Chinese Journal Of Clinical Anatomy ›› 2017, Vol. 35 ›› Issue (5): 521-525.doi: 10.13418/j.issn.1001-165x.2017.05.009

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Synthesis nano-carrier SWCNT to deliver Bcl-2 siRNA and induction of U251 cell apoptosis

HU Xiao-fang 1,2, QIU Ren-jie 4, YU Lei 2, LIU Mao-sheng 1, YANG Lin1,  WANG Guo-bao 2,3   

  1. 1.Department of Anatomy, Zhuhai Campus of Zunyi Medical College, Guangdong Zhuhai 519041,China; 2. Department of Key Laboratory of Construction and Detection of Guangdong Province, Southern Medical University, Guangzhou 510515,China; 3. Department of Nephrology, Nanfang Hospital, Southern Medical University, Guangzhou 510515,China; 4.College of Jiangxi University of Traditional Chinese Medicine, Jiangxi Nanchang 330004, China
  • Received:2017-06-28 Online:2017-09-25 Published:2017-10-30

Abstract:

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.

Key words:  , Glioma; siRNA; SWCNT; Bcl-2; U251