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PTEN expression regulates autophagy in tumor cells, reducing T cell–mediated killing. PTEN expression regulates autophagy in tumor cells, reducing T cell–mediated.

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Presentation on theme: "PTEN expression regulates autophagy in tumor cells, reducing T cell–mediated killing. PTEN expression regulates autophagy in tumor cells, reducing T cell–mediated."— Presentation transcript:

1 PTEN expression regulates autophagy in tumor cells, reducing T cell–mediated killing.
PTEN expression regulates autophagy in tumor cells, reducing T cell–mediated killing. A, expression of LC3 I and LC3 II in BRAF-mutant melanoma cell lines with and without PTEN silencing. The expression levels of LC3 I and LC3 II in protein lysates from A375 and WM35 tumor cells were determined by Western blotting analysis. B, perturbing the expression of autophagy-related genes changes the sensitivity of melanoma to apoptosis induced by tumor-reactive T cells. A patient-derived melanoma cell line, Mel2400, was transduced with lentiviral vectors encoding the open reading frames (ORF) or shRNAs of autophagy-related genes. Virally transduced tumor cells were cocultured with paired autologous T cells for 3 hours. The percentage of killed (cleaved casp-3+) tumor cells expressing ORF or shRNAs was determined using flow cytometry. A comboscore was calculated as described in the Methods section and used to evaluate the effect of genetic modifications on sensitivity of tumor cells to T cell–mediated killing. Tumor cells transduced with virus expressing GFP or shRNA targeting luciferase (Luc) served as controls for overexpression and knockdown experiments, respectively. *, genetic modification, which significantly changed the sensitivity of tumor to T cell–mediated killing (P < 0.05). C, PTEN-silenced and control melanoma cells were transduced with a viral vector encoding MAP1LC3B. The percentage of cleaved casp-3+ cells among transduced tumor cells in response to PMEL-1 T cells (E:T = 10:1) was evaluated. *, P < 0.05; **, P < 0.01; ****, P < D, melanoma cells (Mel2338) were pretreated with 1 μmol/L hydroxychloroquine (HCQ) overnight and followed by coculture with paired TILs for 3 hours. The percentage of apoptotic tumor cells was evaluated by the cleaved caspase-3 assay. Results are representative of data generated in two independent experiments. Weiyi Peng et al. Cancer Discov 2016;6: ©2016 by American Association for Cancer Research

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