HIF-1α通过调控上皮-间充质转化对胆囊癌侵袭转移的影响

The Effect of HIF-1αon Invasion and Metastasis of Gallbladder Cancer by Regulating the Epithelial-Mesenchymal Transformation

目的探讨HIF-1α对胆囊癌细胞迁移侵袭的影响及其相关调控机制。方法构建GBC-SD细胞,使用慢病毒转染,分别得到实验组(HIF-1αshRNA)、阴性对照组(空载慢病毒转染)和空白对照组(未处理)。蛋白质免疫印迹法检测E-Cadherin、N-Cadherin蛋白的表达水平,GBC-SD细胞的迁移能力用细胞划痕实验检测,GBC-SD细胞的侵袭能力用Transwell侵袭实验检测。结果与对照组相比,实验组E-Cadherin蛋白的表达水平明显升高(P<0.001),N-Cadherin蛋白的表达水平明显降低(P<0.001),对照组之间E-Cadherin、NCadherin蛋白的表达水平无明显差异(P>0.05)。实验组细胞划痕愈合度较对照组低(P<0.001)、穿模个数较对照组少(P<0.001),而对照组之间细胞划痕愈合度、穿模个数无明显差异(P>0.05)。结论靶向敲低HIF-1α可以降低GBC-SD细胞的迁移侵袭能力,其作用机制可能与调控上皮-间充质转化有关。

Objective To explore the effect of HIF-1αon the migration and invasion of gallbladder cancer cells and its related regulatory mechanism.Methods GBC-SD cells were constructed and transfected with lentivirus,and the experimental group(HIF-1αshRNA),negative control group(empty lentivirus transfection)and blank control group(untreated)were obtained respectively.The expression levels of E-Cadherin,and NCadherin proteins were detected by Western blotting assay,Cell scratch assay was ued to detect the migratory ability of GBC-SD cells,and Transwell invasion assay was uesd to detect the invasive ability of GBC-SD cells.Results Compared with the control group,the expression level of E-Cadherin protein in the experimental group was significantly increased(P<0.001),while the expression level of N-Cadherin protein was significantly decreased(P<0.001),and there was no remarkable difference in the expression level of E-Cadherin and N-Cadherin proteins between the control groups(P both>0.05).In the experimental group,the cell scratch healing degree was inferior to the control group(P<0.001),and the number of penetrating molds was also inferior to the control group(P<0.001),while there was no obvious otherness in the cell scratch healing degree and the number of penetrating molds between the control groups(P>0.05).Conclusion Targeted knockdown of HIF-1αcan reduce the migration and invasion ability of gallbladder cancer cells,and its mechanism of action may be related to the regulation of epithelial mesenchymal transition.