The Investigation Of Relationship Regulation Mechanism Between The Level Of PTEN/Survivin Expression In RA FLS

Object ives:To construct pc-PTEN expression vector and U6-sur interference vector and study the relevant regulation mechanism and the effects of PTEN or survivin in RA FLS, which provide experimental basis for the treatment of RA.Methods:Pc-PTEN expression vector and U6-sur interference vector were constructed. After RA FLS were isolated, RA FLS from passage three to seven were used. The gene or protein expression of PTEN or survivin of RA FLS were detected by RT-PCR or western bloting after transfected with pc-PTEN expression vector or U6-sur interference vector for24hour. CCK8assay and scratch test were performed to determine the proliferation and mortality of RA FLS after transfected, and the mRNA expression of MMP-1、MMP-3、MMP-9. IL-6were detected by RT-PCR.Result:1.The sequencing analyses demonstrated that both of the insertions sequencethe were perfectly corrected, pc-PTEN expression vector and U6-sur interference vector were successfully constructed.2.It showed a significant increasing of PTEN in both mRNA and protein levels after RA FLS was transfected with pc-PTEN expression vector for24hour, and a significant decrease of survivin in both mRNA and protein levels, and after transfected with U6-sur interference vector,the gene and protein expression of survivin was significantly decreased.3. Transfected with pc-PTEN expression vector or U6-sur interference vector can inhibite the proliferation and the migration of RA FLS, and the mRNA expression of MMP-1、MMP-3、MMP-9、IL-6of RA FLS showed a significant decrease.Conelusion:These results demonstrated that the pc-PTEN expression vector and U6-sur interference vector were successfully constructed;Both of the vectors can express in RA FLS;Transfected with pc-PTEN expression vector or U6-sur interference vector can inhibite the proliferation、migration、invasion and and inflammation of RA FLS. The PTEN/survivin cell-signalling pathway may exist in RA FLS.