Increased tumorigenic activities of TIM-3–expressing RCC cells.

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Increased tumorigenic activities of TIM-3–expressing RCC cells. Increased tumorigenic activities of TIM-3–expressing RCC cells. A, ACHN cells transfected with human TIM-3 or control gene plasmid were treated with 50 nmol/L of sunitinib or rapamycin for 6 hours and then assayed for cell death by measuring active caspase-3. Similar results were observed in three experiments. B, bulk ACHN cells, TIM-3+ or TIM-3− ACHN cells, or those stimulated with TIM-3+ or TIM-3− CD14+ monocytes isolated from patients' PBMCs were then cultured with three passages in ultra-low attachment plates. The size (left) and numbers (right) of formed spheres generated per 100,000 cells were determined. C, mRNA expression of TIM-3, Twist1, Snail, and Pou5F in TIM-3+ or TIM-3− ACHN cells were quantified by RT-PCR. D, TIM-3+ or TIM-3− ACHN cells (104–105 per mouse) were inoculated s.c. into NSG mice, and tumor growth was measured. Similar results were obtained in three independent experiments. *, P < 0.05. ns, not statistically significant. Yoshihiro Komohara et al. Cancer Immunol Res 2015;3:999-1007 ©2015 by American Association for Cancer Research