ABSTRACT
<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>Ectopic expression of PC-1 gene in NIH3T3 cells can induce malignant transformation of mouse fibroblastic cells both in vitro and in vivo.</p>
Subject(s)
Animals , Mice , Cell Line, Transformed , Cell Transformation, Neoplastic , Gene Expression , Genes, Neoplasm , Physiology , Mice, Inbred BALB C , Mice, Nude , NIH 3T3 Cells , Neoplasm Proteins , Genetics , Physiology , RNA, Messenger , Genetics , Random Allocation , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To study the effect of PC-1 gene knockdown on the biological action of prostate cancer cell line C4-2.</p><p><b>METHODS</b>Recombinant plasmids of expressing short hairpin RNA targeting PC-1 mRNA were constructed using DNA recombinant technology and transfected into C4-2 cells via liposome. The positive cell clones were selected by G418. The expression of PC-1 gene was analyzed by RT-PCR and Western blotting technology. MTT and soft agar cloning formation were applied to observe the changes of the growth rate and independent anchor ability of C4-2 cells.</p><p><b>RESULTS</b>PC-1 RNA interference severely affected the expression of PC-1 gene and reduced the growth and colony formation ability of C4-2 cells.</p><p><b>CONCLUSION</b>RNA interference-mediated PC-1 gene knockdown can decrease the growth and cloning formation ability of C4-2 cells.</p>