Efficient inhibition of fibroblast proliferation and collagen expression by ERK2 siRNAs.
Transforming growth factor-beta1 and fibroblast growth factor-2 play very important roles in fibroblast proliferation and collagen expression. These processes lead to the formation of joint adhesions through the SMAD and MAPK pathways, in which ERK2 is supposed to be crucial. Based on these assumptions, lentivirus (LV)-mediated small interfering RNAs (siRNAs) targeting ERK2 were used to suppress the proliferation and collagen expression of rat joint adhesion tissue fibroblasts (RJATFs). Among four siRNAs examined, siRNA1 caused an 84% reduction in ERK2 expression (p<0.01) and was selected as the most efficient siRNA for use in this study. In subsequent experiments, significant downregulation of types I and III collagen were observed by quantitative RT-PCR and Western blot analyses. MTT assays and flow cytometry revealed marked inhibition of RJATF proliferation, but no apoptosis. In conclusion, LV-mediated ERK2 siRNAs may represent novel therapies or drug targets for preventing joint adhesion formation.
Li
Fengfeng
F
Department of Orthopaedics, The Sixth Affiliated People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China. fengmale@yahoo.com.cn
Fan
Cunyi
C
Cheng
Tao
T
Jiang
Chaoyin
C
Zeng
Bingfang
B
D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D049109:Cell Proliferation; D024042:Collagen Type I / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D024061:Collagen Type III / Q000096:biosynthesis; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism / Q000502:physiology*; D015316:Genetic Therapy; D006801:Humans; D007592:Joint Diseases / Q000628:therapy; D016086:Lentivirus; D019950:Mitogen-Activated Protein Kinase 1 / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000494:pharmacology
D024042:Collagen Type I; D024061:Collagen Type III; D034741:RNA, Small Interfering; D016212:Transforming Growth Factor beta; D016222:Fibroblast Growth Factor 2; D019950:Mitogen-Activated Protein Kinase 1