Versican targeting by RNA interference suppresses aggregative growth of dermal papilla cells

维斯坎 RNA干扰 细胞生物学 细胞外基质 化学 蛋白多糖 硫酸软骨蛋白多糖 分子生物学 核糖核酸 生物 生物化学 基因
作者
Ming Feng,Guihong Yang,Jinjin Wu
出处
期刊:Clinical and Experimental Dermatology [Oxford University Press]
卷期号:36 (1): 77-84 被引量:19
标识
DOI:10.1111/j.1365-2230.2010.03917.x
摘要

Summary Background. Dermal papilla cells (DPCs) are specialized fibroblasts found in the hair follicle papilla, which are associated with the development and cycle regulation of hair follicles (HFs). DPCs exhibit a multilayer aggregative growth character, which is closely related to induction of HF formation. Versican, a large chondroitin sulphate proteoglycan and one of the major components of the extracellular matrix, is involved in the formation of HF. Methods. To confirm the relationship between versican and the aggregative growth of DPCs, we first induced and established an aggregative cell model in DPCs in vitro, with cells taken to passage 8. Simultaneously, aggregative passage 2 DPCs and nonaggregative passage 8 DPCs were selected as parallel controls. RNA interference (RNAi) targeted to versican was used in passage 2 DPCs using a lentiviral vector. Reverse transcriptase (RT)-PCR and western blotting were used to assay the expression of versican in DPCs. Results. RNAi targeted to versican efficiently suppressed the aggregative growth of passage 2 DPCs, and the inhibitory effect was significant 3 days after RNAi treatment. The mRNA and protein levels of versican were also downregulated in passage 2 DPCs, and were lower than levels in nonaggregative passage 8 DPCs. Notably, the aggregative growth of nonaggregative passage 8 DPCs was restored after induction in a 1 : 1 v/v mixture of fresh DMEM and medium recycled from a previous passage. Conclusion. Versican is a key gene for the aggregative growth of DPCs, and might be significant in the regeneration of HF.
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