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. 2021 Dec 8;9(12):e002772. doi: 10.1136/jitc-2021-002772

Figure 3.

Figure 3

SN-38 or metformin significantly suppresses tumor growth in vivo monitored by bioluminescent imaging. (A) Timeline (weekly) of bioluminescent signals in a representative showed growing mouse tumor injected with 5.0×106 ID8-luc2 cells. (B) In the DMSO control group, he bioluminescence level increased significantly at weeks 4 and 5 (*p<0.05, **p<0.005). All scales in photons/second/cm2/steradian. (C) This figure shows flow cytometry analysis of tumor cells treated with controls or two different drugs (DMSO, metformin, or SN-38). Gray color: isotype control; blue color: DMSO-treated tumor cells; green color: metformin-treated tumor cells; and orange color: SN-38-treated tumor cells. The right panel shows elevated FOXO3 expression on each drug-treated tumor cells (n=3, *p<0.05, **p<0.005). (D) Gray color: isotype control; blue color: DMSO-treated tumor cells; green color: metformin-treated tumor cells; and orange color: SN-38-treated tumor cells. The right panel shows decreased PD-L1 expression on each drug-treated tumor cells (n=3, *p<0.05, **p<0.005). (E) This figure shows flow cytometry analysis of CD3 or NKp46 expressing immune cells isolated from mouse tumors. CD3 (T cell marker) or NKp46 (NK cell marker) shows each changed population in the drug-treated tumor. Metformin or SN-38 treated tumor tissues show increased CD3 positive cells or NKp46 positive cells. The right panel shows a significantly increased CD3 or NKp46 positive population in tumor tissues (n=3, *p<0.05, **p<0.005). (F) This figure shows flow cytometry analysis of CD8 or NKp46 expressing immune cells isolated from mouse tumors. CD8 (cytotoxic T cell marker) or NKp46 (NK cell marker) shows each changed population in the drug-treated tumor. Metformin or SN-38 treated tumor tissues show increased CD8 positive cells or NKp46 positive cells. The right panel shows a significantly increased CD8 (cytotoxic T cells) positive population in tumor tissues (n=3, *p<0.05). DMSO, dimethylsulfoxide; NK, natural killer; PD-L1, programmed death-ligand 1.