Abstract:

Zhang Jie*, Cao Yixin, Qin Jing, Wang Jianli, Chen Li. *Department of Dermatovenereologoy, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu, China Corresponding authors: Wang Jianli, Email: bl1@ntu.edu.cn; Chen Li, Email: guilan@ntu.edu.cn 【Abstract】 Objective To evaluate the effect of trypsin on the proliferation, migration and adhesion of a skin squamous cell carcinoma cell line A431. Methods Cultured A431 cells were divided into several experimental groups treated with trypsin at concentrations of 0.1, 1, 10 and 100 nmol/L for 24, 48 and 72 hours respectively, and a control group treated with DMEM complete medium only. Cell counting kit-8 （CCK8） assay was conducted to evaluate cellular proliferative activity to select the optimal concentration of trypsin. Then, some A431 cells treated with trypsin at the selected concentration for 24, 48 and 72 hours respectively （or 48 hours only） served as the experimental groups （or group）, and other A431 cells treated with DMEM complete medium served as the control group. Flow cytometry was performed to assess cell cycle distribution and proliferation index, fibronectin-based adhesion assay to estimate cell adhesive capacity, and wound healing assay and Transwell assay were conducted to evaluate the migratory capacity of cells in two- and three-dimensional space. Statistical analysis was carried out by using analysis of variance, paired samples t test and chi-square test. Results The proliferative activity of A431 cells increased along with the increase of trypsin concentrations, with the strongest increasing effect observed at 100 nmol/L. After treatment with 100 nmol/L trypsin, the experimental group showed a decrease in the percentage of G1-phase cells, but an increase in the percentage of S-phase cells, proliferation index, migratory and adhesive capacity compared with the control group （all P < 0.05）. Conclusion Trypsin can promote the proliferation, migration and adhesion of A431 cells.