AbstractPurpose
The goal of this study is to explore the effects of columbamine in melanoma cells and the signaling pathway involved.
Methods
Human melanoma cell line A375 cells were used in this study. Cell proliferative ability was detected by MTT assay and clone formation assay. Cell migration and invasion were measured by wound healing assay and transwell assay, respectively. Protein expression was examined by Western blotting.
Results
Columbamine reduced cell proliferative ability and the number of clone spots in A375 cells. Western blotting results demonstrated that expression of cleaved caspase 3, an activated cell death protease, was upregulated by 20 and 50 µM of columbamine. Wound healing results showed that the scratch width was wider in cell treated with 20 and 50 µM of columbamine than that in cell treated with 0 and 10 µM of columbamine. Phosphorylation of STAT3 and expression of HSP90 was also repressed by columbamine in a concentration-dependent manner. Overexpression of HSP90 attenuated the inhibition of cell proliferation, migration and invasion induced by columbamine.
Conclusion
Columbamine inhibited melanoma cell proliferation, migration, and invasion in A375 cells through inactivation of STAT3, which is mediated by HSP90.
… Louis, MO) and 1% penicillin–streptomycin (100 μg/mL; Invitrogen, Carlsbad, CA)
at 37 °C with 95% air and 5% CO 2 . Columbamine (Biopurify, China) was
dissolved in ethanol (Worldyachem, Shanghai, China). A375 …
