microRNA-21通过上皮间质转化促进胆管癌细胞侵袭转移的研究

MicroRNA-21 promoting the invasion and metastasis of cholangiocarcinoma through epithelial-mesenchymal transition

目的探讨micro RNA-21(miR-21)对胆管癌RBE细胞侵袭转移能力的影响及其内在机制。方法 miR-21及其抑制基因片段(miR-21i)通过慢病毒载体感染胆管癌RBE细胞,RT-PCR用于检测RBE细胞中miR-21表达;Transwell体外侵袭实验和细胞划痕试验用于检测RBE细胞侵袭及转移能力;Western blotting用于检测PTEN蛋白和上皮间质转化(EMT)特征性蛋白E-cadherin、N-cadherin、Vimentin表达情况。结果 miR-21过表达促进RBE细胞侵袭和转移,miR-21低表达抑制RBE细胞的侵袭和转移。miR-21过表达能够降低PTEN蛋白和E-cadherin蛋白,增强N-cadherin和Vimentin蛋白的表达;转染miR-21i抑制基因组(miR-21i)组则有相反的效果,而转染空白基因组(对照组)相应蛋白表达无明显变化。结论 miR-21起到癌基因的作用,可增强胆管癌细胞侵袭和转移能力,其机制是通过调节抑癌基因PTEN及改变EMT相关蛋白的变化。这些发现为胆管癌侵袭转移的分子机制提供新的科学依据。

Objective To investigate the effect of micro RNA-21（miR-21） on the invasion and metastasis of cholangiocarcinoma RBE cells, as well as its underlying mechanisms. Methods Cholangiocarcinoma RBE cells were transfected miR-21 or miR-21 inhibitor（miR-21 i） mediated in lentivirus. Reverse transcription polymerase chain reaction（RT-PCR） was used to examine the expression of miR-21. Transwell chambers and wound-healing assay were applied to measure the invasive and metastatic effects of RBE cells. Western blotting analysis was used to assess the expression of PTEN protein and epithelial-mesenchymal transition（EMT） related proteins, including E-cadherin, N-cadherin, and Vimentin. Results The over-expression of miR-21 promoted invasion and metastasis of cholangiocarcinoma RBE cells, miR-21 inhibitor suppressed these effects. Furthermore, it has been found that the over-expression of miR-21 up-regulated PTEN and E-cadherin proteins, and down-regulated N-cadherin and Vimentin proteins. However, these data were not observed in the control group. Conclusions miR-21 acts as an oncogene, promotes invasion and metastasis of cholangiocarcinoma cells through PTEN and EMT. These findings provide new scientific basis.