Ulates DNA Repair Genes and Downregulates Genes associated to Hypoxia The involvement of PD-L1 with DNA repair, genomic instability, and hypoxia has not been extensively investigated. Here, we show that genes connected with DNA repair, MEN1 [29], and RNASEH1 [30], were upregulated following atezolizumab treatment (p 0.05. Figure 2E). In addition, atezolizumab upregulated the PAM16 gene, which could act as a tumor suppressor gene in addition to a regulator of ATP synthesis (p 0.01, Figure 2E). Notably, genes related to hypoxia/ATP synthesis or genes encoding heat shock proteins, including NDUFS3, ATP6V1G2, HSP90AB2P, HSPA8P3, HIGD1B, TK2, and ADCY1 (p 0.05, Figure 2E), and also genes related to the PI3K, MAPK, and NF-kB signaling pathways, which includes RASD1, MAP2K6, TRAF5, PIK3R2, C9orf96, and TENM1, had been drastically downregulated in treated cells (p 0.05, Figure 2F). In contrast, inhibitors for NF-kB activation, NFKBIB and COMMD6, were significantly up-regulated upon atezolizumab remedy (p 0.05, Figure 2F).Cancers 2019, 11, 1050 Cancers 2019, 11, x5 of5 ofFigure 2. Differentially expressed genes in MDA-MB-231 cells following atezolizumab therapy. Figure two. Differentially expressed genes in MDA-MB-231 cells following atezolizumab therapy. Hierarchical clustering of two independent experiments on differentially expressed RNA transcripts Hierarchical clustering of two independent experiments on differentially expressed RNA transcripts from RNA-Seq data. Each and every column represents a sample and every single row represents a transcript. Expression from RNA-Seq data. Each and every column represents a sample and every row represents a transcript. level of each and every gene in a single samplesingle sample is depicted as outlined by colour scale (A). show maps Expression level of every gene in a is depicted based on colour scale (A). Heat maps Heat the fold 1-Palmitoyl-2-oleoyl-sn-glycero-3-PC In Vivo modifications relative towards the mean expression of housekeeping genes (B), cell migration/metastasis/adhesion show the fold alterations relative for the mean expression of housekeeping genes (B), cell and EMT (C), anti-apoptosis, pro-apoptosis, and cell growth/proliferation pro-apoptosis, and hypoxia migration/metastasis/adhesion and EMT (C), anti-apoptosis, (D), DNA repair and cell (E), and signaling transduction (F). Final results hypoxia (E), and signaling transduction (F). Results are 1; growth/proliferation (D), DNA repair and are from two independent experiments. S1 = sample S2from two independent experiments. S1 = sample 1; S2 =with atezolizumab. = sample two; NT = non-treated cells; T = treated cells sample two; NT = non-treated cells; T = treated2.5. Atezolizumab Downregulates NF-kB, Akt, and CD40 Signaling Pathways2.5. Atezolizumab Downregulates NF-kB,the genes which had been downregulated in atezolizumab-treated Next, we located that about 19 of Akt, and CD40 Signaling Pathways cells are linked to EMT, 33 in the genes which had been downregulated in atezolizumab-treated Subsequent, we located that about 19 are associated to cell migration/invasion and metastasis, 16 are linked associated with transduction, favoring cell proliferation and EMT, 5 are anti-apoptotic, cells are with signaling EMT, 33 are related to cell migration/invasion and metastasis, 16 are eight are connected to signaling transduction, favoring cell proliferation 19 EMT, five are anti-apoptotic, linked to cell development and tumor cell proliferation, and and are related to hypoxia (Figure 3A). Selectedcell growth and tumor cell proliferation, and 19 panels, which includes CD40, MMP9, 8.