摘要
目的:探究circ-RBM33能否靶向miR-383-3p/CKS1B轴抑制鼻咽癌细胞的增殖和侵袭。方法:qRT-PCR检测CNE1和HNEpC细胞中circ-RBM33、miR-383-3p表达。将CNE1细胞随机分为si-NC组、si-RBM33组、miR-NC组、miR-383-3p组和si-RBM33+anti-miR-383-3p组。分别检测细胞增殖(CCK-8法)、侵袭(Transwell法)和迁移(划痕实验)及细胞中CKS1B蛋白表达(蛋白质印迹法);荧光素酶活性实验检测circ-RBM33和miR-383-3p/CKS1B的靶向关系。结果:人鼻咽癌细胞CNE1中circ-RBM33相对表达量高于人鼻黏膜上皮细胞HNEpC,miR-383-3p相对表达量低于人鼻黏膜上皮细胞HNEpC(P<0.05)。si-RBM33组CNE1细胞中circ-RBM3相对表达量、细胞存活率、细胞侵袭数目、细划痕愈合率及细胞中CKS1B蛋白表达均低于si-NC组(P<0.05);si-RBM33+anti-miR-383-3p组CNE1细胞存活率、细胞侵袭数目、细胞划痕愈合率及细胞中CKS1B蛋白表达均高于si-RBM33组(P<0.05)。miR-383-3p组CNE1细胞中miR-383-3p相对表达量明显增加,细胞存活率、细胞侵袭数目、细划痕愈合率及细胞中CKS1B蛋白表达均低于miR-NC组(P<0.05)。转染WT-circ-RBM33/CKS1B时miR-383-3p组荧光素酶活性明显低于miR-NC组(P<0.0001)。结论:敲减circ-RBM33可能通过上调miR-383-3p表达,进而抑制CKS1B表达来抑制鼻咽癌细胞的增殖、侵袭和迁移。
Objective:To investigate whether circ-RBM33 can target the miR-383-3p/CKS1B axis to inhibit the proliferation and invasion of nasopharyngeal carcinoma cells.Methods:qRT-PCR was used to detect the expression of circ-RBM33 and miR-383-3p in CNE1 and HNEpC cells.CNE1 cells were randomly divided into si-NC group,si-RBM33 group,miR-NC group,miR-383-3p group,and si-RBM33+anti-miR-383-3p group.Cell proliferation(CCK-8 assay),invasion(Transwell assay),migration(scratch assay),and expression of CKS1B protein in cells(Western blot)were detected.Luciferase activity assay was performed to determine the targeting relationship between circ-RBM33 and miR-383-3p/CKS1B.Results:The relative expression of circ-RBM33 in human nasopharyngeal carcinoma CNE1 cells was higher than that in human nasopharyngeal mucosal epithelial HNEpC cells,while the relative expression of miR-383-3p was lower than that in HNEpC cells(P<0.05).In the si-RBM33 group,the relative expression of circ-RBM33 in CNE1 cells,cell viability,number of invading cells,scratch closure rate,and expression of CKS1B protein in cells were lower than those in the si-NC group(P<0.05).In the si-RBM33+anti-miR-383-3p group,the cell viability,number of invading cells,scratch closure rate,and expression of CKS1B protein in cells were higher than those in the si-RBM33 group(P<0.05).In the miR-383-3p group,the relative expression of miR-383-3p in CNE1 cells was significantly increased,while cell viability,number of invading cells,scratch closure rate,and expression of CKS1B protein in cells were lower than those in the miR-NC group(P<0.05).Transfection with WT-circ-RBM33/CKS1B resulted in significantly lower luciferase activity in the miR-383-3p group than in the miR-NC group(P<0.0001).Conclusion:Knockdown of circ-RBM33 may inhibit the proliferation,invasion,and migration of nasopharyngeal carcinoma cells by upregulating miR-383-3p expression and subsequently suppressing CKS1B expression.
作者
周广全
孙相波
卢海波
赵松
ZHOU Guangquan;SUN Xiangbo;LU Haibo(The First People's Hospital of Yancheng City,Jiangsu Yancheng 224001,China)
出处
《河北医学》
CAS
2024年第6期881-886,共6页
Hebei Medicine
基金
江苏省卫生健康委员会课题项目,(编号:ZD20210048)。
