半乳凝素3对结直肠癌细胞生物学行为的影响

Effect of Galectin-3 on biological behavior of colorectal cancer cells

目的观察半乳凝素3(Galectin-3)对结直肠癌细胞的增殖、迁移及成瘤的影响。方法在肠癌细胞株SW480细胞中采用CRISPR-Cas9技术建立Galectin-3敲除(KO)的稳定细胞株,并以未敲除Galectin-3的细胞作为对照组。采用蛋白质印迹法(Western blot)鉴定对照组和Galectin-3 KO组细胞Galectin-3表达水平;采用Edu法分析对照组和Galectin-3 KO组细胞的增殖能力;采用划痕实验和Transwell实验分析对照组和Galectin-3 KO组细胞迁移能力;采用体内成瘤实验分析对照组和观察组细胞肿瘤生长。结果选取两条单导RNA(sgRNA)靶向敲除Galectin-3蛋白,分别获得了Galectin-3 KO1和Galectin-3 KO2细胞株。与对照组[(52.19±12.09)%]比较,Galectin-3 KO1组和Galectin-3 KO2组细胞的EDU阳性率[(11.35±9.27)%和(14.39±7.88)%]显著下降,差异有统计学意义(t=3.912,P<0.05;t=3.209,P<0.05)。与对照组[(81.32±9.81)%]比较,Galectin-3 KO1组和Galectin-3 KO2组细胞划痕愈合距离百分比[(33.28±8.77)%和(30.19±9.10)%]显著下降,差异有统计学意义(t=5.197,P<0.05;t=5.273,P<0.05)。与对照组[(201.33±19.81)个]比较,Galectin-3 KO1组和Galectin-3 KO2组平均每个视野细胞迁移数量[(109.41±12.87)个和(125.10±15.91)个]显著下降,差异有统计学意义(t=9.255,P<0.05;t=10.364,P<0.05)。Galectin-3 KO1组和Galectin-3 KO2组细胞在裸鼠体内生长速度明显低于对照组,差异有统计学意义(P<0.05)。结论检测Galectin-3蛋白水平有助于判断结直肠细胞增殖和迁移。

Objective To investigate the effects of Galectin-3 on the proliferation,migration and tumorigenesis of colorectal cancer cells.Methods The stable cell line of Galectin-3 knockout(KO)was established by CRISPR-Cas9 technique in SW480 cells and the cells without knockout of Galectin-3 were used as control group.The expression level of Galectin-3 in control group and Galectin-3 KO group were analyzed by Western blotting.The proliferation ability in control group and Galectin-3 KO group were analyzed by Edu method.The migration ability in control group and Galectin-3 KO group were analyzed by scratch test and Transwell experiment.and was used to analyze The growth of tumors cell in control group and Galectin-3 KO group were analyzed by in vivo tumorigenesis experiment.Results Two single guide RNA(sgRNA)targeting knockout Galectin-3 proteins were selected and Galectin-3 KO1 and Galectin-3 KO2 were obtained respectively.Compared with the control group[(52.19±12.09)%],the positive rate of EDU in the Galectin-3 KO1 group and Galectin-3 KO2 groups[(11.35±9.27)%and(14.39±7.88)%]decreased significantly(t=3.912,P<0.05;t=3.209,P<0.05).Compared with the control group[(81.32±9.81)%],the percentage of healing distance of scratches in Galectin-3 KO1 group and Galectin-3 KO2 group[(33.28±8.77)%and(30.19±9.10)%]decreased significantly(t=5.197,P<0.05;t=5.273,P<0.05).Compared with the control group[(201.33±19.81)cells],the average number of cell migration per visual field in Galectin-3 KO1 group and Galectin-3 KO2 group[(109.4±12.87)cells and(125.10±15.91)cells]decreased significantly(t=9.255,P<0.05;t=10.364,P<0.05).The growth rate of cells in Galectin-3 KO1 group and Galectin-3 KO2 group in nude mice was significantly lower than that in control group(P<0.05).Conclusion Detection of Galectin-3 protein level is helpful to judge the proliferation and migration of colorectal cells.