沉默TRIM27对人黑色素瘤细胞C8161迁移、侵袭及上皮间质转化的抑制作用研究  

Study on the Inhibitory Effect of Silencing TRIM27 on Cell Migration,Invasion and Epithelial Mesenchymal Transformation in Human Melanoma C8161 Cells

作　　者：王敬茶 刘雪 邱思思 Wang Jingcha;Liu Xue;Qiu Sisi(Department of Dermatology,Hebei Petro Central Hospital,Langfang,Hebei 065000,China)

机构地区：[1]河北中石油中心医院皮肤科,河北廊坊065000

出　　处：《四川医学》2025年第2期151-156,共6页Sichuan Medical Journal

摘　　要：目的 探究沉默三结构域蛋白27(TRIM27)对人黑色素瘤C8161细胞活性、迁移、侵袭及上皮间质转化(EMT)的影响以及核因子κB(NF-κB)信号通路的调控作用。方法 体外培养C8161细胞,将C8161细胞分为对照组(Con,不做干预)、阴性对照组(si-NC,转染si-NC至C8161细胞)和沉默TRIM27组(si-TRIM27,转染si-TRIM27至C8161细胞),用实时荧光定量PCR(RT-qPCR)法检测鉴定各组细胞TRIM27表达水平。沉默TRIM27组细胞进一步分为si-TRIM27组、si-TRIM27+Y组(加入5μmol/L NF-κB通路抑制剂BAY11-7082)和si-TRIM27+A组(加入1μmol/L NF-κB通路激活剂Prostratin),干预24 h。细胞计数试剂盒-8(CCK-8)检测细胞活力,Transwell小室法检测5组细胞迁移和侵袭能力;Western Blotting法对EMT和NF-κB通路关键蛋白的表达进行测定。结果 si-TRIM27组的TRIM27表达水平显著低于si-NC组(P<0.05),证明si-TRIM27转染成功。与si-NC组相比,si-TRIM27组细胞活力、迁移、侵袭、N-cadherin、Vimentin、磷酸化(p)-NF-κB蛋白水平下降,E-cadherin蛋白水平升高,差异均有统计学意义(P<0.05);与si-TRIM27组相比,si-TRIM27+Y组中BAY11-7082增强了si-TRIM27对C8161细胞的上述作用(P<0.05),si-TRIM27+A组中Prostratin则削弱了si-TRIM27对C8161细胞的上述作用(P<0.05)。结论 沉默TRIM27可能通过下调NF-κB通路来抑制人黑色素瘤C8161细胞活力、迁移、侵袭及EMT进程。Objective To investigate the effect of silencing tridomain protein 27(TRIM27)on the activity,migration,invasion and epithelial-mesenchymal transition(EMT)of human melanoma C8161 cells and the regulatory effect of nuclear factor kappa B(NF-κB)signaling pathway.Methods C8161 cells were cultured in vitro and were divided into control group(Con,no intervention),negative control group(si-NC,transfected si-NC to C8161 cells),and TRIM27 silencing group(si-TRIM27,si-TRIM27 inhibitor was transfected into C8161 cells).TRIM27 in each groupwas detected by real-time fluorescent quantitative PCR(RT-qPCR).Silenced TRIM27 cells were further divided into si-TRIM27 group,si-TRIM27+Y group(adding 5μmol/L NF-κB pathway inhibitor BAY11-7082)and si-TRIM27+A group(adding 1μmol/L NF-κB pathway activator Prostratin),with intervention for 24 hours.Cell counting kit-8(CCK-8),Transwell chamber method was used to detect cell viability,cell migration and invasion ability in five groups.The expressions of key proteins in EMT and NF-κB pathway were determined by Western Blotting.Results TRIM27 in si-TRIM27 group was significantly lower than that in si-NC group(P<0.05),which proved that si-TRIM27 transfection was successful.Compared with si-NC group,cell viability,migration,invasion,N-cadherin,Vimentin,phosphorylated(p)-NF-κB protein levels in si-TRIM27 group were decreased,and E-cadherin protein levels were increased,all the differences were statistically significant(P<0.05).Compared with the si-TRIM27 group,BAY11-7082 in the si-TRIM27+Y group enhanced the above effects of si-TRIM27 on C8161 cells(P<0.05).Prostratin in the si-TRIM27+A group weakened the above effect of si-TRIM27 on C8161 cells(P<0.05).Conclusion Silencing TRIM27 may inhibit the viability,migration,invasion and EMT process of human melanoma C8161 cells by down-regulating the NF-κB pathway.

关 键 词：黑色素瘤 三结构域蛋白27 核因子ΚB信号通路 细胞活力 迁移 侵袭 上皮间质转化 

分 类 号：R739.5[医药卫生—肿瘤]