Effect of Twist on angiogenesis of nasopharyngeal cancer cells by regulating ERK signaling pathway / 中国病理生理杂志

ABSTRACT

AIM:To investigate the effect of Twist transcription factor on angiogenesis of nasopharyngeal cancer cells by regulating ERK signaling pathway in vitro.METHODS:Twist siRNA was transfected into the CNE2 cells with Lipofectamine 2000, and the CNE2 cell-conditioned medium (CM) was prepared, which was then used to treat human umbilical vein endothelial cells (HUVECs).The CCK-8 assay, Boyden chamber and capillary tube formation assay were used to evaluate the angiogenic activity of HUVECs.The mRNA expression of Twist in nasopharyngeal cancer cells was detected by real-time PCR.The protein levels of Twist, ERK, p-ERK, p38, p-p38, JNK, p-JNK and VEGF were detected by Western blot.The ERK, p38 and JNK signaling pathway inhibitors were employed to treat HUVECs, and their effect on the capillary tube formation of HUVECs was analyzed.RESULTS:The expression of Twist at mRNA and protein levels in the CNE2 cells was significantly inhibited after transfection with Twist siRNA (P<0.01).The abilities of proliferation, invasion and capillary tube formation of HUVECs were markedly suppressed after knockdown of Twist expression (P<0.01).The protein level of p-ERK was significantly elevated in the CNE2-siTwist cells, while VEGF expression was decreased.Knockdown of Twist did no change the levels of p-p38 and p-JNK.The ERK signaling pathway inhibitor partly abrogated the anti-angiogenic effect of Twist knockdown.CONCLUSION:Knockdown of Twist inhibits the angiogenesis of nasopharyngeal cancer cells in vitro partly through activation of ERK signaling pathway.