Angiopoietin (Ang)-1 and Ang-2 interact in angiogenesis to activate the Tie-2
receptor, which may be involved in new vessel maturation and regression. Mast cells
(MCs) are also involved in formation of new blood vessels and angiogenesis. The
present study was designed to test whether MCs can mediate angiogenesis in myocardial
microvascular endothelial cells (MMVECs). Using a rat MMVEC and MC co-culture system,
we observed that Ang-1 protein levels were very low even though its mRNA levels were
increased by MCs. Interestingly, MCs were able to enhance migration, proliferation,
and capillary-like tube formation, which were associated with suppressed Ang-2
protein expression, but not Tie-2 expression levels. These MCs induced effects that
could be reversed by either tryptase inhibitor [N-tosyl-L-lysine
chloromethyl ketone (TLCK)] or chymase inhibitor
(N-tosyl-L-phenylalanyl chloromethyl ketone), with TLCK showing
greater effects. In conclusion, our data indicated that MCs can interrupt neovessel
maturation via suppression of the Ang-2/Tie-2 signaling pathway.
In the present study, we evaluated the effects of four solutions [Dulbecco's modified Eagle's medium (DMEM), sodium lactate Ringer's injection (SLRI), phosphate-buffered saline (PBS), and NaCl] on the transfection of the human protein kinase C-a antisense oligonucleotide (PKC-a ASO) aprinocarsen in human lung carcinoma A549 cells. Specifically, SLRI, DMEM, PBS, or NaCl were used as the growth solutions for A549 cells, and OPTI-MEM was used as the PKC-a ASO diluent for transfection. Additionally, SLRI, DMEM, PBS, or NaCl were used as both the growth solutions and diluents for transfection. The cell viability and transfection efficiency were determined. The results demonstrated that when SLRI was used as either the growth solution or both the growth solution and diluent for aprinocarsen transfection in A549 cells, the effects were close to the best effects observed with DMEM as the growth solution and OPTI-MEM as the diluent, which supported the transfection of aprinocarsen into the cells. Moreover, SLRI resulted in higher transfection efficiency than those of PBS and NaCl. In in vitro experiments, aprinocarsen effectively induced apoptosis in A549 cells. In conclusion, SLRI may replace PBS or NaCl in clinical trials as a transfection solution readily accepted by the human body. To our knowledge, this is the first report demonstrating the use of SLRI as a transfection solution in lung-cancer cell lines.
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