南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (11): 1643-.

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Kiss-1基因通过NF-κB信号传导通路抑制人结直肠癌HCT116细胞迁移

陈绍勤,苏小宝,高骥,韩宏景,陈志华,林素勇   

  • 出版日期:2015-11-20 发布日期:2015-11-20

Suppression of Kiss-1 gene inhibits HCT116 human colorectal carcinoma cell migration
in vitro via nuclear factor-κB signaling pathway

  • Online:2015-11-20 Published:2015-11-20

摘要: 目的探讨Kiss-1 基因抑制人结直肠癌细胞转移能力与NF-κB信号传导通路的相关性。方法构建重组pGC-LV-Kiss-
1-EGFP慢病毒载体并转染人结直肠癌HCT116细胞,实验分为空白对照组(CON组)、慢病毒空载体阴性对照组(NC组)、Kiss-1
基因过表达组(OE组)。CCK-8(Cell Counting Kit-8)法检测转染前后细胞增殖能力的变化,Transwell法分别检测转染前后细
胞的侵袭和迁移能力的变化。Western-blot法检测转染前后NF-κB信号传导通路中抑制性蛋白I-κB以及下游效应蛋白MMP-9
表达量的变化。结果OE组HCT116细胞中I-κB的表达含量较CON组、NC组明显升高(P<0.05),下游效应蛋白MMP-9的表
达量明显下降(P<0.05),差异具有统计学意义。OE组较CON组、NC组细胞增殖受到明显抑制(P<0.05),细胞侵袭、迁移能力
亦出现明显抑制(P<0.05)。结论重组pGC-LV-Kiss-1-EGFP慢病毒转染人结直肠癌HCT116细胞后,可能通过NF-κB信号传
导通路途径抑制其增殖、侵袭和迁移能力。

Abstract: Objective To investigate the effect of Kiss-1 gene suppression on the metastatic capacity of HCT116 human
colorectal carcinoma cells in vitro and the involvement of nuclear factor-κB (NF-κB) signaling pathway. Methods A
recombinant lentiviral vector of Kiss-1 gene pGC-LV-Kiss-1-EGFP or the empty vector was transfected in HCT116 cells. Cell
Counting Kit-8 (CCK8) and Transwell chamber assay were used to detect the changes in cell proliferation, invasion and
migration ability after the transfection. Western blotting was used to detect the expression of I-κB, the inhibitive protein of
NF-κB signal pathway, and the expression of the downstream effector MMP-9 before and after transfection. Results In cells
over-expressing Kiss-1, I-κB expression increased and MMP-9 expression decreased significantly compared to those in the
blank control and vector-transfected cells (P<0.05). Kiss-1 gene over-expression resulted in significant inhibition of HCT116 cell
proliferation, invasion, and migration as compared to the control cells (P<0.05). Conclusion Lentivirus-mediated Kiss-1 gene
over-expression can inhibit the proliferation, invasion, and migration of HCT116 cells via the NF-B signaling pathway.