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浙江大学学报（医学版）
浙江大学学报（医学版）  2017, Vol. 46 Issue (4): 341-348    DOI: 10.3785/j.issn.1008-9292.2017.08.01
乳腺癌分子医学专题     
CD97免疫表位对乳腺癌细胞株MDA-MB231生物学行为的影响
田华, 陈洋, 赵建刚, 刘达人, 梁刚, 龚谓华, 陈力, 吴育连
浙江大学医学院附属第二医院外科, 浙江 杭州 310009
Effects of siRNAs targeting CD97 immune epitopes on biological behavior in breast cancer cell line MDA-MB231
TIAN Hua, CHEN Yang, ZHAO Jiangang, LIU Daren, LIANG Gang, GONG Weihua, CHEN Li, WU Yulian
Department of General Surgery, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China
 全文: PDF(5633 KB)  
摘要：

目的：明确CD97免疫表位CD97EGF和CD97Stalk对乳腺癌细胞生物学行为的影响。方法：采用siCatchTM siRNA design软件设计CD97EGF小干扰RNA（siRNA）和CD97Stalk siRNA并转染CD97高表达乳腺癌细胞株MDA-MB231；蛋白质印迹法筛选高敏感CD97EGFsiRNA和CD97Stalk siRNA。倒置显微镜观察和计数siRNA转染后乳腺癌细胞的生长；四唑盐（MTT）法检测siRNA转染后乳腺癌细胞的增殖活性；划痕试验和Transwell试验观察siRNA转染对乳腺癌细胞迁移和侵袭力的影响；TUNEL法检测siRNA转染对乳腺癌细胞凋亡的影响；流式细胞术检测siRNA转染对乳腺癌细胞周期的影响。结果：siRNA转染后，乳腺癌细胞的生长数量和增殖活性明显低于对照组，且CD97Stalk siRNA组的变化较CD97EGFsiRNA组更为显著（均P<0.05）；划痕试验结果显示，siRNA转染组乳腺癌细胞的移动速率明显慢于对照组（均P<0.01），且CD97Stalk siRNA对乳腺癌细胞移动速率的影响明显大于CD97EGFsiRNA（P<0.05）；Transwell试验结果显示，siRNA转染组迁移过膜的细胞数量明显少于对照组，且以CD97Stalk siRNA组减少最为显著（均P<0.05）。siRNA转染对乳腺癌细胞凋亡率的影响不明显，但siRNA转染组中G0/G1期细胞所占比例较对照组明显减小，而S期细胞所占的比例明显增加，且CD97Stalk siRNA对细胞周期的影响较CD97EGFsiRNA更为显著（均P<0.05）。结论：CD97蛋白分子可能参与乳腺癌细胞株的生长、增殖、迁移和侵袭等生物学行为，CD97Stalk免疫表位对于乳腺癌细胞株生物学行为影响较CD97EGF免疫表位更为显著。
关键词： RNA;  小分子干扰;  乳腺肿瘤/病理学;  抗原;  CD;  细胞凋亡;  细胞周期;  细胞增殖;  肿瘤浸润;  表位    
Abstract:

Objective:To investigate the effects of siRNAs targeting CD97 immune epitopes on proliferation, infiltration, apoptosis and cell cycle of breast cancer cells. Methods:siRNA sequences targeting CD97EGF and CD97Stalk immune epitopes were designed according to Gene Bank NM_001025160.2 with smart siCatchTM siRNA design software. CD97siRNAs were transfected into MDA-MB231 cells in which CD97 was highly expressed. Highest sensitive CD97EGF and CD97Stalk siRNA were screened by Western blotting. Inverted microscope was used to observe the growth of CD97siRNAs-transfected MDA-MB231 cells; the proliferation activity of MDA-MB231 cells was detected by MTT method; the wound healing assay and Transwell migration test were performed to examine the migration and infiltration ability of CD97EGF and CD97Stalk siRNA-transfected MDA-MB231 cells; the effects of CD97EGF siRNA and CD97Stalk siRNA on cell apoptosis and cell cycle of MDA-MB231 cells were detected by TUNEL and flow cytometry. Results:The growth and proliferation activity of CD97siRNAs-transfected MDA-MB231 cells were significantly lower than those in the control groups, and such differences were more significant in CD97Stalk siRNA-transfected group (all P<0.05); scratch test showed that the wound healing rate was lower in CD97siRNAs-transfected groups, especially in CD97Stalk siRNA-transfected group (all P<0.05); Transwell migration showed that the number of MDA-MB231 cells crossing through chambers were less in CD97siRNAs-transfected groups, especially in CD97Stalk siRNA-transfected group (all P<0.05); no significant difference in cell apoptosis was observed between CD97siRNAs-transfected groups and control groups; cell cycle detection showed that CD97siRNAs-transfected groups had less cells in G0/G1 phase and more cells in S phase compared with the control groups, and such effect on cell cycle was more marked in CD97Stalk siRNA-transfected group (all P<0.05). Conclusion:CD97 plays an important role in the cell growth, proliferation, migration and invasion of breast cancer MDA-MB231 cells, and compared with CD97EGF, CD97Stalk may have more effective inhibitory effects on cellular malignant behaviors.
Key words: RNA, small interfering    Breast neoplasms/pathology    Antigens, CD    Apoptosis    Cell cycle    Cell proliferation    Neoplasm invasiveness    Epitopes
收稿日期: 2017-04-10 出版日期: 2017-08-25
CLC:  R737.9  
基金资助:

浙江省自然科学基金（LY16H030007）
通讯作者: 陈力(1960-),男,博士,主任医师,教授,博士生导师,主要从事胃癌基础与临床研究;E-mail:chenli@mail.hz.zj.cn;http://orcid.org/0000-0003-3093-3402     E-mail: chenli@mail.hz.zj.cn
作者简介: 田华(1973-),男,博士研究生,副主任医师,主要从事CD97蛋白和肿瘤微环境细胞免疫研究;E-mail:zjuth@zju.edu.cn;http://orcid.org/0000-0002-7248-9699
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引用本文:

田华 等. CD97免疫表位对乳腺癌细胞株MDA-MB231生物学行为的影响[J]. 浙江大学学报（医学版）, 2017, 46(4): 341-348.

TIAN Hua, CHEN Yang, ZHAO Jiangang, LIU Daren, LIANG Gang, GONG Weihua, CHEN Li, WU Yulian. Effects of siRNAs targeting CD97 immune epitopes on biological behavior in breast cancer cell line MDA-MB231. Journal of ZheJiang University(Medical Science), 2017, 46(4): 341-348.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.1008-9292.2017.08.01    或    http://www.zjujournals.com/xueshu/med/CN/Y2017/V46/I4/341

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