Establishment of a model of murine odontoblasts underexpressing PKD1 using shRNA.

B Thivichon-Prince, N Labert, Ml Couble, F Bleicher
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Abstract

We have previously shown that PKD1, the gene encoding Polycystin-1 (or TRPP1) is expressed in human odontoblasts and that this protein is localized at the primary cilium of the cell. Nevertheless, its function remain unclear in this cell even if studies on osteoblasts, osteocytes and chondrocytes give TRPP1 as a promising candidate for mechanotransduction in response to mechanical stress. Consequently, to evaluate the role of TRPP1 in this transduction process, we needed first to generate an in vitro murine model down expressing Pkd1. Using lentivirus-mediated shRNA technology, we obtained a 60% suppression of Pkd1 mRNA expression in transfected MO6-G3 cells associated with a decrease of cell proliferation. Thus, establishment of this murine odontoblast model underexpressing Pkd1 associated with applied mechanical forces (compression or shear stress) will allow us to go further in the determination of TRPP1 involvement in odontoblasts mechanotransduction.

利用shRNA建立低表达PKD1的小鼠成牙细胞模型。
我们之前已经证明,编码多囊蛋白-1(或TRPP1)的基因PKD1在人成牙细胞中表达,并且该蛋白定位于细胞的初级纤毛。然而,尽管对成骨细胞、骨细胞和软骨细胞的研究表明TRPP1作为机械应力响应的机械转导的有希望的候选者,其在该细胞中的功能仍不清楚。因此,为了评估TRPP1在这一转导过程中的作用,我们首先需要建立一个体外低表达Pkd1的小鼠模型。利用慢病毒介导的shRNA技术,我们在转染的MO6-G3细胞中获得了60%的Pkd1 mRNA表达抑制,并伴有细胞增殖降低。因此,建立与机械力(压缩或剪切应力)相关的低表达Pkd1的小鼠成牙细胞模型将使我们能够进一步确定TRPP1参与成牙细胞机械转导。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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