RESUMO
Objective To discuss the relationship between the expression of PLOD2 and the formation of collagen I in the scar. Methods To detect the mRNA expression of PLOD2 and COLIα1 in the scar and normal skin tis-sue, as well as the content of COLIα1 by RT-PCR and Western blot; design and synthesize three kinds of PL0D2 ASODN targeted at the translational initiation, the joint of the first and the second exons and termination codon re-spectively , transfected into NIH/3T3 mouse fibroblast cell and detect the mRNA expression of COLIα1 and forma-tion of collagen I by RT-PCR and Western blot. Results The expression of PLOD2 in the scar tissue was signifi-cantly higher than that in the normal tissue ( P < 0. 05 ) , three kinds of ASODNs showed inhibitory effects on the mRNA expression of COLIα1 and formation of collagen I in fibroblast (P <0. 05) , during which the repressive ef-ficiency of ASODN targeted against the translational initiation was the highest (P <0. 001) ; while the mRNA ex-pression level of COLIα1 did not change in the scar tissue, normal tissue and the groups treated with three kinds of ASODN. Conclusion The high expression of PLOD2 is tightly related to the formation of collagen I , PLOD2 ASODN can suppress the formation of collagen I post-transcriptionally.
RESUMO
Objective To investigate the role of wild-type XPD in SMMC-7721 hepatoma cells,its re-lationship with wild-type p53.CDK7 and c-myc.And the apoptosis mechanism of SMMC-7721 hepatoma cells.Methotis We inserted the human length XPD into the pEGFP-N2 plasmid vector which expresses the green fluorescence protein(GFP).And the pEGFP-N2 and pEGFP-N2-XPD were transfected into SMMC-7721 hepa-toma cell lines stably.Cel lines for omparison were matched on the sanle genetic background and passage.The expression of wild-type XPD,CDK7,p53,c-myc waft detected by RT-PCR,Westem blot.Cell growth wag detet-ed by MTT FCM wag employed for examining the cell cycle and apoptosis of the transfected SMMC-7721 hepa-toms cells.Results ①SMMC-7721-pEGFP-N2-xPD and SMMC-7721-pEGFP-N2 express the green fluores-cence protein which could be detected under the immunoffuorescence microscope.②RT-PCR The relative ex-\pression of p53 mRNA,XPD mRNA in the SMMC-7721,SMMC-7721-pEGFP-N2 and SMMC-7721-pEGFP-N2-XPD,the relative expression of p53 mRNA,XPD mRNA in the SMMC-7721-pEGFP-N2-XPD was signifiantly higher than other two oontrols(P<0.01).Othewise,the relative expression of CDK7 mRNA,c-myc mRNA Wag signifi-antlv lower than other two oontrols(P<0.01),the difference of two controls was not signifiant(P>0.05). ③Western blot The relative expression of p53,XPD protein was signifiantly higher than other two oontrols (P<0.01).Othewise,the relative expression of CDK7,c-Myc protein was signifiantly lower than other two oontrols(P<O.01),the difference of two controls Wag not signifiant(P>0.05).④MTT The ressuh showedthat the proliferative ability of SMMC-7721-pEGFP-N2-XPD was much more descreaged than other two oontrols (P<0.05),the other two oontrols were not different(P>0.05).⑤FCM Wild-type XPD gene could change the cell8 and induce apoptitise in vitro though the result of FCM.Conclusions The pEGFP-N2 and pEGFP-N2-XPD were transfected into SMMC-7721hepatoma cell lines stably,wild-type XPD could decreaSe the expres-sion of CDK7,c-myc and increase the expression of p53.Wild-type.XPD could inhibit the activity of cell growth and change cell cycle,as well as induce cell apoptosis in SMMC-7721 hepatoma cell line in vitro.