Evaluation of the intracellular trafficking of siRNAs in A375 cells by confocal laser scanning microscopy

doi:10.5246/jcps.2016.12.096

Journal of Chinese Pharmaceutical Sciences ›› 2016, Vol. 25 ›› Issue (12): 859-868.DOI: 10.5246/jcps.2016.12.096

• Original articles • Next Articles

Evaluation of the intracellular trafficking of siRNAs in A375 cells by confocal laser scanning microscopy

Yiping Diao, Jing Sun, Mengyi Yang, Bo Xu, Lihe Zhang, Zhenjun Yang^*

State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China

Received:2016-05-13 Revised:2016-06-27 Online:2016-12-21 Published:2016-07-14
Contact: Tel.: +8601082802503, E-mail: yangzj@bjmu. edu.cn
Supported by:
Ministry of Science and Technology of China (Grant No. 2012AA022501, 2012CB720604), and the National Natural Science Foundation of China (Grant No. 20932001, 81302626).

Abstract

Abstract:

Investigation intracellular trafficking of siRNAs following their delivery to cells is of great interest to elucidate dynamics of siRNA in cytoplasm. In this study, we present a novel confocal laser scanning microscopy (CLSM) method to evaluate a novel delivery system of 3′-peptide-siRNA therapeutic, which was named 3′-pAs-siRNA/CLD. This method could not only calculate the content of the intracellular 3′-peptide-siRNA, but also quantify its co-localization with cellular substructure. We observed that 3′-pAs-siRNA/CLD, which provided the better antitumor capability, also had a better cell uptake, endosome escape and a longer retention time in A375. This novel strategy was proved to be efficient, quantified and visualized, thus making the dynamics research of siRNA in cytoplasm clear and simplified.

Key words: siRNA/CLD complex, Endosomal escape, Cytoplasmic distribution, Confocal laser scanning microscopy

CLC Number:

R916

Supporting:

Yiping Diao, Jing Sun, Mengyi Yang, Bo Xu, Lihe Zhang, Zhenjun Yang. Evaluation of the intracellular trafficking of siRNAs in A375 cells by confocal laser scanning microscopy[J]. Journal of Chinese Pharmaceutical Sciences, 2016, 25(12): 859-868.

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Evaluation of the intracellular trafficking of siRNAs in A375 cells by confocal laser scanning microscopy

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