Effects of RNA interference-mediated NRP-1 silencing on the proliferation and apoptosis of breast cancer cells

Molecular Medicine Reports
Zhengxiang HanXiuping Du

Abstract

Lentiviral expression vectors carrying human NRP-1 short hairpin RNA (shRNA) were constructed and selected to present highly efficient NRP-1/shRNA interference sequences, in order to investigate the effects of RNA interference (RNAi)-mediated NRP-1 silencing on the biological activities of breast cancer cells. Three pairs of human NRP-1 targeted specific interference sequences and one pair of non-specific control sequences were designed, synthesized and subcloned into pLB lentiviral vectors, which were further identified by polymerase chain reaction (PCR) and sequencing. Recombinant and lentiviral packaging plasmids were co-transfected into 293FT cell lines in order to produce lentiviral particles and to infect breast cancer cells with high NRP-1 expression. Flow cytometry was used to sort green fluorescent protein-positive cells. Fluorescence quantitative-reverse transcription-PCR and western blot analysis were employed to identify the interference silencing sequence with the most efficient silencing profile. A cell counting kit-8 assay and an Annexin V-propidium iodide method in combination with flow cytometry were used to examine the effects of RNA interference-mediated NRP-1 gene silencing on cell proliferation, apoptosis a...Continue Reading

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Citations

Jul 6, 2018·Endocrine-related Cancer·Andrea NicoliniAngelo Carpi
Jun 18, 2016·International Journal of Oncology·Xiaofeng DouXinhui Su
Jun 13, 2017·Expert Opinion on Biological Therapy·Antonela S AsadMarianela Candolfi
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