摘要 目的观察lncRNA FAL1在卵巢癌细胞及其耐药细胞株中的表达差异,探索下调lncRNA FAL1对细胞化疗耐药性的影响及机制。方法qRT-PCR法检测SKOV3和COC1细胞及其耐药细胞株FAL1基因表达水平,转染FAL1-siRNA下调FAL1基因,MTT检测细胞增殖能力,Transwell法检测细胞侵袭能力,平板克隆形成实验检测细胞克隆能力,Western blot法检测细胞耐药相关蛋白MDR-1、MPR-1、ABCG2和MAPK通路相关蛋白p38 MAPK、ERK1/2、JNK磷酸化水平。将已转染FAL1-siRNA的SKOV3/DDP和COC1/DDP细胞注射到BALB/c裸鼠皮下,定期测量瘤体积并称重。结果与SKOV3和COC1细胞比较,SKOV3/DDP和COC1/DDP细胞对DDP的敏感度降低,FAL1基因的表达水平升高(P<0.01)。转染FAL1-siRNA后,SKOV3/DDP和COC1/DDP细胞对DDP的敏感度增加(P<0.01),侵袭能力(P<0.05)和克隆能力降低(P<0.01),细胞中MDR-1、MPR-1、ABCG2表达水平(P<0.01)和p38 MAPK、ERK1/2、JNK磷酸化水平降低(P<0.05),皮下移植瘤的体积和瘤质量显著降低(P<0.01)。结论下调lncRNA FAL1可显著降低卵巢癌耐顺铂细胞株的化疗耐药性,抑制耐药细胞在体内的增殖能力,其作用机制与抑制MAPK信号通路的活化有关。 Objective To observe the expression difference of lncRNA FAL1 in ovarian cancer cells and their drug-resistant cell lines,and to explore the effect and mechanism of lncRNA FAL1 down-regulation on cell chemotherapy resistance.Methods The expression levels of fal1 gene in SKOV3 and COC1 cells and their drug-resistant cell lines were detected by qRT-PCR.fal1 siRNA was transfected to downregulate fal1 gene expression.MTT was used to detect cell proliferation.Transwell method was used to detect cell invasion ability.Plate clone formation test was used to detect cell clone ability,and Western blot was used to detect MDR-1,mpr-1,ABCG2 and phosphorylation levels of p38 MAPK,ERK1/2 and JNK.SKOV3/DDP and COC1/DDP cells transfected with FAL1-siRNA were injected subcutaneously into BALB/c nude mice.The volume and mass of subcutaneous transplanted tumors were measured.Results Compared with SKOV3 and COC1 cells,SKOV3/DDP and COC1/DDP cells were less sensitive to DDP,and the expression levels of FAL1 gene increased(P<0.01).After transfection with FAL1-siRNA,the sensitivity of SKOV3/DDP and COC1/DDP cells to DDP increased(P<0.01),and the invasion(P<0.05)and cloning ability(P<0.01)decreased.The expression levels of MDR-1,MPR-1,ABCG2(P<0.01)and the phosphorylation levels of p38 MAPK,ERK1/2 and JNK(P<0.05)decreased.The volume and mass of subcutaneous transplanted tumors were significantly reduced(P<0.01).Conclusion Down-regulation of lncRNA FAL1 could significantly reduce the chemotherapy resistance of cisplatin-resistant ovarian cancer cell lines and inhibit the proliferation of drug-resistant cells in vivo.Its mechanism is related to inhibiting the activation of MAPK signaling pathway.
机构地区 郑州大学附属郑州中心医院妇产科
出处 《肿瘤防治研究》 CAS CSCD 2021年第4期333-340,共8页 Cancer Research on Prevention and Treatment
关键词 卵巢癌 化疗耐药性 lncRNA FAL1 MAPK信号通路 SKOV3/DDP细胞 Ovarian cancer Chemotherapy resistance LncRNA FAL1 MAPK signaling pathway SKOV3/DDP cells
