[Expression of estrogen receptor in different breast cancer cell lines transfected with estrogen receptor plasmid HEGO5 mediated by FuGENE6].
OBJECTIVE
Gene transfection is a major approach in studies of estrogen receptor (ER)expression in breast cancer cells,and ER's functional mechanism. This study was designed to detect the transfection efficiency of different human breast cancer cell lines mediated by FuGENE6 reagent, and ER expression in these cells after transfected with ER plasmid HEGO5.


METHODS
Breast cancer cell lines were transfected with plasmid pEGFP-N1,and the transfection efficiency was measured by flow cytometry (FCM). The cell lines with high transfection efficiency were transfected with HEGO5,and then the expression of ER was determined by FCM and Western blot.


RESULTS
There were obvious differences in transfection efficiencies of breast cancer cell lines MM-231, MM330, MM134 VI, MM175VII, MM157, MM361, MM436, MM453, UaCC812, UaCC893, BT-549, BT-20, HBL-100, Hs578t, MCF-7, T-47d, and ZR-75-1. Repetitious transfection test in HBL-100 cells showed that co-efficient of variation was 5.1. HEGO5 was transfected into the cells with high plasmid transfection efficiency (MCF-7, HBL-100, Hs578t, MM436, MM453, and BT-20), positive rates of ER in these cells ranged from 12.9% to 54.8%, corresponding to the pEGFP-N1 transfection efficiency, and the expression of ER detected by FCM were in accordance with those tested by Western blot. To some extent, the expression of ER in transfected cells was cell cycle specific.


CONCLUSIONS
Transfection of breast cancer cell lines mediated by FuGENE6 reagent has a good repetition,and the results were stable and reliable. In the cells transfected with HEGO5,the expression of ER can be qualitatively detected by Western blot, and quantitatively detected by FCM.
Molecular Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, P.R.China.
De-Ding
DD
Tao
Min
M
Jiang
Yong-Dong
YD
Feng
Jian-Hong
JH
Wu
Jian-Ping
JP
Gong
D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D002453:Cell Cycle; D045744:Cell Line, Tumor; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008055:Lipids; D010957:Plasmids; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
C411955:FuGene; D008055:Lipids; D011960:Receptors, Estrogen; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins