西安交通大学学报（医学版）Issue(6)：749-752,757,5.DOI:10.7652/jdyxb201506007
HtrA1表达被沉默的滋养细胞株HPT-8/pSilencer4.1-HtrA1的构建
Construction of trophoblastic cell line HPT-8/pSilencer4.1-HtrA1 with silenced expression of HtrA1
摘要
Abstract
Objective To construct the plasmid pSilencer4.1/HtrA1 and stably transfect human trophoblastic cell line HPT-8.Methods We detected the expression of HtrA1 in cell line HPT-8 with immunohistochemical SABC,constructed the plasmid pSilencer4.1/HtrA1,transfected human trophoblastic cell line HPT-8 with plasmid pSilencer4.1/HtrA1 and negative plasmid pSilencer4.1, and observed the cell expression after transfection. Results About 90% of cellular cytoplasm of HPT-8 was dyed brown while the nucleus was negatively stained. Selective concentration of G418 was 200 μg/mL for HPT-8 to be transfected pSilencer4.1/HtrA1.With RT-PCR and Western blot methods,the expression of HPT-8 transfected plasmid Psilence4.1/HtrA1 was significantly downregulated compared with that of cells with negative plasmid and normal cells (P < 0.05 ).There was no significant difference in absorbance value between blank plasmid group and HPT-8 group (P >0.05).Conclusion We successfully constructed stable trophoblastic cell line HPT-8/ pSilencer4.1-HtrA1 with silenced expression of HtrA1,which lays foundation for further research.关键词
滋养细胞/HPT-8/HtrA1/质粒Key words
trophoblastic cell/HPT-8/HtrA1/plasmid分类
医药卫生引用本文复制引用
宗璐,苟文丽,郭玉琳..HtrA1表达被沉默的滋养细胞株HPT-8/pSilencer4.1-HtrA1的构建[J].西安交通大学学报（医学版）,2015,(6):749-752,757,5.基金项目
陕西省自然科学基础研究项目(No.2014JQ2-8047),中央高校基本科研业务费专项资金资助(No.XJJ2014072) Supported by the Natural Science Basic Research of Shaanxi Province(No.2014JQ2-8047)and the Fundamental Research Funds for the Central Universities (No.XJJ2014072) （No.2014JQ2-8047）
