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

作　　者：丁亚飞[1] 刘景明 刘若阳[1] 黄珍林 杨文龙 顾朝辉[1] 贾占奎[1] 杨锦建[1] Ding Yafei;Liu Jingming;Liu Ruoyang;Huang Zhenlin;Yang Wenlong;Gu Chaohui;Jia Zhankui;Yang Jinjian(Department of Urology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院泌尿外科,郑州450000

出　　处：《中华实验外科杂志》2024年第8期1772-1775,共4页Chinese Journal of Experimental Surgery

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;RM1:1406.00±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;RM1:704.20±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。ObjectiveTo 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 occurrence of epithelial-mesenchymal transition(EMT).MethodsThe cancer genome atlas(TCGA),Cioprotal 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.ResultsTCGA:Compared to normal prostate tissue,the expression of NOC2L gene was relatively higher in prostate cancer tissue(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).ConclusionNOC2L promotes cell proliferation and migration.During the occurrence of

关 键 词：前列腺癌 上皮-间充质转化 E盒结合锌指蛋白1 

分 类 号：R737.25[医药卫生—肿瘤]