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| Transfection Efficiency Using PEI-CP Complex for CD133+ Differently Expressed by Colon Cancer Lines |
| DAI Shuang, ZHAO Qing-qing, QIU Feng |
| Department of Pharmaceutics, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China |
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Abstract Objective: To investigate transfect efficiency of CP/DNA at CSC marker CD133+ which is shown to be differently expressed in colon cancer cell lines, and to discuss the relationship between transfect and TR peptide, which is capable to specifically bind to CD133 by targeting CD133+ colon cancer cells. Methods: The expression of CD133+ in four cell lines were divided into 2 groups: the positive colon cell lines in SW480, HCT116, and the negative in CaCO2, SW620. Considering of PEI and Chitosan, they were linked together and composition,particle size,as well as the zeta potential of CP complex was measured.CCK-8 assay was also used to evaluate their toxicity. Using Chitosan-linked-PEI as the vector, PGL3 and EGFP plasmid transfect into four colon cancer cells after 48h incubation, and the CD133-related protein expressions were assessed by Western blotting analysis. Results: The CSC marker expression of CD133+ were significantly different.CD133+ was positive in more than 90% of colon cells, SW480 in 96.97%, HCT116 in 92.96%, however, low expression of CaCO2, SW620 was found in 2.00%, 0.56%.When the N/P of PEI/Chitosan/DNA was 5 ∶4 ∶1, CP complex shows a high transfect efficiency and low toxicity. And when the positive expression of CD133+ was added, the transfect efficiency was decreased. The result of Western blotting analysis showed that the ratios of CD133 in positive group were higher than those in negative group. Conclusion: The transfect efficiency of CP vector can increase the transfect efficient in low expression of CD133+.TR peptide-CD133 may facilitate CP vector into cells.
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Received: 17 February 2016
Published: 25 June 2016
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