miR-204通过靶向调控SOX4抑制急性淋巴细胞白血病CEM/C1细胞增殖、侵袭的体外实验研究

Inhibition effect of miR-204 on proliferation,migration and invasion of acute lymphoblastic leukemia CEM/C1 cells by targeting SOX4

目的:探讨miR-204对急性淋巴细胞白血病(ALL)CEM/C1细胞的增殖、迁移和侵袭的影响及其与SOX4的靶向关系。方法:选取ALL细胞株CEM/C1,对CEM/C1细胞进行转染,分为NC组(无处理)、miR-NC组(转染miR-NC)、miR-204组(转染miR-204-mimic)、si-NC组(转染si-NC)、si-SOX4组(转染si-SOX4)、miR-204+pcDNA组(转染miR-204-mimic+pcDNA)及miR-204+pcDNA-SOX4组(转染miR-204-mimic+pcDNA-SOX4)。采用实时荧光定量PCR法检测细胞中miR-204表达水平,MTT检测细胞增殖能力,Transwell小室实验检测细胞迁移、侵袭能力,Western blot检测细胞SOX4表达水平,流式细胞仪检测细胞凋亡,荧光素酶实验验证miR-204与SOX4的靶向关系。结果:转染细胞48 h和72 h时,miR-204组细胞活力、迁移和侵袭数量均明显低于NC组、miR-NC组(P<0.05),凋亡率高于NC组、miR-NC组(P<0.05)。si-SOX4组细胞活力、迁移和侵袭数量均明显低于NC组、si-NC组(P<0.05),凋亡率高于NC组、si-NC组(P<0.05)。miR-204+pcDNA-SOX4组细胞48 h、72 h时细胞活力、细胞迁移和侵袭数量均明显高于miR-204+pcDNA组、miR-204组(P<0.05),凋亡率低于miR-204+pcDNA组、miR-204组(P<0.05)。与对照组相比,SOX4野生型质粒组荧光素酶活性明显降低(P<0.05),SOX4突变型质粒组荧光素酶活性无明显变化(P>0.05)。结论:miR-204可负性调控SOX4抑制急性淋巴细胞白血病CEM/C1细胞增殖、迁移和侵袭。

Objective:To investigate the effect of miR-204 on the proliferation,migration and invasion of acute lymphoblastic leukemia(ALL)CEM/C1 cells and its targeting relationship with SOX4.Methods:ALL cell lines CEM/C1 was selected.According to the different transfection substances,they were divided into NC group(CEM/C1 cells),miR-NC group(miR-NC),miR-204 group(miR-204-mimic),si-NC group(transfected with si-NC),si-SOX4 group(transfected with si-SOX4),miR-204+pcDNA group(transfected with miR-204-mimic+pcDNA)and miR-204+pcDNA-SOX4 group(transfected with miR-204-mimic+pcDNA-SOX4).Real time fluorescent quantitative PCR was used to detect the expression of miR-204.MTT was used to detect cell proliferation.Transwell chamber test was used to detect cell migration and invasion.Western blot was used to detect the expression level of SOX4.Luciferase assay was used to verify the relationship between miR-204 and SOX4.Results:At 48 h and 72 h,the cell viability,migration and invasion of miR-204 group were significantly lower than those of NC group and miR-NC group(P<0.05),apoptosis rate was higher than those of NC group and miR-NC group(P<0.05).At 48 h and 72 h,the cell viability,migration and invasion in si-SOX4 group were significantly lower than those in NC group and si-NC group(P<0.05),apoptosis rate was higher than those of NC group and si-NC group(P<0.05).The cell viability,migration and invasion of miR-204+pcDNA-SOX4 group were higher than those of miR-204+pcDNA group and miR-204 group at 48 h and 72 h(P<0.05),apoptosis rate was lower than those of miR-204+pcDNA group and miR-204 group(P<0.05).The luciferase reporter assay showed that miR-204 could reduce the activity of luciferase in SOX4 wild-type vectors(P<0.05).But there was no statistically significant difference in SOX4 mutant vectors(P>0.05).Conclusion:miR-204 can negatively regulate SOX4 and inhibit the proliferation,migration and invasion of ALL CEM/C1 cells.