Calcium based siRNA coating: a novel approach for knockdown of HER2 gene in MCF-7 cells using gold nanoparticles

Abdul Sami; S.M. Saqalan Naqvi; Mazhar Qayyum; Abida Raza Rao; Uteuliyev Yerzhan Sabitaliyevich; M. Sheeraz Ahmad

doi:10.14715/cmb/2020.66.6.19

Issue

Vol. 66 No. 6: Issue 6

Issue Published : October 6, 2020

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Calcium based siRNA coating: a novel approach for knockdown of HER2 gene in MCF-7 cells using gold nanoparticles

https://doi.org/10.14715/cmb/2020.66.6.19

Abdul Sami

University Institute of Biochemistry and Biotechnology, Pir Mehr Ali Shah, Arid Agriculture University, Rawalpindi, Pakistan

S.M. Saqalan Naqvi

University Institute of Biochemistry and Biotechnology, Pir Mehr Ali Shah, Arid Agriculture University, Rawalpindi, Pakistan

Mazhar Qayyum

Department of Zoology, Pir Mehr Ali Shah, Arid Agriculture University, Rawalpindi, Pakistan

Abida Raza Rao

Nanotheranostic Lab, National Institute of Lasers and Optronics, NILOP, Islamabad, Pakistan

Uteuliyev Yerzhan Sabitaliyevich

Department of Health Policy and Health Care Development, Kazakh Medical University of Continuing Education, Almaty, Kazakhstan

M. Sheeraz Ahmad

University Institute of Biochemistry and Biotechnology, Pir Mehr Ali Shah, Arid Agriculture University, Rawalpindi, Pakistan

Corresponding Author(s) : M. Sheeraz Ahmad

dr.sheeraz@uaar.edu.pk

Cellular and Molecular Biology, Vol. 66 No. 6: Issue 6
Article Published : September 30, 2020

Abstract

Surface functionalization of nanoparticles (NPs) for therapeutic siRNA delivery into cancer cells has gained interest. The present study was designed for surface functionalization of gold nanoparticles (AuNPs) for efficient siRNA delivery and knockdown in cancer cells. In order to achieve this objective, AuNPs were coated with HER2-siRNA in the presence of 11-mercaptoundecanoic acid (11-MUA), calcium chloride (CaCl2) and polyethyleneimine (PEI) in alternate charge bearing successive layers. MCF-7 cells were cultured and transfected with fabricated assembly of AuNPs. Cytotoxicity analysis revealed that the half inhibitory concentration (IC50) for the formulation was 45.35 nM . Total RNA was isolated from transfected cells, reverse transcribed into complementary DNA (cDNA) and real-time polymerase chain reaction (RT-PCR) was performed. The RT-PCR based delta-delta Ct analysis in treated cells revealed a significant 18.94 times decrease (p<0.001) in the expression of HER2 gene standardized with ACTB housekeeping gene as compared to untreated cells, which makes this formulation a potent approach for siRNA delivery and gene knockout.

Keywords

Gold nanoparticles HER2 siRNA silencing MCF-7.

Sami, A., Naqvi, S. S., Qayyum, M., Raza Rao, A., Sabitaliyevich, U. Y., & Ahmad, M. S. (2020). Calcium based siRNA coating: a novel approach for knockdown of HER2 gene in MCF-7 cells using gold nanoparticles. Cellular and Molecular Biology, 66(6), 105–111. https://doi.org/10.14715/cmb/2020.66.6.19