摘要
目的小动物活体成像直观观察转录因子Ascl2对结肠癌HT-29细胞裸鼠移植瘤生长的影响。方法构建Ascl2的shRNA干扰质粒,对人结肠癌HT-29细胞进行瞬时转染48 h,G418筛选、克隆挑取、扩增培养建立稳定转染的细胞系,采用小动物活体成像仪直观观察Ascl2-RNAi对裸鼠移植瘤生长的影响。结果成功构建Ascl2的shRNA干扰质粒,并进行测序验证;质粒转染HT-29细胞,经G418筛选后挑选稳定转染的克隆,进行扩增培养,成功建立稳定转染的细胞系,命名为shRNA-Ascl2/HT-29和shRNA-control/HT-29细胞,并采用Western-blot证实干扰效果(P<0.01);采用小动物活体成像仪直观观察发现Ascl2-RNAi抑制HT-29细胞裸鼠移植瘤的生长。结论 Ascl2-RNAi导致结肠癌HT-29细胞体外致瘤能力下降,小动物活体成像技术直观、灵敏度高、实验成本低,值得进一步推广应用。
Objective To observe the tumor growth arrest in vivo by Achaete scute -like 2（Ascl2） RNA inter- ference in colon cancer HT - 29 cells though Living Animal Imaging. Methods ShRNA - control and shRNA - Ascl2 vectors were constructed. Colon cancer HT- 29 cells were transfected for 48 h with the two vectors,then the transfected cells were selected with G418 and stably transfected cell lines were established. The tumor growth arrest in vivo by Ascl2 RNA interference in colon cancer HT -：29 cells was observed directly though Living Animal Imaging. Results ShRNA - control and shRNA - Ascl2 vectors were constructed successfully, and verified by Sequencing. The two stably transfected cell lines were named as shRNA -Ascl2/HT- ：29 and shRNA - control/HT -29 cells. Ascl2 RNA interference effects were verified by western - blot （P 〈 0.01 ）. The tumor growth arrest in vivo by Ascl2 RNA interference in colon cancer HT -29 cells was observed directly though Living Animal Imaging. Conclusion Ascl2 RNA interference leads to tumor growth arrest in vivo. Liv- ing Animal Imaging has the advantages of intuition, high sensitivity and low cost, so it is worthy of further ap- plication.
出处
《合肥医学院学报》
2014年第1期81-85,共5页
Journal of Zunyi Medical University
基金
国家自然科学基金项目(NO:81201685)
