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Am J Physiol Cell Physiol 281: C1812-C1818, 2001;
0363-6143/01 $5.00
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Vol. 281, Issue 6, C1812-C1818, December 2001

B16-BL6 melanoma cells release inhibitory factor(s) of active pump activity in isolated lymph vessels

Kei Nakaya1, Risuke Mizuno1, and Toshio Ohhashi1,2

1 First Department of Physiology, Shinshu University School of Medicine, and 2 Institute of Organ Transplants, Reconstructive Medicine, and Tissue Engineering, Shinshu University Graduate School of Medicine, Matsumoto 390-8621, Japan

We investigated whether supernatant cultured with melanoma cell lines B16-BL6 and K1735 or the Lewis lung carcinoma cell line (LLC) can regulate lymphatic pump activity with bioassay preparations isolated from murine iliac lymph vessels. B16-BL6 and LLC supernatants caused significant dilation of lymph microvessels with cessation of pump activity. B16-BL6 supernatant produced dose-related cessation of lymphatic pump activity. There was no significant tachyphylaxis in the supernatant-mediated inhibitory response of lymphatic pump activity. Pretreatment with 3 × 10-5 M Nomega -nitro-L-arginine methyl ester (L-NAME) or 10-7 M or 10-6 M glibenclamide and 5 × 10-4 M 5-hydroxydecanoic acid caused significant reduction of supernatant-mediated inhibitory responses. Simultaneous treatment with 10-3 M L-arginine and 3 × 10-5 M L-NAME significantly lessened L-NAME-induced inhibition of the supernatant-mediated response, suggesting that endogenous nitric oxide (NO) plays important roles in supernatant-mediated inhibitory responses. Chemical treatment dialyzed substances of <1,000 molecular weight (MW), producing complete reduction of the supernatant-mediated response. In contrast, pretreatment with heating or digestion with protease had no significant effect on supernatant-mediated response. These findings suggest that B16-BL6 cells may release nonpeptide substance(s) of <1,000 MW, resulting in significant cessation of lymphatic pump activity via production and release of endogenous NO and activation of mitochondrial ATP-sensitive K+ channels.

malignant melanoma; active lymph transport; nitric oxide; mitochondrial adenosine 5'-triphosphate-sensitive potassium channel


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