tRF-Pro-CGG对小鼠胰腺癌细胞生物学行为的影响及其分子机制  

Effect of tRF-Pro-CGG on the biological behavior of mouse pancreatic cancer cells and its molecular mechanism

作　　者：符庆胜 金雷 张旭东 徐荧晨 朱春富[2] 秦锡虎 吴宝强 FU Qingsheng;JIN Lei;ZHANG Xudong;XU Yingchen;ZHU Chunfu;QIN Xihu;WU Baoqiang(School of Graduate,Bengbu Medical College,Bengbu 233030,Anhui Province,China;Department of Hepatobiliary and Pancreatic Surgery,Changzhou No.2 People’s Hospital,Changzhou 213003,Jiangsu Province,China)

机构地区：[1]蚌埠医学院研究生院,安徽蚌埠233030 [2]常州市第二人民医院肝胆胰外科,江苏常州213003

出　　处：《中国癌症杂志》2023年第3期241-249,共9页China Oncology

基　　金：常州市卫健委科技项目(ZD202023)。

摘　　要：背景与目的:tRNA衍生的片段(tRNA-derived fragments,tRF)是一类长度为14~30 nt的小分子非编码RNA,其影响着恶性肿瘤的发展进程。本研究旨在探讨tRF-Pro-CGG对小鼠胰腺癌细胞生物学行为的影响及其可能的分子机制。方法:采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)检测tRFPro-CGG在小鼠胰腺癌细胞系pan02、LTPA,人胰腺癌细胞系Capan-2和正常胰腺细胞HPDE6-C7中的表达水平。通过慢病毒转染技术过表达pan02细胞及敲低LTPA细胞中tRF-Pro-CGG的表达,采用RTFQ-PCR和蛋白质印迹法(Western blot)检测过表达和敲低效果。采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)检测细胞增殖情况。采用transwell实验检测细胞迁移和侵袭能力。采用动物模型检测tRF-Pro-CGG对胰腺癌裸鼠移植瘤生长和转移的影响。采用H-E染色观察移植瘤的组织病理学结构。采用Western blot检测移植瘤组织中Ki-67增殖指数、转移相关蛋白E-钙黏蛋白(E-cadherin)、波形蛋白(vimentin)的表达及磷脂酰肌醇3-激酶(phosphoinositide 3-kinase,PI3K)/蛋白激酶B(protein kinase B,AKT)信号转导通路蛋白的表达及磷酸化情况。结果:小鼠胰腺癌细胞系pan02中tRF-Pro-CGG表达最低,在pan02细胞中转染tRF-Pro-CGG mimics后,tRF-Pro-CGG的mRNA和蛋白表达水平均显著升高(P<0.01),细胞增殖能力显著降低(P<0.01),细胞迁移(P<0.001)和侵袭能力(P<0.001)显著降低,裸鼠移植瘤体积(P<0.01)和重量(P<0.001)均显著降低,裸鼠移植瘤组织中出现明显坏死和凋亡细胞;裸鼠移植瘤组织中Ki-67增殖指数和vimentin的表达显著降低(P<0.001),而E-cadherin的表达显著升高(P<0.001),PI3K、P-PI3K、AKT和P-AKT的表达显著降低(P<0.001),胰腺癌肝转移例数差异无统计学意义(P>0.05)。小鼠胰腺癌细胞系LTPA中tRF-Pro-CGG表达最高,在LTPA细胞中转染tRF-Pro-CGG inhibitor后,tRF-Pro-CGG的mRNA和蛋白表达水平显著�Background and purpose:tRNA-derived fragments(tRF)are a kind of short non-coding RNA(14-30 nt)that influences the course of cancer.This study aimed to investigate the molecular pathways that might underlie the effects of tRF-Pro-CGG on the biological behavior of mouse pancreatic cancer cells.Methods:Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)was used to assess the expression of tRF-Pro-CGG in mouse pancreatic cancer cell lines pan02 and LTPA,human pancreatic cancer cell line Capan-2,and normal pancreatic cells HPDE6-C7.tRF-Pro-CGG overexpression in pan02 cells and LTPA cell suppression were achieved through lentiviral transfection,and RTFQ-PCR and Western blot were used to determine overexpression and knockdown effects.Cell counting kit-8(CCK-8)was used to detect cell proliferation.Transwell assays were used to detect cell migration and invasion ability.The effect of tRF-Pro-CGG on the growth and metastasis of pancreatic cancer transplantation tumors in nude mice model was investigated.H-E staining was used to observe the histopathological structure of transplantation tumors.Western blot was used to detect the expression and phosphorylation of proliferation-related protein Ki-67 and metastasis-related proteins.Western blot was used to assess the expressions of cadherin,vimentin,phosphoinositide 3-kinase/protein kinase B(PI3K/AKT)pathway protein and phosphorylation in transplanted tumor tissues.Results:tRF-Pro-CGG expression was lowest in the mouse pancreatic cancer cell line pan02.Both mRNA and protein expression levels of tRFPro-CGG were significantly increased(P<0.01)after transfection of tRF-Pro-CGG mimics in pan02 cells,and cell proliferation ability(P<0.01),cell migration(P<0.001)and invasion ability(P<0.001)were significantly reduced.A significant decrease in the volume(P<0.01)and weight(P<0.001)of transplanted tumors in nude mice was observed,and significant necrotic and apoptotic cells in transplanted tumor were identified.In transplanted tumor tissues of nude mice,the Ki-67 pr

关 键 词：胰腺癌 tRNA衍生的片段-Pro-CGG 增殖 迁移 侵袭 裸鼠移植瘤模型 

分 类 号：R735.9[医药卫生—肿瘤]