赖氨酸特异性去甲基化酶1增强胰腺癌PanC1细胞E3泛素连接酶对p16的抑制作用  

The effect of lysine - specific demethylase 1 on E3 ligase repressing p16 transcription in pancreatic cancer cells

作　　者：陈辉星[1] 陈实[3] 李小燕[2] 陈明源[1] 严茂林[3] 黄龙[3] 

机构地区：[1]福建医科大学附属协和医院肝胆外科,福州350001 [2]福建医科大学附属协和医院病理科,福州350001 [3]福建省立医院肝胆外科,福建医科大学附属省立临床医学医学院,福州350001

出　　处：《中华实验外科杂志》2017年第11期1922-1924,共3页Chinese Journal of Experimental Surgery

基　　金：福建省卫计委中青年骨干人才项目基金（2013-ZQN-JC-17）

摘　　要：目的观察胰腺癌PanC1细胞中组蛋白赖氨酸特异性去甲基化酶1（LSD1）对E3泛素连接酶（Ring1B）抑制抑癌基因p16转录及表达作用的影响。方法通过采用短发夹RNA（shRNA）分别在PanC1细胞中敲除Ring1B（重复2组为shRing1B#1、#2）和LSD1基因（shLSD1#1、#2），未敲除组为shLuc，分别通过实时定量反转录聚合酶链反应（RT-qPCR）和Western blot检测PanC1细胞LSD1、Ring1B和p16基因的表达；而后通过质粒转染过表达Ring1B，再敲除LSD1基因，检测PanC1细胞p16的表达。结果Ring1B基因敲除后RT-qPCR结果显示p16 mRNA转录增加，shRing1B#1、shRing1B#2（4.613±0.416、4.615±0.224）高于shLuc（1.010±0.032）（P＝0.000）；Western blot结果显示Ring1B基因敲除组p16蛋白表达增加；LSD1基因敲除后p16 mRNA转录和蛋白表达也增加，shLSD1#1、shLSD1#2组和shLuc组对应数值分别为4.704±0.503、3.885±0.011和1.008±0.027（P＝0.005）。Ring1B过表达的PanC1细胞中（Ring1B组），LSD1敲除组shLSD1＋vector组（5.893±0.986）高于对照组shLuc＋vector组（1.010±0.201）；shLSD1＋Ring1B组（0.695±0.031）高于shLuc＋Ring1B组（0.156±0.016）（P＝0.001）。结论LSD1与Ring1B协同作用，共同参与抑制胰腺癌p16的转录表达；LSD1能够增强Ring1B抑制p16转录的作用。Objective To investigate the effect of lysine - specific demethylase 1 （ LSD1 ） on E3 ligase （Ringl B ） repressing p16 transcription in pancreatic cancer cells. Methods LSD1 and Ringl B genes were knocked down by short hairpin RNA （shRNA） in pancreatic cancer cell line PanC1, then the expression of LSD1, RinglB and p16 genes in PanC1 cells was detected by Western blotting and Real - time PCR respectively. Finally, LSD1 was knocked down in RinglB -overexpressing PanC1 cells, and the expression of p16 was detected by Western blotting and real - time quantitative PCR. Results After RinglB gene knockout, real -time quantitative reverse transcriptase -poiymerase chain reaction （RT-qPCR） showed that p16 and mRNA transcription increased, shRinglB#1 and shRinglB#2 （4. 613±0. 416 and 4. 615 ±0. 224） were higher than shLuc （ 1. 010 ±0. 032） （P =0. 000） ; whereas the corresponding values of shLSD1 #1, shLSD1 #2 and shLuc were 4. 704 ± 0. 503, 3. 885 ± 0. 011 and 1. 008 ± 0. 027, respectively （ P = 0. 005 ）. The knockdown of LSD1 in Ringl B overexpressing PanC1 cells could attenuate the repression of RinglB on p16 gene transcription （ 1. 010 ± 0. 201 vs. 5. 893 ± 0. 986, and 0. 156 ± 0. 016 vs. 0. 695 ± 0. 031, P = 0.001 ）. Conclusion LSD1 enhanced the repression of RinglB on p16 gene transcription in pancreatic cancer cells.

关 键 词：胰腺癌 赖氨酸特异性去甲基化酶1 E3泛素连接酶 P16 

分 类 号：R735.9[医药卫生—肿瘤]