SCO2介导乳腺癌MDA-MB-231细胞迁移浸润的机制研究

Mechanism of SCO2-mediated migration and infiltration of breast cancer MDA-MB-231 cells

目的探究SCO2介导乳腺癌MDA-MB-231细胞迁移浸润的机制。方法以乳腺癌细胞系MDA-MB-231作为研究对象,通过瞬时转染干扰其SCO2并分为对照组(si-NC组)和干扰组(si-SCO2组),采用实时荧光定量聚合酶链反应和Western blotting检测SCO2干扰效果,通过往siSCO2组细胞加入正常MDA-MB231细胞分泌的小细胞外囊泡的方式挽救SCO2,并分为对照组(siNC组)、干扰组(si-SCO2组)和挽救组(si-SCO2+NC-sEVs组);进一步构建SCO2过表达质粒并将质粒转染至细胞中并分为对照组(OE-NC组)和过表达组(OE-SCO2组),采用划痕试验和Transwell浸润试验分别检测各组迁移能力和侵袭能力。结果si-SCO2组SCO2 mRNA、蛋白相对表达量均较si-NC组细胞低(P<0.05)。si-SCO2组细胞迁移率较si-NC组低(P<0.05),细胞侵袭数较si-NC组少(P<0.05)。si-SCO2组细胞迁移率较si-NC组低,细胞侵袭数较si-NC组少(P<0.05),si-SCO2+NC-sEV组细胞侵袭数较si-SCO2组高(P<0.05)。OE-SCO2组SCO2 mRNA、蛋白相对表达量较OE-NC组高(P<0.05)。OE-SCO2组细胞迁移率较OE-SCO2组高(P<0.05),细胞侵袭数较OE-SCO2组多(P<0.05)。结论敲低SCO2能降低MDA-MB-231细胞的迁移能力和浸润能力,过表达SCO2可促进乳腺癌MDA-MB-231细胞的迁移能力和浸润能力,SCO2有望成为治疗乳腺癌转移的调控靶点。

Objective To explore the mechanism of SCO2 in mediating the migration and infiltration of breast cancer MDA-MB-231 cells.Methods The breast cancer cell line MDA-MB-231 was used as the model,and transient transfection to interfere with the expression of SCO2 was performed,where cells were divided into a control group(si-NC group)and an interference group(si-SCO2 group).The effect of SCO2 interference was detected by quantitative real-time polymerase chain reaction and Western blotting.Expression of SCO2 was rescued by adding small extracellular vesicles secreted by untreated MDA-MB231 cells to the cells in the si-SCO2 group,and the cells were divided into a control group(si-NC group),an interference group(si-SCO2 group)and a rescue group(si-SCO2+NC-sEVs group)based on the treatment.The SCO2 overexpression plasmid was further constructed and transfected into cells,and the control group(OE-NC group)and the overexpression group(OESCO2 group)were established.All groups of cells were tested for the migration and invasion ability by the scratc assay and the Transwell invasion assay,respectively.Results The relative mRNA and protein expressions of SCO2 in the si-SCO2 group were lower than those in the si-NC group(P<0.05).The cell migration rate in the si-SCO2 group was lower than that in the si-NC group(P<0.05),and the number of invasive cells in the si-SCO2 was lower than that in the si-NC group(P<0.05).The number of invasive cells in the si-SCO2+NC-sEV group was higher than that in the si-SCO2 group(P<0.05).The relative mRNA and protein expressions of SCO2 in the OE-SCO2 group were higher than those in the OE-NC group(P<0.05).The cell migration rate in the OE-SCO2 group was higher than that in the OE-NC group(P<0.05),and that the number of invasive cells in the OE-SCO2 group was higher than that in the OE-NC group(P<0.05).Conclusions Knockdown of SCO2 weakens the migration and infiltration ability of MDA-MB-231 cells,while overexpression of SCO2 promotes the migration and infiltration ability of breast cancer MDA-MB-231 cells.SCO2 is expected to be a regulatory target for the treatment of breast cancer metastasis.