摘要
目的用前列腺素E2刺激结肠癌细胞,观察和探讨可否引起CacyBP/SIP核转位。方法根据hCacyBP/SIP序列设计引物,PCR扩增、酶切、连接、转化等分子克隆法构建含有钙周期素结合蛋白(CacyBP/SIP)的慢病毒重组质粒,PCR鉴定、测序;将构建好的带有绿荧光蛋白的CacyBP-Lentivirus载体转染至结肠癌SW480细胞,用激光共聚焦显微镜检测CacyBP/SIP表达定位;前列腺素E2刺激后激光共聚焦显微镜、Western blot观察CacyBP/SIP在细胞中的定位。结果激光共聚焦和Westernblot发现:前列腺素E2刺激前CacyBP/SIP定位和表达主要在细胞胞质,用100μmol/L前列腺素E2刺激1h,CacyBP/SIP定位和表达在细胞胞质和胞核。结论前列腺素E2刺激可以引起CacyBP/SIP由胞质转位至细胞核。
Objective To observe and evaluate effect of Prostaglandin E2 on nuclear translocation of CacyBP/SIP by stimulating colorectal cancer with Prostaglandin E2. Methods To design the primer according to the sequenceh of CacyBP/SIP,and to build lentivirus recombinant plasmid contains CacyBP/SIP by cloning. To transfect the over expressed lentiviral vector with Green fluo- rescent protein CacyBP/SIP into 8W480 colon cancer cells, and to test the location and expression of the CacyBP/SIP with Laser Scanning Confocal Microscope and the Western blot. To test the location of the CacyBP/SIP after the stimulation of Prostaglandin E2 with Laser Scanning Confocal Microscope and the Western blot. Results CacyBP/SIP were located and expressed in the periplasm,and both in periplasm and nuclear after 1 hour stimulation of 100 μmol/L Prostaglandin E2 with Laser Scanning Confocal Microscope and Western blot. Conclusion The Prostaglandin E2 can transfer CacyBP/SIP to the nucleus.
出处
《重庆医学》
CAS
CSCD
北大核心
2012年第29期3020-3022,共3页
Chongqing medicine
基金
国家自然科学基金资助项目(81072040)
