[Malignant transformation of NIH3T3 cells induced by ectopic expression of PC-1 gene].
OBJECTIVE
To establish a mouse fibroblastic cell line stably transfected with PC-1 gene, and using such cell line to investigate tumor development and progression imposed by the ectopic expression of PC-1 gene.


METHODS
Eukaryotic expression vector pcDNA3.1(-)/myc-his-pc-1 was transfected into mouse fibroblast cell line NIH3T3 by lipofectamine. Stable transfectants were selected by G418. The integration and expression of ectopic PC-1 gene were analyzed by PCR and RT-PCR. Cytomorphological analysis, MTT, soft agar colony formation and nude mice tumorigenesis assay were used to evaluate the effects of PC-1 gene expression on tumor development and progression.


RESULTS
NIH 3T3 cell lines stably expressing PC-1 gene were successfully established and confirmed by PCR and RT-PCR analyses of the integration and expression of ectopic PC-1 gene. Comparing with the parental cell line and cells transfected with control vector, the PC-1 gene transfectants acquired several phenotypes of transformed cells: increasing growth rate, ability to grow and form cell colonies on soft agar, and becoming tumorigenic in nude mice.


CONCLUSIONS
Ectopic expression of PC-1 gene in NIH3T3 cells can induce malignant transformation of mouse fibroblastic cells both in vitro and in vivo.
Chang
Xiao-tong
XT
Institute of Biotechnology, Academy of Military Medical Science, Beijing 100850, China.
Liang
Rui-xia
RX
Zhou
Jian-guang
JG
Zhang
Hao
H
Li
Jie-zhi
JZ
Wang
Jian
J
Zhang
Hui
H
Huang
Cui-fen
CF
D000818:Animals; D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic; D015870:Gene Expression; D052138:Genes, Neoplasm / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D041681:NIH 3T3 Cells; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011897:Random Allocation; D014162:Transfection
D009363:Neoplasm Proteins; D012333:RNA, Messenger