盘状结构域受体1通过诱导上皮间质转化促进胰腺癌细胞迁移和侵袭

Discoidin domain receptor 1 promoting migration and invasion of pancreatic cancer cells by inducing epithelial mesenchymal transformation

目的探讨盘状结构域受体1(DDR1)调控上皮间质转化(EMT)对胰腺癌细胞PANC-1迁移侵袭及细胞外基质降解能力的影响。方法检测人正常胰腺导管上皮细胞H6C7及胰腺癌细胞株(PANC-1、AsPC-1、BxPC-3、SW1990)中的DDR1表达水平。在PANC-1细胞中转染DDR1表达质粒,通过Western blot检测DDR1的蛋白表达,明确其转染效果。细胞划痕实验和Transwell检测DDR1表达水平升高对细胞迁移侵袭的影响。原位明胶酶谱法检测DDR1过表达对细胞外基质降解能力的影响。Western blot检测EMT标记蛋白E-cadherin、Vimentin和α-SMA的表达;qRT-PCR检测转录因子Snail1、Snail2、Twist和Zeb1的表达。结果与对照组比较,胰腺癌细胞株中DDR1基因表达增加(P<0.05)。在PANC-1细胞中过表达DDR1后,其迁移侵袭能力及明胶基质降解能力均增强(P<0.05)。与对照组比较,实验组E-cadherin蛋白表达降低,Vimentin和α-SMA蛋白表达升高,转录因子Snail2和Twist基因表达升高(均P<0.05)。但2组的Snail1和Zeb1基因表达差异无统计学意义(P>0.05)。结论DDR1在胰腺癌细胞中高表达,可能通过上调Snail2和Twist的基因表达诱导EMT,促进PANC-1细胞迁移侵袭。

Objective To investigate the effects of epithelial mesenchymal transformation(EMT)regulated by discoidin domain receptor 1(DDR1)on migration,invasion and extracellular matrix degradation of pancreatic cancer cell PANC-1.Methods The expression level of DDR1 in human normal pancreatic duct epithelial cells H6 C7 and pancreatic cancer cell lines(PANC-1,AsPC-1,BxPC-3,SW1990)was detected.The expression plasmid of DDR1 was transfected into PANC-1 cells,and the protein expression of DDR1 was detected by Western blot to determine its transfection effect.The effect of increased DDR1 expression level on cell migration and invasion were detected by cell scratch assay and Transwell.The effect of DDR1 overexpression on extracellular matrix degradation was detected by in situ gelatin zymography.The expressions of EMT labelled proteins E-cadherin,Vimentin andα-SMA were detected by Western blot.qRT-PCR was used to detect the expression of transcription factors Snail1,Snail2,Twist and Zeb1.Results Compared with the control group,the expression of DDR1 gene in pancreatic cancer cell lines increased(P<0.05).After the overexpression of DDR1 in PANC-1 cells,the migration and invasion ability and degradation ability of gelatin matrix were enhanced(P<0.05).Compared with the control group,the expression of E-cadherin protein in the experimental group decreased,the expression of Vimentin andα-SMA protein increased,and the expression of transcription factors Snail2 and Twist genes increased(all P<0.05).However,there was no significant difference in the expression of Snail1 and Zeb1 gene between the two groups(P>0.05).Conclusion DDR1 is highly expressed in pancreatic cancer cells,which may induce EMT by up-regulating the expression of Snail2 and Twist genes and promote the migration and invasion of PANC-1 cells.