Expression of vascular endothelial growth factor mRNA in GI-101A and HL-60 cell lines.
Vascular endothelial growth factor (VEGF) is a potent angiogenic factor that has a strong association with growth and metastasis of various cancers. We analyzed the expression of VEGF mRNA levels in human breast-tumor derived GI-101A cells and in human promyelocytic leukemia derived HL-60 cells using RT-PCR technique. During our RT-PCR analysis we detected the expression of three splice variants of VEGF mRNA at 400, 520 and, 650 bp lengths, which were amplified by a single set of VEGF specific forward and reverse primers. The three RT-PCR products detected by us in these cells correspond to the mRNA splice variants coding for the three isoforms of VEGF respectively, VEGF(121), VEGF(165), and VEGF(189). Treatment of GI-101A and HL-60 cells with phorbol 12, 13-dibutyrate (PDB) or diethylstilbestrol (DES) resulted in a significant increase of VEGF mRNA levels in a dose dependent manner. Both treatments increased the levels of all three splice variants of VEGF mRNA and a maximum increase was detected with 10 microM concentrations of PDB or DES treatments after 2 h. Interestingly, both PDB and DES mediated stimulation of VEGF mRNA expression was completely blocked by the PKC inhibitor chelerythrine. Quantitation of VEGF levels by ELISA technique confirmed that changes seen in mRNA levels following different treatments altered the release of VEGF. Our results suggest that PDB and DES mediated effects on VEGF expression in GI-101A and HL-60 cells occur at the gene transcription level.
Ramakrishnan
R
R
Department of Pharmaceutical Sciences, College of Pharmacy, Ft. Lauderdale, Florida 33328, USA.
Zell
J A
JA
Malavé
A
A
Rathinavelu
A
A
D000470:Alkaloids; D017398:Alternative Splicing; D000970:Antineoplastic Agents / Q000494:pharmacology; D053119:Benzophenanthridines; D001943:Breast Neoplasms; D004054:Diethylstilbestrol / Q000494:pharmacology; D016228:Endothelial Growth Factors / Q000235:genetics*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D014644:Genetic Variation; D018922:HL-60 Cells; D006801:Humans; D008222:Lymphokines / Q000235:genetics*; D010617:Phenanthridines / Q000494:pharmacology; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013629:Tamoxifen / Q000494:pharmacology; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D000470:Alkaloids; D000970:Antineoplastic Agents; D053119:Benzophenanthridines; D016228:Endothelial Growth Factors; D008222:Lymphokines; D010617:Phenanthridines; D012333:RNA, Messenger; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D013629:Tamoxifen; D015240:Phorbol 12,13-Dibutyrate; D004054:Diethylstilbestrol; C016299:chelerythrine