siRNA干扰KLF8表达对鼻咽癌细胞上皮间质转化的作用

胡文兵; 王钢胜; 陈曦; 陈军; 於席芳

doi:10.3971/j.issn.1000-8578.2016.12.009

siRNA干扰KLF8表达对鼻咽癌细胞上皮间质转化的作用

Effect of Down-regulating KLF8 Expression by siRNA on EMT in Nasopharyngeal Carcinoma Cells

摘要

摘要:
目的探讨下调KLF8的表达对鼻咽癌CNE1-LMP1细胞侵袭能力的影响及其机制。
方法通过脂质体转染法将KLF8 siRNA真核表达质粒稳定转染至鼻咽癌细胞株CNE1-LMP1。Western blot和RT-PCR法检测CNE1-LMP1细胞中KLF8、E-cadherin、N-cadherin蛋白和mRNA表达水平的变化。Transwell侵袭实验观察siRNA后对CNE1-LMP1细胞侵袭能力的影响。
结果 KLF8 siRNA转染的CNE1-LMP1细胞株与其阴性对照NC-si组比较KLF8蛋白和mRNA表达水平均下调，证实稳转细胞株建立成功。KLF8 siRNA可上调CNE1-LMP1细胞中E-cadherin的表达，而下调N-cadherin的表达。Transwell侵袭实验显示siRNA干扰CNE1-LMP1细胞侵袭能力下降。
结论 KLF8在鼻咽癌CNE1-LMP1细胞中高表达；通过siRNA下调KLF8的表达能干扰上皮间质转化（EMT）相关蛋白E-cadherin、N-cadherin的表达，阻断EMT过程，抑制CNE1-LMP1细胞侵袭转移。

Abstract:
Objective To investigate the effect of down-regulating the expression of KLF8 by siRNA interference on the invasion of human nasopharyngeal carcinoma cells CNE1-LMP1 and related mechanism.
Methods The siRNA targeting KLF8 was designed and the eukaryotic expression plasmid was synthesized. The plasmid was transfected into CNE1-LMP1 cells. The expression of KLF8, E-cadherin and N-cadherin protein and mRNA were detected by Western blot and RT-PCR. Transwell invasion assay was employed to observe the invasion ability of CNE1-LMP1 cells through KLF8 siRNA.
Results The expression levels of KLF8 protein and mRNA were down-regulated in the KLF8-siRNA-transfected CNE1-LMP1 cell lines compared with negative control (NC-si) group, which verified that the stable transfected cell lines were successfully established. KLF8 siRNA could up-regulate the expression of E-cadherin and down-regulate the expression of N-cadherin in CNE1-LMP1 cells. Transwell invasion assay showed that siRNA decreased the invasion ability of CNE1-LMP1 cells.
Conclusion KLF8 is highly expressed in nasopharyngeal carcinoma cells CNE1-LMP1. KLF8 siRNA could affect the expression of E-cadherin and N-cadherin, and inhibit the invasion and metastasis of CNE1-LMP1 cells.

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