Abstract

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.

中文翻译：

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壳聚糖和胶原/壳聚糖膜上PC12细胞生长的不同分子机制的转录组和蛋白质组学研究

摘要

本文的目的是整合转录组学分析和蛋白质组学概况，并揭示和比较壳聚糖膜和胶原蛋白/壳聚糖膜表面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，进行细胞生存力测定。胶原/壳聚糖膜上培养24 h的PC12细胞的细胞活力明显高于壳聚糖膜。然后，利用cDNA微阵列，壳聚糖和胶原/壳聚糖膜表面上的PC12细胞的差异表达基因数分别为13349和5165。接下来，生物学途径分析表明差异表达的基因参与了与细胞粘附和生长直接相关的40条途径。综合的转录组学和我们先前的蛋白质组学分析显示，在蛋白质吸附，细胞粘附和生长过程中，三个生物学途径（细胞外基质-受体相互作用，粘着斑和肌动蛋白细胞骨架的调节）受到调节。通过调节这三种途径中相关受体基因的表达，材料表面吸附的蛋白质进一步影响了重要下游基因的表达。相比之下，壳聚糖膜对PC12细胞的黏附和生长具有很强的抑制作用，从而导致其表面上的细胞活力大大降低。相反，胶原蛋白/壳聚糖膜更有利于促进PC12细胞的黏附和生长，从而提高细胞活力。