大黄素对口腔鳞状细胞癌增殖、侵袭和迁移的影响

Effect of Emodin on Proliferation,Invasion and Migration of Oral Squamous Carcinoma Cells

目的:探讨大黄素对人口腔鳞状细胞癌CAL-27及SCC-15的增殖、侵袭和迁移能力的影响及作用机制。方法:采用不同浓度的大黄素处理CAL-27及SCC-15细胞,使用CCK-8法检测其增殖活性,根据结果计算不同作用时间细胞抑制率为50%时对应的药物浓度(IC50);划痕实验、Transwell侵袭实验分别检测大黄素对两种细胞迁移、侵袭能力的影响;蛋白免疫印迹法(Western blot)检测大黄素对细胞中金属基质蛋白酶-2(MMP-2)和金属基质蛋白酶-9(MMP-9)蛋白表达水平的影响;实时定量PCR(qRT-PCR)检测大黄素对细胞中MMP-2和MMP-9 mRNA表达水平的影响。结果:与对照组相比,不同浓度的大黄素均可抑制CAL-27和SCC-15细胞的增殖能力(P<0.05),且呈时间-浓度依耐性;与对照组相比,大黄素可明显降低两种细胞划痕的愈合率和侵袭细胞数(P<0.05),具有浓度依耐性;与对照组相比,经大黄素处理的两种细胞中MMP-2和MMP-9 mRNA及蛋白的表达水平均明显下调(P<0.05)。结论:大黄素在体外可以显著抑制人OSCC细胞的增殖、侵袭和迁移能力,其作用可能与下调MMP-2、MMP-9的表达有关。

Objective:To investigate the effect of emodin on proliferation,invasion,and migration of human oral squamous cell carcinoma cells CAL-27 and SCC-15 and its molecular mechanism.Methods:CAL-27 and SCC-15 cells were treated with different concentrations of emodin.CCK-8 was used to detect their proliferation activity.According to the results,the corresponding drug concentration(IC50)was calculated when the cell inhibition rate was 50%at different time.Scratch test and Transwell invasion test were used to detect the effects of emodin on the migration and invasion of two kinds of cells.Western blot was used to detect the effect of emodin on the expression of MMP-2 and MMP-9.Real time quantitative PCR(qRT-PCR)was used to detect the effect of emodin on the expression of MMP-2 and MMP-9 mRNA.Results:Compared with the control group,emodin at different concentrations could inhibit the proliferation of CAL-27 and SCC-15 cells(P<0.05)in a time concentration dependent manner.Compared with the control group,Rhein could significantly reduce the wound healing rate and the number of invasive cells(P<0.05),which was concentration dependent.Compared with the control group,the mRNA and protein expression levels of MMP-2 and MMP-9 in emodin treated cells were significantly decreased(P<0.05).Conclusion:Emodin can significantly inhibit the proliferation,invasion,and migration of human oral squamous cell carcinoma cells in vitro,which may relate to the down regulation of the expression of MMP-2 and MMP-9.