七氟醚调控miR-34a/HMGB1表达抑制乳腺癌MDA-MB-231细胞增殖和侵袭

Effects of sevoflurane on proliferation and invasion of breast cancer MDA-MB-231 cells and its action mechanism

目的探讨七氟醚对乳腺癌MDA-MB-231细胞增殖、侵袭的影响及其机制。方法以1.7%、3.4%和5.1%七氟醚刺激MDA-MB-231细胞4 h后,采用MTT法、克隆形成实验和Transwell小室实验分别检测细胞的增殖、克隆形成和侵袭能力,RT-PCR检测细胞中miR-34a和HMGB1 mRNA的表达,Western blot检测HMGB1蛋白的表达。采用荧光素酶实验检测miR-34a和HMGB1的靶向关系,脂质体法转染miR-34a抑制剂后,观察下调miR-34a表达对5.1%七氟醚刺激下的MDA-MB-231细胞增殖、侵袭和HMGB1表达的影响。结果七氟醚能够呈浓度依赖性抑制MDA-MB-231细胞的增殖、克隆形成和侵袭,并上调miR-34a、下调HMGB1mRNA和蛋白的表达。HMGB1是miR-34a的靶基因。转染miR-34a抑制剂成功下调miR-34a表达后,七氟醚对MDA-MB-231细胞增殖、克隆形成和侵袭能力以及对HMGB1表达的抑制作用均明显得以逆转。结论七氟醚可通过调控miR-34a/HMGB1表达抑制MDA-MB-231细胞增殖和侵袭。

Objective To investigate the effects of sevoflurane on proliferation and invasion of breast cancer MDA-MB-231 cells and its action mechanism.Methods The MDA-MB-231 cells were stimulated with 1.7%,3.4%and 5.1%sevoflurane for 4 hours.The proliferation,clone formation and invasion ability of MDA-MB-231 cells were detected by MTT,clone formation test and Transwell chamber assay,respectively.The expression levels of miR-34a and HMGB1 were detected by RT-PCR,and the expression levels of HMGB1 protein were detected by Western Blot.Luciferase assay was used to detect the targeting correlation between miR-34a and HMGB1.Moreover the effects of down-regulation of miR-34a expression on proliferation,invasion and HMGB1 expression of MDA-MB-231 cells stimulated by 5.1%sevoflurane were observed after transfection of miR-34a inhibitors by liposome.Results Sevoflurane could inhibit the proliferation,cloning and invasion of MDA-MB-231 cells in a concentration-dependent manner,and up-regulated the expression levels of miR-34a as well as HMGB1 gene and protein.And HMGB1 was the target gene of miR-34a.The inhibition of sevoflurane on the proliferation,cloning and invasion of MDA-MB-231 cells and the expression of HMGB1 were reversed after the successful down-regulation of the expression of miR-34a by the transfected inhibitor of miR-34a.Conclusion Sevoflurane can inhibit the proliferation and invasion of MDA-MB-231 cells by regulating the expression of miR-34a/HMGB1.