Synthesis nano-carrier SWCNT to deliver Bcl-2 siRNA and induction of U251 cell apoptosis
HU Xiao-fang1,2, QIU Ren-jie4, YU Lei2, LIU Mao-sheng1, YANG Lin1, WANG Guo-bao2,3
Chinese Journal of Clinical Anatomy ›› 2017, Vol. 35 ›› Issue (5) : 521-525.
Synthesis nano-carrier SWCNT to deliver Bcl-2 siRNA and induction of U251 cell apoptosis
Objective Design and synthesis Nano-Carrier of SWCNT-siRNA, and study the effect on glioma in vitro. Methods Design and synthesis Bcl-2 siRNA, and assemble them into nano-carrier. Characterization of the SWCNT-siRNA delivery system was measured by dynamic light scattering (DSL). U251 cell were cultured in vitro, and then treated with SWCNT-siRNA. The intracellular distribution of siRNA was detected by confocal laser scanning microscopy. The expression of Bcl-2 was detected by qPCR. Cells were stained with annexin V-FITC and propidium iodide (PI). The percentages of apoptotic and necrosis cells were quantified using a flow cytometer. Results Through particle size analysis, we proved that siRNA strongly adsorbed onto the surface of pristine SWCNTs to form the steady complex SWCNT-siRNA. siRNA labeled red fluorescence（cy3-siRNA）was traced by Laser Scanning Confocal Microscope and we found that siRNA was successful to be delivered into intracellular. The result of PCR showed that the expression of Bcl-2 was reduced in the group of SWCNT-siRNA. Flow Cytometer revealed SWCNT-siRNA could inhibit the growth of cells and induce early apoptosis and death. Conclusion SWCNT as nano-carrier could deliver Bcl-2 siRNA into intracellular, significantly cleaved the mRNA of Bcl-2, inducing lower levels of Bcl-2 expression and markedly triggering cell apoptosis.
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