939-EX-LOXL2+siRNA SUM02/3.(4)By transfer transitly EX-SENP1 and EX-SENP1 vectors,we detected expression of LOXL2?SENP1 and Snail in QBC939-EX-LOXL2-control,QBC939-EX-LOXL2+EX-SENP1-vector and QBC939-EX-LOXL2+EX-SENP1.Conclusion:(1)The stably cell model QBC939-EX-LOXL2 had been established successfully,which lay the foundation for us to study the bionomics of LOXL2 and the relationship between LOXL2 and occurrence and development of CC.(2)LOXL2 promote stability of Snail by inducing SUMOylation of Snail in CC cells.It provides a new target for prevention and treatment of invasion and metastasis of cholangiocarcinoma cells.