Abstract

Alteronol, isolated from microbial mutation strains, has been applied for Chinese and International patents for tumor treatment. The aim of this project study is to investigate characteristics of proliferation and redifferentiation induced by alteronol in B16-F0 mouse melanoma cells. Cell proliferation is determined by tetrazolium salt colorimetric method (MTT assay). Morphological changes were analyzed by using Giemsa staining. The levels of melanin and tyrosinase were measured by spectrophotometry. The mRNA expressions of tyrosinase-related protein Trp1 and Trp2 were evaluated by reverse transcription-polymerase chain reaction (RT-PCR). The anchorage-independent proliferation of B16-F0 was monitored by the colony formation assay. Tumorigenicity was characterized by an animal model in vivo. The results showed that the proliferation of B16-F0 cells was inhibited by alteronol in a concentration and time dependent manner. All well-known evaluation indexes of melanoma cell differentiation, including morphological changes and tyrosinase activity alteration, were greatly enhanced with the increase of alteronol concentrations. Taken together, the expression of tyrosinase related gene, decreased cell colony formation rate and the tumorigenicity in vivo; all of these revealed that alteronol plays a key role in inducing differentiation and suppressing the proliferation of B16-F0 tumor cells in vitro and in vivo.

Keywords: Alteronol, B16F0, differentiation, melanin, tyrosinase, tumorigenicity.