抑制长链非编码RNA FAL1表达对上皮性卵巢癌细胞生物学行为的影响及其分子机制

摘要

Objective:To explore the difference in the expression of long chain non-coding RNA (lncRNA) FAL1 between epithelial ovarian cancer cells and normal human ovarian cells and the effect of lncRNA FAL1 expression in ovarian cancer cell line SKVO3 on invasion,migration and apoptosis of ovarian cancer cells.Methods:Using real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) technique to detect the FAL1 expression level of lncRNA in tissues;the design and synthesis of FAL1-siRNA primer sequence was transfected into SKVO3,detection of ovarian cancer cell migration by cell scratch test,Transwell to detect the invasive ability of ovarian cancer cells,detect the apoptosis of ovarian cancer cells by flow cytometry (Western blotting),Western blotting detection of phosphorylated extracellular signal regulated protein kinase 1/2 (p-ERK1/2) and phosphorylation of mitogen extracellular kinase 1/2 (p-MEK1/2) protein expression level.Results:The expressionoflncRNA FAL1 in epithelial ovarian cancer tissues was higher than that of normal ovarian tissue [(15.04±2.24) vs.(2.93 ±0.39),P ＜0.05].After silencing the expression of lncRNA FAL1,the apoptotic ability of ovarian cancer cells was significantly enhanced [(18.38±0.73)％ vs.(2.86±0.09)％,P＜0.05],while the migration and invasion ability of ovarian cancer cells were decreased [(6.68±1.49) μm vs.(12.85±2.56) tμm,(25.80±2.59) vs.(145.6±5.23),P＜0.05],and the phosphorylation level of MEK1/2 and ERK1/2 protein in MEK/ERK pathway was significantly decreased (P＜0.05).Conclusions:lncRNA FAL1 can influence the invasion,migration and apoptosis of epithelial ovarian cancer cells by activating MEK/ERK pathway,and its abnormal expression may be an important molecular mechanism of occurrence and development of ovarian cancer.%目的:探索上皮性卵巢癌细胞与正常人卵巢细胞中长链非编码RNA (lncRNA) FAL1的表达差异及沉默卵巢癌细胞株SKVO3中lncRNA FAL1的表达对卵巢癌细胞侵袭、迁移及凋亡的影响.方法:利用实时荧光定量聚合酶链反应(qRT-PCR)技术检测组织中lncRNA FAL1表达水平;设计并合成FAL1-siRNA引物序列转染SKVO3,通过细胞划痕试验检测卵巢癌细胞的迁移能力,Transwell侵袭实验检测卵巢癌细胞的侵袭能力,流式细胞术检测卵巢癌细胞的凋亡,蛋白质印迹(Western blotting)检测磷酸化细胞外信号调节蛋白激酶1/2(p-ERK1/2)和磷酸化丝裂原细胞外激酶1/2(p-MEK1/2)蛋白的表达水平.结果:lncRNA FAL1在卵巢癌组织中的表达高于正常卵巢组织[(15.04±2.24) vs.(2.93±0.39),P＜0.05].沉默lncRNA FAL1的表达后,卵巢癌细胞的凋亡能力显著增强[(18.38±0.73)％ vs.(2.86±0.09)％,P＜0.05],而卵巢癌细胞迁移、侵袭能力均降低[(6.68±1.49)μm vs.(12.85±2.56)μm,(25.80±2.59)个vs.(145.6±5.23)个,均P＜0.05],MEK/ERK通路MEK1/2和ERK1/2蛋白磷酸化水平明显降低(P＜0.05).结论:lncRNA FAL1通过激活MEK/ERK通路影响上皮性卵巢癌细胞的侵袭、迁移及凋亡,其表达异常增高可能是卵巢癌发生发展的重要分子机制.