下调miR-34a表达逆转七氟醚对食管癌KYSE-150细胞增殖及侵袭的抑制作用

Down-regulation of microRNA-34a reverses inhibitory effect of sevoflurane on proliferation and invasion of esophageal cancer KYSE-150 cells

目的:探讨下调微RNA(microRNA,miR)-34a对七氟醚抑制食管癌KYSE-150细胞增殖、侵袭及迁移能力的影响。方法:用不同体积百分浓度(1.7%、3.4%和5.1%)的七氟醚处理KYSE-150细胞2、4和6 h后,采用MTT法检测各组细胞的增殖活性,实时荧光定量PCR法检测KYSE-150细胞中miR-34a的表达量。分别将携带有miR-34a-抑制子(miR-34a-inhibitor)及阴性对照(negative control,NC)-抑制子(NC-inhibitor)的重组质粒转入KYSE-150细胞,实时荧光定量PCR法检测miR-34a表达量的改变情况。用5.1%七氟醚处理miR-34a沉默表达后的KYSE-150细胞,分别采用MTT法及Transwell小室法检测各组KYSE-150细胞的增殖、迁移及侵袭情况。结果:与未用七氟醚处理的对照组相比,不同体积百分浓度(1.7%、3.4%和5.1%)的七氟醚处理KYSE-150细胞2、4和6 h后,KYSE-150细胞的增殖被明显抑制(P值均<0.05),且随着七氟醚浓度的提高miR-34a的表达水平明显上调(P值均<0.05)。与转染NC-inhibitor的对照组相比,转染miR-34a-inhibitor后明显降低了KYSE-150细胞中miR-34a的表达水平(P <0.05);沉默miR-34表达逆转了七氟醚对KYSE-150细胞增殖、迁移和侵袭的抑制作用(P值均<0.05)。结论:下调miR-34a可逆转七氟醚对KYSE-150细胞生物学特性的抑制作用。

Objective: To investigate the effects of down-regulating microRNA(microRNA, miR)-34 a expression on the proliferation, invasion and migration of esophageal cancer KYSE-150 cells regulated by sevoflurane.Methods: KYSE-150 cells were treated with different concentrations of sevoflurane(1.7%, 3.4% and 5.1%) for 2, 4 and 6 h, then the cell proliferation was detected by MTT assay, while the expression level of miR-34 a was detected by real-time fluorescent quantitative PCR. The recombinant plasmids carrying miR-34 ainhibitor or negative control(NC)-inhibitor were transfected into KYSE-150 cells, then the expression level of miR-34 a was detected by real-time fluorescent quantitative PCR. The miR-34 a-inhibitor transfected KYSE-150 cells were treated with 5.1% sevoflurane, then the cell proliferation, migration and invasive abilities were detected by MTT and Transwell chamber method, respectively.Results: Compared with the control group, the different concentrations of sevoflurane(1.7%,3.4% and 5.1%) treatment for 2, 4 and 6 h inhibited the proliferation of KYSE-150 cells(all P < 0.05), and the expression level of miR-34 a was significantly increased by a dosedependent manner(all P < 0.05). Compared with the NC-inhibitor group, the expression level of miR-34 a in KYSE-150 cells transfected with miR-34 a-inhibitor was significantly decreased(P < 0.05). The knock-down of miR-34 a expression improved the proliferation,invasion and migration of KYSE-150 cells(all P < 0.05), reversing the inhibitory effects of sevoflurane.Conclusion: Down-regulation of miR-34 a reverses the inhibitory effects of sevoflurane in regulating the biological characteristics of KYSE-150 cells.