RNA结合基序蛋白15B对胰腺癌细胞增殖与侵袭功能的影响及其机制  

Effects of RBM15B on proliferation and invasion of pancreatic cancer cells and underlying mechanism

作　　者：谭淳中 夏鹏 郭德良[1] 杨张朔 刘朋朋[1] 张浩[1] 刘志苏[1] Tan Chunzhong;Xia Peng;Guo Deliang;Yang Zhangshuo;Liu Pengpeng;Zhang Hao;Liu Zhisu(Department of Hepatobiliary&Pancreatic Surgery,Zhongnan Hospital,Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉大学中南医院肝胆胰外科,430071

出　　处：《中华实验外科杂志》2022年第6期1110-1113,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金项目(81772926)。

摘　　要：目的观察RNA结合基序蛋白15B(RBM15B)在胰腺癌(PC)的表达并探讨其与胰腺癌细胞增殖、侵袭的关系。方法通过实时荧光定量聚合酶链反应(RT-qPCR)技术和蛋白质免疫印迹(Western blot)技术检测RBM15B在80例PC组织、癌旁组织、胰腺癌细胞系(SW1990、HPAFII)和人类正常胰腺细胞(HPDE)中的mRNA和蛋白表达量。实验组转染sh1-RBM15B、sh2-RBM15B, 对照组转染sh-NC, 获得稳定的敲低RBM15B和对照SW1990细胞系。细胞计数试剂盒(CCK-8)增殖实验、克隆形成实验观察敲低RBM1B的实验组与NC对照组细胞增殖能力;Transwell侵袭实验研究敲低RBM1B的实验组与NC对照组细胞侵袭能力;RNA-seq研究RBM15B涉及的下游信号通路。两组间比较采用独立样本t检验、χ^(2)检验。结果胰腺癌组织RBM15B的蛋白表达量显著高于癌旁组织(2.87±0.41比1.07±0.38, t=5.623, P<0.01)。RBM15B mRNA表达水平与胰腺癌患者的TNM分期(χ^(2)=4.363, P<0.05), 肿瘤直径(χ^(2)=6.626, P<0.05)和淋巴结转移(χ^(2)=4.949, P<0.05)显著相关。胰腺癌细胞系RBM15B蛋白水平显著高于正常胰腺细胞系(SW1990:3.34±0.44;HPAFII:2.76±0.32比HPDE:0.81±0.17, t=6.421、4.779, P<0.01), 差异有统计学意义。低表达组SW1990在72 h细胞吸光度值显著低于对照组SW1990的吸光度(1.02±0.11、1.35±0.13比1.96±0.14, t=4.007、3.748, P<0.01)。低表达组SW1990形成的克隆集落数显著少于对照组SW1990数量(64.57±4.77、134.26±10.32比193.74±19.58, t=4.766、3.595, P<0.05)。低表达组SW1990穿过基底膜的细胞数量显著少于对照组SW1990数量(36.48±3.73、47.68±4.29比105.32±13.23, t=7.471、4.617, P<0.05)。RNA-seq富集分析结果提示, 差异基因显著富集在Hippo信号通路;低表达组SW1990的Yes相关转录调控因子(YAP)表达水平显著低于对照组SW1990的YAP表达水平(1.78±0.12、1.12±0.09比2.45±0.21, t=5.458、6.897, P<0.01);低表达组SW1990的磷脂-溶血磷脂转酰基酶(TAZ)表达水平显著低于�Objective To observe the expression of RNA-binding motif protein 15B(RBM15B)in pancreatic cancer(PC)and explore its relationship with the proliferation,invasion of pancreatic cancer cells.Methods Real-time quantitative polymerase chain reaction(RT-qPCR)and Western blotting were used to detect the mRNA and protein expression of RBM15B in 80 cases of PC tissues,adjacent tissues,two pancreatic cancer cell lines(SW1990,HPAFII)and human normal pancreatic cells(HPDE).The experimental group was transfected with sh1-RBM15B and sh2-RBM15B,and the control group was transfected with sh-NC.After two weeks of puromycin selection,stable knockdown RBM15B and control cell lines were obtained.The cell counting kit-8(CCK-8)proliferation assay and clone formation assay were used to detect cell proliferation ability.Cell invasion ability was measured by Transwell invasion assay.The comparison between the two groups was performed by independent samples t test andχ^(2) test.Results The protein expression of RBM15B was significantly up-regulated in pancreatic cancer tissues as compared with adjacent tissues(2.87±0.41 vs.1.07±0.38,t=5.623,P<0.01).RBM15B mRNA expression level was significantly correlated with TNM stage(χ^(2)=4.363,P<0.05),tumor diameter(χ^(2)=6.626,P<0.05)and lymph node metastasis(χ^(2)=4.949,P<0.05)in PC patients.The protein level of RBM15B in PC cell line(SW1990:3.34±0.44;HPAFII:2.76±0.32)was significantly higher than that in normal pancreatic cell line(HPDE:0.81±0.17),and the difference was statistically significant(t=6.421,4.779,P<0.01).The absorbance of SW1990 cells in the low expression group at 72 h(1.02±0.11,1.35±0.13)was significantly lower than that of the control group(1.96±0.14,t=4.007,3.748,P<0.01).The number of colonies formed by SW1990 in the low expression group(64.57±4.77,134.26±10.32)was significantly less than that in the control group(193.74±19.58,t=4.766,3.595,P<0.05).The number of SW1990 cells passing through the basement membrane in the low expression group(36.48±3.73,47.68±4.29)wa

关 键 词：RNA结合基序蛋白15B 胰腺癌 增殖 侵袭 Hippo通路 

分 类 号：R735.9[医药卫生—肿瘤]