RegⅣ基因对胰腺癌细胞PANC-1增殖、侵袭、转移能力的影响  

Effects of RegⅣ on proliferation,invasion,and migration of pancreatic cancer cells PANC-1

作　　者：胡启立[1,2] 李健[2] 孟红波[2] 周波[2] 王锋[2] 宋振顺[1,2] 

机构地区：[1]安徽医科大学上海临床学院,上海200072 [2]上海市第十人民医院

出　　处：《山东医药》2017年第29期1-4,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81670582)

摘　　要：目的探讨RegⅣ基因对胰腺癌细胞PANC-1增殖、侵袭、转移能力的影响。方法取对数生长期的PANC-1细胞,于孵育箱中培养至70%~80%融合,分别以Puro-P-RegⅣ-shRNA慢病毒载体(P-RegⅣ-shRNA组)以及Puro-shRNA慢病毒空载体(P-NC-shRNA组)感染细胞,并设立不感染组(PANC-1组)。48 h后,分别用qRTPCR法和Western blotting法检测各组RegⅣmRNA及蛋白的表达。通过CCK-8试验检测细胞增殖情况,利用划痕试验和Transwell小室侵袭试验分别检测细胞的转移和侵袭能力。将10只5周龄的雌性裸鼠随机分成P-RegⅣ-shRNA裸鼠组和P-NC-shRNA裸鼠组,每组5只。取对数生长期的P-RegⅣ-shRNA和P-NC-shRNA组细胞分别注射入上述两组中。自接种日起每天观察肿瘤结节生长情况,6周后处死裸鼠并取出瘤体称重并测量瘤体体积。结果P-RegⅣ-shRNA组中RegⅣmRNA和蛋白的表达水平较PANC-1组和P-NC-shRNA组均下降(P均<0.05)。P-RegⅣ-shRNA组细胞增殖活力较P-NC-shRNA组和PANC-1组下降(P均<0.05)。与PANC-1组、P-NC-shRNA组比较,P-RegⅣ-shRNA组48 h细胞迁移距离缩短,穿膜细胞数减少(P均<0.05)。PANC-1组、P-NC-shRNA组各观察指标比较,P均>0.05。与P-NC-shRNA裸鼠组比较,P-RegⅣ-shRNA裸鼠组肿瘤生长速度慢,肿瘤体积小(P均<0.01)。结论下调RegⅣ基因可抑制胰腺癌细胞PANC-1增殖、转移、侵袭。Objective To explore the effects of RegⅣ gene on the proliferation,invasion,and migration of pancreatic cancer cells PANC-1. Methods PANC-1 cells in logarithmic growth phase were cultured in the incubator until they reached 70%-80% confluence. PANC-1 cells were transfected with Puro-P-RegⅣ-shRNA lentiviral vector（ P-RegⅣ-shRNA group） and Puro-shRNA lentiviral empty vector（ P-NC-shRNA group）,respectively. Cells without transfection or with null-transfection were served as controls（ PANC-1 group）. At 48 h after transfection,quantitative real-time reverse transcription-polymerase chain reaction（ qRT-PCR） and Western blotting were used to detect the expression of RegⅣ mRNA and protein. The CCK-8 test was used to detect the proliferation. Scratch test and Transwell chamber test were used to detect the migration and invasion of the cells,respectively. Ten 5-week-old female nude mice were randomly divided into the P-RegⅣ-shRNA nude mouse group and P-NC-shRNA nude mouse group with 5 in each group,which were then injected with P-RegⅣ-shRNA and P-NC-shRNA cells in logarithmic growth phase in the above two groups. The growth of tumor nodules was observed every day from the day of inoculation. After 6 weeks,the nude mice were sacrificed and the tumor volume was measured. Results The expression levels of RegⅣ mRNA and protein in the P-RegⅣ-shRNA group were significantly lower than those of the PANC-1 group and P-NC-shRNA group（ both P 0. 05）. Cells in the P-RegⅣ-shRNA group had a lower proliferative capacity than cells in the P-NC-shRNA group and PANC-1 group（ P 0. 05）. Moreover,compared with the PANC-1 group and P-NC-shRNA group,Puro-P-RegⅣ-shRNA group showed shorter transmembrane distance and decreased number of transmembrane cells,indicating decreased migration ability（ all P 0. 05）. No significant differences were found in the above parameters between the PANC-1 group and P-NC-shRNA group（ all P 0. 05）. Besides,tumor growth rate and the tumor volume in nude mice were dec

关 键 词：胰腺肿瘤 RegⅣ基因 再生基因家族 细胞增殖 细胞侵袭 

分 类 号：R737.9[医药卫生—肿瘤]