摘要
目的建立肿瘤细胞迁移过程的多角度、实时动态、定量评估方法。方法建立细胞迁移观测模型,分别加入小檗碱及Rg3培养24h,用活细胞工作站随机选取8个不同观测点,连续观测24h。所取得的数据从迁移面积、距离、单个细胞迁移速度、单位面积细胞增殖数量等多角度进行分析。结果该方法可以长时间观察划痕内细胞的迁移过程;小檗碱高、中剂量组(25,12.5μmol·L-1)可抑制A549细胞的迁移过程(P<0.01),且迁移面积、距离、速度及细胞增殖数量4项指标的结果基本一致,人参皂苷Rg3(0.1mmol·L-1)可以在各时间点(6,12,24h)抑制细胞迁移速度(P<0.01),24h可抑制细胞迁移面积及距离(P<0.01),但可促进细胞增殖(P<0.01)。结论该方法灵敏、简便、快速,可广泛应用于细胞迁移过程的动态观察。
Aim To establish a new,multi-parameter,real time and qualitative cell migration evaluation method. Methods Lung cancer cell line A549 was cultured on the glass bottom dish. After treated with different dosages of berberine or Rg3 for 24 hours,several scratching lines at the same dimension were made and observed by Living Cell Working Station. 8 observation areas were selected randomly and imaged continuously for 24 hours. Transferred Area（TA）,Transferred Distance（TD）,Single Cell Transferred Speed（SCTS）and the Cell Division Number among defined area（ CDN） were analyzed after getting sequence images. Results The focus stage and the incubation system were sufficient to keep cell proliferation and made it possible for long term observation. Berberine at 25 μmol·L^-1 and 12. 5 μmol·L^-1 could inhibit the migration of A549（P〈0.01）. The analysis results of TA,TD,SCTS and CDN were basically coincident. Rg3 at 0.1 mmol·L^-1 could inhibit SCTS and promote CDN in 6,12 and 24 h（P〈0.01）,while decrease both TA and TD in 24 hs. Conclusion The method is sensitive,rapid and simple to be applied in the research of tumor metastasis,wound healing and inflammatory response with real time,in-situ and multi-parameters.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2010年第1期128-131,共4页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No30600842)
中国财政部自主选题资助项目(Nozx-zzxt-2006-005)
关键词
细胞迁移
活细胞工作站
细胞划痕
动态观察
小檗碱
RG3
cell migration living cell working station cell scratch dynamic observation berberine Rg3
