Lü Xiaoying, Huang Yan, Qu Yayun, Zhang Yiwen, Zhang Zequn
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China.
SQ Medical Device Co., Ltd, Nanjing 210008, China.
Regen Biomater. 2020 Aug 31;7(6):553-565. doi: 10.1093/rb/rbaa030. eCollection 2020 Dec.
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.
本文的目的是整合转录组分析和蛋白质组学概况,揭示并比较PC12细胞在壳聚糖膜和胶原/壳聚糖膜表面生长的不同分子机制。首先,制备了壳聚糖膜和胶原/壳聚糖膜。随后,进行了细胞活力测定;在胶原/壳聚糖膜上培养24小时的PC12细胞的细胞活力显著高于在壳聚糖膜上培养的细胞。然后,通过cDNA微阵列分析,壳聚糖膜和胶原/壳聚糖膜表面PC12细胞的差异表达基因数量分别为13349个和5165个。接下来,生物学通路分析表明,差异表达基因涉及40条与细胞黏附和生长直接相关的通路。整合转录组分析和我们之前的蛋白质组学分析表明,细胞外基质-受体相互作用、黏着斑和肌动蛋白细胞骨架调节这三条生物学通路在蛋白质吸附、细胞黏附和生长过程中受到调控。材料表面吸附的蛋白质通过调节这三条通路中相关受体基因的表达,进一步影响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较强的抑制作用,导致其表面细胞活力显著降低;相反,胶原/壳聚糖膜更有利于促进PC12细胞的黏附和生长,从而使细胞活力更高。