多功能蛋白聚糖对食管鳞癌细胞体外迁移和侵袭的影响

Effects of Versican on Migration and Invasion of Esophageal Squamous Cell Carcinoma Cell in Vitro

目的探讨多功能蛋白聚糖(versican,VCAN)在食管鳞癌(esophageal squamous cell carcinoma,ESCC)KYSE150和EC9706细胞中的表达及其对细胞体外迁移和侵袭的影响。方法从GEO数据库中下载GSE77861数据集,采用生物信息学方法进行数据处理并筛选差异表达基因,筛选出与ESCC预后相关的关键基因VCAN;Western blot检测人正常食管上皮细胞HEEC及4种ESCC细胞系KYSE70、KYSE140、KYSE150、EC9706中VCAN的蛋白表达水平。将KYSE150和EC9706细胞分3组(si-VCAN沉默表达组、si-NC阴性对照组和空白对照组)处理,然后Western blot检测各组VCAN的蛋白表达水平;划痕实验、Transwell实验分别检测沉默表达VCAN对细胞迁移和侵袭能力的影响。结果ESCC细胞系的VCAN相对表达水平高于人正常食管上皮细胞HEEC,且si-VCAN沉默表达组的VCAN表达水平低于si-NC阴性对照组和空白对照组,差异有统计学意义(P<0.05);划痕实验和Transwell实验结果表明,si-VCAN沉默表达组的迁移和侵袭细胞数目较对照组减少,差异有统计学意义(P<0.05)。结论VCAN在ESCC细胞系中高表达,沉默表达VCAN后可抑制ESCC细胞的体外迁移和侵袭。

Objective To explore the expression of Versican(VCAN)in esophageal squamous cell carcinoma(ESCC)KYSE150 and EC9706 cells and its impact on cell migration and invasion in vitro.Methods The GSE77861 data set was downloaded from the GEO database,bioinformatics methods were used to process the data and screen differentially expressed genes,and the key gene VCAN related to the prognosis of ESCC was screened out.Western blot was used to detect the protein expression levels of VCAN in human normal esophageal epithelial cells HEEC and four ESCC cell lines KYSE70,KYSE140,KYSE150,and EC9706.The treated KYSE150 and EC9706 cells were divided into 3 groups:si-VCAN silent expression group,si-NC negative control group and blank control group.Western blot was used to detect the protein expression level after silent expression of VCAN,wound healing assay was used to detect the effect of silent expression of VCAN on cell migration ability,and Transwell assay was applied to detect the effect of silent expression of VCAN on cell invasion capacity.Results Western blot results showed that the relative expression level of VCAN in ESCC cell lines was higher than that in human normal esophageal epithelial cells HEEC.The si-VCAN silent expression group’s VCAN expression was less than that of the si-NC negative control and the blank control group(P<0.05).The wound healing and transwell assay results showed that the number of migrating and invading cells in the si-VCAN silenced expression group was reduced compared to the control group(P<0.05).Conclusion VCAN is highly expressed in ESCC cell lines,and silencing the expression of VCAN can inhibit the migration and invasion of ESCC cells in vitro.