[The establishment and identification of the stable transfectant of HeLa cell expressing human new gene (PCIA1)].
OBJECTIVE
To construct a stable transfectant of human cervical cancer cell line HeLa that can highly express human novel gene (PCIA1).


METHODS
RT-PCR technique was applied to amplify the cDNA of PCIA1 gene from total RNA isolated from HUVEC. After sequenced, the open reading frame (ORF) of PCIA1 cDNA was cloned into eukaryotic expression vector pcDNA3.1 (+) to form the recombinant plasmid named as pcDNA3.1-PCIA1. Then lipofectamine 2000 was used to transfected the pcDNA3.1-PCIA1 into HeLa cells. The PCIA1-transfected HeLa cells were selected with G418, for 4 weeks. Finally, RT-PCR and Western blot were used to detect the expression of human PCIA1 in HeLa cells.


RESULTS
By RT-PCR and Western blot, we found that the human novel gene PCIA1 stably expressed in HeLa cells.


CONCLUSIONS
This study indicated that human PCIA1 could be stably expressed in HeLa cells and its functions on tumor cells could be further analyzed by this stable transfectant of PCIA1.
Wang
Rui
R
State Key Laboratory of Biotherapy, Cancer Center, West China Hospital, Sichuan University, Chengdu, 610041 China.
Deng
Hong-xin
HX
Wang
Guo-qing
GQ
Yan
Fei
F
Peng
Feng
F
D000818:Animals; D000951:Antigens, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D002478:Cells, Cultured; D002888:Chromosomes, Human, Pair 19 / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D004730:Endothelium, Vascular / Q000166:cytology*; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D016366:Open Reading Frames / Q000235:genetics; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology
D000951:Antigens, Neoplasm; D018076:DNA, Complementary; D011994:Recombinant Proteins