Figure 5 from Selective Depletion of Cancer Cells with Extrachromosomal DNA via Lentiviral Infection

Low transduction efficiency and lethal effect of lentivirus in ecDNA+ cells lead to selective depletion. A, Differential lentiviral transduction efficiency in PC3 single-cell clones with MYC-ecDNA (EC) or MYC-HSR (HSR). Two single-cell clones per MYC amplification category were tested. Cells were transduced with lentivirus carrying green fluorescence protein (GFP, genome size = 6.4 kb) for 24 hours and subjected to flow cytometry analysis. Lentiviral transduction efficiency was calculated by quantifying the proportion of cells expressing GFP (t test, *, P < 0.05; ns = 3). B, Cell proliferation comparison between PC3 single-cell clones with EC or HSR. Two single-cell clones per MYC amplification category were tested. Five sets of cells were transduced with lentivirus carrying red fluorescence protein (mCherry, genome size = 4.5 kb), and each set of cells was subjected to the CellTiter-Glo assay on different days (1, 2, 3, 4, 5 days after transduction). The posttransduction proliferation rate was compared with the proliferation rate of the untreated control cells. C, Cell death analysis was performed in two isogenic pairs of PC3 clones. Cell death of each clone after 48 hours of lentivirus transduction (genome size = 4.5 kb) was analyzed by propidium iodide (PI) staining and compared with the nontransduced control cells (t test, *, P < 0.001; nd = 3 per clone).