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Survivin antisense oligonucleotide mediated by polyamidoaminedendrimer liposome induces apoptosis of hepatic cancer cells
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Abstract:
Objective:To investigate the possibility of polyamidoaminedendrimer (PAMAM)-liposome for survivin antisense oligonucleotide (survivin-asODN) delivery system and explore the effects of PAMAM-liposome-survivin-asODN on survivin expression and apoptosis of human hepatic cancer cell line SMMC-7721. Methods: The liposome modified PAMAM (PAMAM-liposome) was synthesized with liposome and PAMAM. Survivin-asODN was combined with the PAMAM-liposome or PAMAM to form PAMAM-liposome-survivin-asODN or PAMAM-survivin-asODN complexes. The shape and size of the two complexes were observed under a transmission electron microscope and their zeta potentials were measured with a zeta analytical tool. The encapsulating efficiency and DNA loading level were determined by ultraviolet spectrophotometer using a centrifuging method. PAMAM-liposome-survivin-asODN and PAMAM-survivin-asODN were transfected into SMMC-7721 cells, and the transfection efficiency was measured. The protein expression of survivin in SMMC-7721 cells was measured by Western blotting, and the apoptosis of SMMC-7721 cells was assessed by flow cytometry. Results: PAMAM-liposome, PAMAM-liposome-survivin-asODN and PAMAM-survivin-asODN were successfully established. No significant difference appeared in diameter between PAMAM-liposome-survivin-asODN and PAMAM-survivin-asODN (\[189.33±15.42\] vs \[181.83±13.67\] nm, P>0.05), as well as the encapsulating efficiency and drug loading level, but the zeta potential of PAMAM-liposome-survivin-asODN was higher than that of PAMAM-survivin-asODN (\[42.83±7.14\] vs \[32.33±5.57\] mV, P<0.05). The transfection efficiency of PAMAM-liposome-survivin-asODN was higher than that of PAMAM-survivin-asODN (\[73.33±9.29)% vs \[60.67±7.81\]%, P<0.05) in SMMC-7721 cells. The expression of survivin protein in SMMC-7721 cells of PAMAM-liposome-survivin-asODN group was less than that of PAMAM-survivin-asODN group (24.67±11.74 vs 43.17±11.63, P<0.05), while the apoptosis rate was higher than that of PAMAM-survivin-asODN (\[73.31±12.59\]% vs \[52.67±12.19\]%, P<0.05). Conclusion: The PAMAM-liposome can delivery survivin-asODN into SMMC-7721 cells effectively and induce SMMC-7721 cell apoptosis.
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Project Supported:
Project supported by the Natural Science Foundation of Guangdong Province (No. 9151051501000071)
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