摘要
目的:探讨汉黄芩素对人口腔鳞状细胞癌SCC-4细胞生长和侵袭的影响及其作用机制。方法:使用不同浓度(0、20、40、60、80和100 mg/L)的汉黄芩素处理SCC-4细胞不同时间,分别用MTT法检测细胞活力,流式细胞术及Annexin V/PI双染法检测细胞凋亡,Transwell小室检测细胞侵袭能力,Western blot法检测Wnt/β-catenin信号通路的活化。结果:汉黄芩素呈剂量及时间依赖性地抑制细胞生长,同时诱导细胞大量凋亡,抑制细胞的侵袭。汉黄芩素明显抑制了细胞中β-catenin的活化,同时其下游靶分子细胞周期蛋白D1(cyclin D1)、基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)和MMP-9的表达水平降低,而抗凋亡蛋白Bcl-2的表达增加。用Wnt/β-catenin通路激动剂Li Cl处理后,汉黄芩素抑制的Wnt/β-catenin通路分子活化明显增强,同时细胞生长能力明显增强,而凋亡能力降低,还明显减弱了汉黄芩素对细胞侵袭能力的抑制作用。结论:汉黄芩素主要通过抑制Wnt/β-catenin信号通路来调控口腔鳞状细胞癌细胞的生长及侵袭进程,具有一定的抗口腔鳞状细胞癌发展的作用,可成为临床上口腔鳞状细胞癌防治的潜在药物。
AIM:To investigate the molecular mechanism of wogonin on the growth and invasion of oral squa -mous-cell carcinoma (OSCC) cell line SCC-4.METHODS:After treatment with various doses (0, 20, 40, 60, 80 and 100 mg/L) of wogonin for the indicated time , MTT assay was used to evaluate cell viability .The cell apoptosis was detec-ted by flow cytometry with Annexin V and propidium iodide (Annexin V/PI) double staining.The cell invasion ability was analyzed by Transwell assay .The activation of Wnt/β-catenin signaling molecules was assessed by Western blot .RE-SULTS:Wogonin inhibited the viability and invasion of SCC-4 cells but promoted cell apoptosis in a dose-and time-de-pendent manner .Wogonin treatment obviously decreased the activation of β-catenin.Moreover, the expression of down-stream targets cyclin D1, matrix metalloproteinase-2 (MMP-2) and MMP-9 were obviously down-regulated, accompanied by the up-regulation of anti-apoptotic protein Bcl-2.Wnt/β-catenin activator LiCl remarkably attenuated the inhibitory effect of wogonin on the activation of Wnt/β-catenin signaling molecules .Importantly, the inhibition of cell growth and in-vasion ability by wogonin treatment was dramatically attenuated after LiCl exposure .CONCLUSION: Wogonin blocks SCC-4 cell growth and invasion mainly by regulating Wnt/β-catenin signaling , indicating that it is a potential suppressor for OSCC and may be a potential target for the development of anti-OSCC therapy .
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2017年第10期1869-1873,共5页
Chinese Journal of Pathophysiology
基金
甘肃省城市发展研究院科研项目(No.2014-GSCFY-KJ07)