敲低PLK1对脉络膜黑色素瘤细胞增殖的抑制作用及其机制  

Inhibitory effect of PLK1 knockdown on the proliferation of choroidal melanoma cells and its mechanism

作　　者：赵芃芃 白小芳 卢凤丽 李思园 谭丛 蒋胜群 郁佳[1] 秦梅[1] ZHAO Pengpeng;BAI Xiaofang;LU Fengli;LI Siyuan;TAN Cong;JIANG Shengqun;YU Jia;QIN Mei(Department of Ophthalmology,First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠医学院第一附属医院眼科,蚌埠233000

出　　处：《山西医科大学学报》2021年第9期1121-1127,共7页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81903142);安徽省高校自然科学基金重点项目(KJ2020A0590)。

摘　　要：目的探究敲低PLK1对脉络膜黑色素瘤C918细胞增殖的影响及其可能的分子机制。方法RT-qPCR和Western blot实验检测人脉络膜血管内皮细胞和脉络膜黑色素瘤细胞中PLK1表达。将C918细胞设置为阴性对照组(NC组)和两个PLK1敲低组(sh1组、sh2组)。CCK-8和克隆形成实验检测敲低PLK1对C918细胞增殖的影响。流式细胞仪检测PLK1敲低对细胞周期的影响。Western blot检测PLK1敲低对组蛋白H3T3ph修饰的影响。ChIP实验检测组蛋白H3T3ph修饰在靶基因启动子上的富集情况。结果与脉络膜内皮细胞相比,PLK1在脉络膜黑色素瘤C918细胞中显著高表达(P<0.05)。与NC组细胞比较,敲低组PLK1的mRNA和蛋白水平均降低(P<0.05),PLK1敲低组细胞增殖显著被抑制(P<0.05),敲低PLK1组细胞周期阻滞于S期(P<0.05)。相比于NC,敲低PLK1能抑制BUBR1基因的表达并降低组蛋白H3T3ph修饰水平。与IgG比较,H3T3ph修饰能够富集在BUBR1基因的启动子区域(P<0.05)。结论敲低PLK1能够抑制脉络膜黑色素瘤细胞增殖,其机制可能与PLK1通过介导H3T3ph修饰在BUBR1基因启动子上的富集进而促进BUBR1的基因转录有关。Objective To explore the effect of knocking down PLK1 on the proliferation of choroidal melanoma C918 cells and its possible molecular mechanism.Methods The expression of PLK1 in human choroidal vascular endothelial cells and choroidal melanoma cells was detected by RT-qPCR and Western blot.The C918 cells were divided into negative control group(NC group)and two PLK1 knockdown groups(sh1 group,sh2 group).The effect of PLK1 knockdown on the proliferation of C918 cells was tested using CCK-8 and clone formation experiments,and then the change of cell cycle was detected using flow cytometry.The effect of PLK1 knockdown on histone H3T3ph modification was detected by Western blot,and the enrichment of histone H3T3ph modification on the target gene promoter was tested by ChIP experiment.Results Compared with choroidal endothelial cells,PLK1 was significantly highly expressed in choroidal melanoma C918 cells(P<0.05).Compared with NC group,the mRNA and protein levels of PLK1 were reduced in knockdown groups(P<0.05),the cell proliferation was significantly decreased(P<0.05),and the cell cycle was arrested in the S phase(P<0.05).Compared with NC,the knockdown of PLK1 attenuated the expression of BUBR1 gene and reduced the level of histone H3T3ph modification.Compared with IgG,H3T3ph modification was enriched in the promoter region of BUBR1 gene(P<0.05).Conclusion Knockdown of PLK1 may inhibit the proliferation of choroidal melanoma cells by mediating the enrichment of H3T3ph modification on the BUBR1 gene promoter,thereby promoting BUBR1 gene transcription.

关 键 词：脉络膜黑色素瘤 PLK1 细胞增殖 BUBR1 

分 类 号：R739.7[医药卫生—肿瘤]