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Integrated transcriptomic and proteomic study on the different molecular mechanisms of PC12 cell growth on chitosan and collagen/chitosan films

机译:壳聚糖和胶原蛋白/壳聚糖薄膜PC12细胞生长不同分子机制的集成转录组和蛋白质组学研究

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摘要

The purpose of this article is to integrate the transcriptomic analysis and the proteomic profiles and to reveal and compare the different molecular mechanisms of PC12 cell growth on the surface of chitosan films and collagen/chitosan films. First, the chitosan films and the collagen/chitosan films were prepared. Subsequently, the cell viability assay was performed; the cell viability of the PC12 cells cultured on the collagen/chitosan films for 24 h was significantly higher than that on the chitosan films. Then, with cDNA microarray, the numbers of differentially expressed genes of PC12 cells on the surface of chitosan and collagen/chitosan films were 13349 and 5165, respectively. Next, the biological pathway analysis indicated that the differentially expressed genes were involved in 40 pathways directly related to cell adhesion and growth. The integrated transcriptomic and our previous proteomic analysis revealed that three biological pathways—extracellular matrix–receptor interaction, focal adhesion and regulation of actin cytoskeleton—were regulated in the processes of protein adsorption, cell adhesion and growth. The adsorbed proteins on the material surfaces further influenced the expression of important downstream genes by regulating the expression of related receptor genes in these three pathways. In comparison, chitosan films had a strong inhibitory effect on PC12 cell adhesion and growth, resulting in the significantly lower cell viability on its surface; on the contrary, collagen/chitosan films were more conducive to promoting PC12 cell adhesion and growth, resulting in higher cell viability.
机译:本文的目的是整合转录组分析和蛋白质组学曲线,并揭示并比较壳聚糖膜和胶原/壳聚糖膜表面上PC12细胞生长的不同分子机制。首先,制备壳聚糖膜和胶原/壳聚糖膜。随后,进行细胞活力测定;在胶原/壳聚糖膜上培养的PC12细胞24小时的细胞活力显着高于壳聚糖膜上的细胞。然后,通过cDNA微阵列,壳聚糖表面和胶原/壳聚糖膜表面上的PC12细胞的差异表达基因的数量分别为13349和5165。接下来,生物途径分析表明,差异表达的基因涉及40种与细胞粘附和生长有关的途径。综合转录组和我们之前的蛋白质组学分析表明,在蛋白质吸附,细胞粘附和生长过程中调节三种生物途径 - 细胞外基质受体相互作用,局灶性粘附和调节肌动蛋白细胞骨架 - 受到调节。材料表面上的吸附蛋白进一步影响了重要下游基因的表达,通过调节在这三种途径中的相关受体基因的表达。相比之下,壳聚糖薄膜对PC12细胞粘附和生长具有很强的抑制作用,导致其表面上显着降低的细胞活力;相反,胶原蛋白/壳聚糖膜更有利于促进PC12细胞粘附和生长,导致更高的细胞活力。

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