癌基因NOC2L在上皮-间充质转化过程中抑制前列腺细胞迁移的机制
Mechanism of NOC2L protein suppressing the migration of prostate cells in epithelial-mesenchymal transition
丁亚飞 1刘景明 1刘若阳 1黄珍林 1杨文龙 1顾朝辉 1贾占奎 1杨锦建1
作者信息
- 1. 郑州大学第一附属医院泌尿外科,郑州 450000
- 折叠
摘要
目的 探究肿瘤促进基因NOC2L在上皮-间充质转化(EMT)过程中对前列腺癌细胞迁移的影响及分子机制.方法 采用数据库肿瘤基因组图谱(TCGA)、Cioprotal等分析NOC2L差异表达;Transwell、细胞计数试剂盒(CCK-8)、平板克隆等验证分子功能;蛋白质印迹法(Western blot)、聚合酶链反应(PCR)等探究分子机制.两组间比较采用独立样本t检验,多组之间的比较采用单因素方差分析.结果 TCGA:相较于正常前列腺组织,前列腺癌组织中NOC2L基因的表达量相对升高(前列腺癌组织与正常前列腺组织:5.83比5.43,P＞0.05);HPA:高级别前列腺癌中NOC2L含量更高(相对染色评分:高级别6.828±0.669比3.909±0.222,t=4.15,P＜0.05).敲低NOC2L抑制细胞迁移、增殖能力,敲低NOC2L后克隆形成数低于对照组(PC3:542.00±35.23比254.00±11.55,t=7.77,P＜0.05;RM1:1 406.00±15.34 比 754.00±75.92,t=8.41,P＜0.05);CCK-8 培养吸光度低于对照组(PC3:1.657±0.028 比 0.972±0.014,t=21.86,P＜0.05;RM1:1.643±0.007比 2.931±0.105,t=12.20,P＜0.05);细胞迁移明显低于对照组(PC3:704.00±26.12 比 152.80±6.08,t=20.55,P＜0.05;RM 1:704.20±37.56 比 208.40±7.26,t=12.96,P＜0.05).过表达 ZEB1在转录水平抑制 NOC2L 表达(PC3:1.001±0.028 比 0.377±0.018,t=18.62,P＜0.05;RM1:1.000±0.012比0.529±0.011,t=28.42,P＜0.05),NOC2L在ZEB1表达升高情况下功能逆转抑制细胞迁移,与ZEB1组比较,过表达NOC2L后迁移细胞减少(PC3:899.80±31.17比438.00±12.55,t=13.74,P＜0.05;RM1:986.80±65.83 比 466.40±9.67,t=7.82,P＜0.05).结论 NOC2L 促进细胞增殖及迁移,在EMT发生时NOC2L抑制CDH1功能减弱,通过抑制VIM和CDH2发挥抑制细胞迁移作用.
Abstract
Objective To investigate the effect of NOC2L on the migration of prostate cancer cells,and to systematically analyze the molecular mechanism of the contrary effect of NOC2L in the occur-rence of epithelial-mesenchymal transition(EMT).Methods The cancer genome atlas(TCGA),Ciopro-tal and HPA databases were used to analyze the differential expression of NOC2L.Transwell,cell counting kit-8(CCK-8)and plate cloning assays were used to verify the molecular functions.Western blotting and polymerase chain reaction(PCR)were used to explore the molecular mechanism.Results TCGA:Com-pared to normal prostate tissue,the expression of NOC2L gene was relatively higher in prostate cancer tis-sue(prostate cancer tissue vs.normal prostate tissue:5.83 vs.5.43,P＞0.05);HPA:NOC2L content was higher in high-grade prostate cancer(relative chromatin staining score:high-grade 6.828±0.669 vs.3.909±0.222,t=4.15,P＜0.05).NOC2L knockdown inhibited cell migration and proliferation,and the number of clones formed after NOC2L knockdown was less than that in control group(PC3:542.00±35.23 vs.254.00±11.55,t=7.77,P＜0.05;RM1:1406.00±15.34 vs.754.00±75.92,t=8.41,P＜0.05);The absorbance of CCK8 culture after NOC2L knockdown was lower than that of control group(PC3:1.657±0.028 vs.0.972±0.014,t=21.86,P＜0.05;RM1:1.643±0.007 vs.2.931±0.105,t=12.20,P＜0.05).Cell migration was significantly lower after NOC2L knockdown than that of control group(PC3:704.00±26.12 vs.152.80±6.08,t=20.55,P＜0.05;RM1:704.20±37.56 vs.208.40±7.26,t=12.96,P＜0.05).Overexpression of ZEB1 inhibited the mRNA expression of NOC2L(PC3:1.001±0.028 vs.0.377±0.018,t=18.62,P＜0.05;RM1:1.000±0.012 vs.0.529±0.011,t=28.42,P＜0.05),and NOC2L reversed function and inhibited cell migration when ZEB1 expression was elevated.Compared with ZEB1 group,after overexpression of NOC2L,the number of migrating cells decreased(PC3:899.80±31.17 vs.438.00±12.55,t=13.74,P＜0.05;RM1:986.80±65.83 vs.466.40±9.67,t=7.82,P＜0.05).Conclusion NOC2L promotes cell prolifera-tion and migration.During the occurrence of EMT,NOC2L inhibits the function of CDH1 and inhibits cell migration by inhibiting VIM and CDH2.
关键词
前列腺癌/上皮-间充质转化/E盒结合锌指蛋白1Key words
Prostate cancer/Epithelial-mesenchymal transition/Zinc finger E-box binding protein 1引用本文复制引用
出版年
2024
NETL
NSTL
万方数据