Influence and mechanism of mechanical properties of extracellular matrix on biological behaviors of oral squamous carcinoma cells
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摘要:
目的：初步探索细胞外基质力学表征对口腔鳞癌细胞生物学行为的影响。方法：在不同力学性能的水凝胶上接种两种口腔鳞癌细胞CAL27和HSC3，通过光学显微镜观察细胞形态变化；利用免疫荧光观察层黏连蛋白β2（laminin β2，LAMB2）、Ⅵ型胶原α1（collagen type Ⅵ α1，COL6A1）以及E-钙黏蛋白（E-cadherin，CDH1）的表达定位以及表达水平；使用qPCR检测细胞多能性指标（SOX2、NANOG）、LAMB2、COL6A1、CDH1、整合素α6β4的亚基α6（integrin α6，ITGA6）和亚基β4（integrin β4，ITGB4）基因的表达水平。结果：不同力学性能的水凝胶会改变口腔鳞癌细胞的黏附性，高力学性能的水凝胶会增加细胞黏附效率。使用高力学性能水凝胶进行细胞培养时，细胞的生物学行为和细胞培养板培养的表型相似。在低力学性能水凝胶的作用下，CAL27细胞会形成紧密肿瘤细胞球，而HSC3细胞则会形成不紧密的细胞团块。使用低力学性能水凝胶进行细胞培养时，两种细胞中LAMB2和COL6A1都会高表达，并且LAMB2在细胞上的受体整合素α6β4表达也会增高。细胞多能性指标（SOX2、NANOG）和CDH1在CAL27细胞中会随水凝胶力学性能增加而降低，而HSC3细胞则相反。结论：细胞外基质的力学性能可以调节细胞黏附效率、细胞外基质合成、肿瘤细胞多能性和细胞间黏合等多种生物学行为。细胞外基质的力学性能可能是通过改变细胞外基质蛋白的表达来改变细胞的生物学行为，细胞生物学行为改变的差异还与细胞系有关。
Abstract:
Objective：This study aims to explore the influence and mechanism of mechanical properties of extracellular matrix on biological behaviors of oral squamous carcinoma cells. Methods：Two kinds of oral squamous carcinoma cells，CAL27 and HSC3，were cultured on hydrogel with different mechanical properties. Cell morphological changes were observed by microscope. The location and expression of laminin β2（LAMB2），collagen VI α1（COL6A1） and E-cadherin（CDH1） were detected by immunofluorescence. The mRNA expression level of SOX2，NANOG，LAMB2，COL6A1，integrin α6β4（ITGA6 and ITGB4） and CDH1 were detected by qPCR. Results：The mechanical properties of hydrogel changed the adhesion of oral squamous carcinoma cells，and the high mechanical properties of hydrogel increased cell adhesion. The biological behavior of cells on hydrogel with high mechanical properties is similar to those on traditional cell culture plates. CAL27 formed tight multicellular tumor spheroid on hydrogel with low mechanical properties，while HSC3 formed loose cell clusters. On hydrogels with low mechanical properties，the expression of LAMB2，COL6A1，integrin α6β4 in the two kinds of cells were increase. Cell stemness markers（SOX2，NANOG）and E-cadherin（CDH1）in CAL27 decreased with mechanical properties increased in hydrogels，whereas HSC3 were opposite. Conclusion：The mechanical properties of extracellular matrix can regulate cell adhesion，extracellular matrix protein synthesis，tumor cell stemness and intercellular adhesion. The mechanical properties of extracellular matrix may change the biological behavior of cells by changing the protein expression of extracellular matrix. However，the specific cell biological behavior is related to the specificity of cell line.
