氯胺酮通过p38 MAPK信号通路对乳腺癌细胞生物学行为的影响

Ketamine inhibits proliferation,invasion and migration of breast cancer cells via p38 MAPK signaling pathway

目的:探讨氯胺酮通过p38 MAPK信号通路对乳腺癌细胞增殖、侵袭与迁移的影响。方法:以乳腺癌细胞MDA-MB-231为研究对象,根据细胞培养基添加氯胺酮和p38 MAPK抑制剂(SB203580)分为4组:control组(无氯胺酮和SB203580)、氯胺酮组(10μg/mL)、SB203580组(20μmol/L)、氯胺酮+SB203580组(氯胺酮10μg/mL+SB203580 20μmol/L);实时荧光定量PCR(RT-qPCR)法检测各组细胞中p38 MAPK磷酸化水平;克隆形成实验检测各组细胞的增殖能力;Transwell侵袭实验检测各组细胞的趋化运动能力和侵袭能力;细胞划痕实验检测各组细胞的迁移能力;RT-qPCR和Western blotting法检测各组细胞中MMP-2、MMP-9 mRNA和蛋白的表达。结果:与control组相比,氯胺酮组和SB203580组均能降低p38 MAPK磷酸化水平(P<0.05)以及抑制MDA-MB-231细胞增殖和侵袭能力(P<0.001或P<0.05),而与氯胺酮组或SB203580组相比,氯胺酮+SB203580组p38 MAPK磷酸化水平均降低(P<0.001),同时MDA-MB-231细胞增殖和侵袭能力降低(P<0.001);与control组相比,氯胺酮组和SB203580组中MDA-MB-231细胞迁移超出划痕部分减少(P<0.05),而与氯胺酮组或SB203580组相比,氯胺酮+SB203580组划痕距离均变宽(P<0.001);与control组相比,氯胺酮组和SB203580组MDA-MB-231细胞中MMP-2和MMP-9 mRNA和蛋白的表达降低(P<0.05),与氯胺酮组或SB203580组相比,氯胺酮+SB203580组MMP-2和MMP-9 mRNA和蛋白的表达均降低(均P<0.001)。结论:氯胺酮通过抑制p38 MAPK信号通路抑制乳腺癌细胞的增殖、侵袭与迁移。

Objective:To investigate the effect of ketamine on proliferation,invasion and migration of breast cancer cells through p38 MAPK signaling pathway.Methods:Taking breast cancer cells MDA-MB-231 as the research objects,they were divided into 4 groups:control group(without ketamine and SB203580),ketamine group(10 μg/mL),SB203580 group(20 μmol/L),and ketamine+SB203580 group(ketamine 10 μg/mL+SB203580 20μmol/L) according to the difference of adding ketamine(ketamine) and SB203580(p38 MAPK inhibitor) to the cell culture medium.RT-qPCR was used to detect the phosphorylation level of p38 MAPK.The colony formation assay was used to detect the proliferative capacity of cells.Transwell assay was used to detect the chemotactic activity and invasive ability of cells.The cell scratch test was used to detect the migration ability of cells.The mRNA and protein expressions of MMP-2 and MMP-9 were detected by RT-qPCR and western blotting,respectively.Results:Compared with the control group,the phosphorylation level of p38 MAPK,the proliferation and invasion of MDA-MB-231 cells were reduced in ketamine group and SB203580 group(P<0.05).Compared with ketamine group or SB203580 group,the phosphorylation level of p38 MAPK and the proliferation and invasion ability of cells in ketamine + SB203580 group were significantly reduced(P<0.001).Compared with the control group,the migration of cells was significantly reduced in ketamine group and SB203580 group(P<0.05).Compared with ketamine group or SB203580 group,the cells migration in ketamine+SB203580 group was significantly elevated(P<0.001).Compared with the control group,the expressions of MMP-2 and MMP-9 mRNA and protein in ketamine group and SB203580 group were decreased(P<0.05).Compared with ketamine group or SB203580 group,the expressions of MMP-2 and MMP-9 mRNA and protein were significantly reduced in ketamine+SB203580 group(P<0.001).Conclusion:Ketamine inhibits proliferation,invasion and migration of breast cancer cells by inhibiting p38 MAPK signaling pathway.