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Tocilizumab unmasks a stage-dependent interleukin-6 component in statin-induced apoptosis of metastatic melanoma cells.

Minichsdorfer C, Wasinger C, Sieczkowski E, Atil B, Hohenegger M - Melanoma Res. (2015)

Bottom Line: The effects of IL-6 were measured by western blots for STAT3 and Bcl-2 family proteins.This proapoptotic effect of IL-6 might be explained by a downregulation of Bcl-XL, observed only in WM35 cells.These results confirm that simvastatin facilitates apoptosis in combination with IL-6.

View Article: PubMed Central - PubMed

Affiliation: aDepartment of Oncology, Medicine I bCenter for Physiology and Pharmacology, Comprehensive Cancer Center, Institute of Pharmacology cDepartment of Neurology, Medical University of Vienna, Vienna, Austria.

No MeSH data available.


Related in: MedlinePlus

Tocilizumab unmasks an interleukin (IL)-6 contribution in simvastatin-induced caspase 3 activation. Melanoma cells (a) 518A2, (b) A375, and (c) WM35 were incubated in the absence [control (CTL)] and presence of 10 μmol/l simvastatin (Sim), human IgG1 (50 μg/ml), and/or tocilizumab (Toc; 50 μg/ml) for 48 h and the corresponding cell lysates used for caspase 3 assay (n=8–34). Statistical significance versus control: *P<0.05; **P<0.01; ***P<0.001; NS, not significant, or versus simvastatin treatment: #P<0.05; ##P<0.005.
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Figure 6: Tocilizumab unmasks an interleukin (IL)-6 contribution in simvastatin-induced caspase 3 activation. Melanoma cells (a) 518A2, (b) A375, and (c) WM35 were incubated in the absence [control (CTL)] and presence of 10 μmol/l simvastatin (Sim), human IgG1 (50 μg/ml), and/or tocilizumab (Toc; 50 μg/ml) for 48 h and the corresponding cell lysates used for caspase 3 assay (n=8–34). Statistical significance versus control: *P<0.05; **P<0.01; ***P<0.001; NS, not significant, or versus simvastatin treatment: #P<0.05; ##P<0.005.

Mentions: These findings are further supported by the observation that tocilizumab exerted no effect on caspase 3 activation in all three melanoma cell lines, which was also the case for the corresponding human IgG1 isoform (Fig. 6). Interestingly, a significant stimulation of caspase 3 was detected for the combination of simvastatin and IgG1 in WM35 cells versus control (Fig. 6c). Compared with simvastatin treatment alone, the combination of simvastatin plus IgG1 was insignificant. However, the simvastatin-induced caspase 3 activation was significantly reduced by tocilizumab in metastatic melanoma cells A375 and 518A2 (Fig. 6a and b). These cells were therefore used in scratch assays to elucidate a functional effect of tocilizumab-induced abrogation of simvastatin-induced apoptosis. Again, the human IgG1 conctrol exerted no effect on wound closure or simvastatin-induced inhibition (Fig. 7a and c). Obviously, proliferation in A375 and 518A2 cells was significantly prevented by simvastatin (Fig. 7). Although tocilizumab per se had no significant effect, coapplication with simvastatin resulted in accelerated reduction of the cell-free area, indicating accelerated proliferation. Hence, the latter observation confirmed abrogation of simvastatin-induced apoptosis by tocilizumab, which uncovered the involvement of proapoptotic IL-6 action in metastatic melanoma cells.


Tocilizumab unmasks a stage-dependent interleukin-6 component in statin-induced apoptosis of metastatic melanoma cells.

Minichsdorfer C, Wasinger C, Sieczkowski E, Atil B, Hohenegger M - Melanoma Res. (2015)

Tocilizumab unmasks an interleukin (IL)-6 contribution in simvastatin-induced caspase 3 activation. Melanoma cells (a) 518A2, (b) A375, and (c) WM35 were incubated in the absence [control (CTL)] and presence of 10 μmol/l simvastatin (Sim), human IgG1 (50 μg/ml), and/or tocilizumab (Toc; 50 μg/ml) for 48 h and the corresponding cell lysates used for caspase 3 assay (n=8–34). Statistical significance versus control: *P<0.05; **P<0.01; ***P<0.001; NS, not significant, or versus simvastatin treatment: #P<0.05; ##P<0.005.
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Figure 6: Tocilizumab unmasks an interleukin (IL)-6 contribution in simvastatin-induced caspase 3 activation. Melanoma cells (a) 518A2, (b) A375, and (c) WM35 were incubated in the absence [control (CTL)] and presence of 10 μmol/l simvastatin (Sim), human IgG1 (50 μg/ml), and/or tocilizumab (Toc; 50 μg/ml) for 48 h and the corresponding cell lysates used for caspase 3 assay (n=8–34). Statistical significance versus control: *P<0.05; **P<0.01; ***P<0.001; NS, not significant, or versus simvastatin treatment: #P<0.05; ##P<0.005.
Mentions: These findings are further supported by the observation that tocilizumab exerted no effect on caspase 3 activation in all three melanoma cell lines, which was also the case for the corresponding human IgG1 isoform (Fig. 6). Interestingly, a significant stimulation of caspase 3 was detected for the combination of simvastatin and IgG1 in WM35 cells versus control (Fig. 6c). Compared with simvastatin treatment alone, the combination of simvastatin plus IgG1 was insignificant. However, the simvastatin-induced caspase 3 activation was significantly reduced by tocilizumab in metastatic melanoma cells A375 and 518A2 (Fig. 6a and b). These cells were therefore used in scratch assays to elucidate a functional effect of tocilizumab-induced abrogation of simvastatin-induced apoptosis. Again, the human IgG1 conctrol exerted no effect on wound closure or simvastatin-induced inhibition (Fig. 7a and c). Obviously, proliferation in A375 and 518A2 cells was significantly prevented by simvastatin (Fig. 7). Although tocilizumab per se had no significant effect, coapplication with simvastatin resulted in accelerated reduction of the cell-free area, indicating accelerated proliferation. Hence, the latter observation confirmed abrogation of simvastatin-induced apoptosis by tocilizumab, which uncovered the involvement of proapoptotic IL-6 action in metastatic melanoma cells.

Bottom Line: The effects of IL-6 were measured by western blots for STAT3 and Bcl-2 family proteins.This proapoptotic effect of IL-6 might be explained by a downregulation of Bcl-XL, observed only in WM35 cells.These results confirm that simvastatin facilitates apoptosis in combination with IL-6.

View Article: PubMed Central - PubMed

Affiliation: aDepartment of Oncology, Medicine I bCenter for Physiology and Pharmacology, Comprehensive Cancer Center, Institute of Pharmacology cDepartment of Neurology, Medical University of Vienna, Vienna, Austria.

No MeSH data available.


Related in: MedlinePlus