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Quantitative proteomic analysis uncovers inhibition of melanin synthesis by silk fibroin via MITF/tyrosinase axis in B16 melanoma cells
Life Sciences ( IF 5.2 ) Pub Date : 2021-09-01 , DOI: 10.1016/j.lfs.2021.119930 Yuqiu Wang 1 , Tianbi Duan 2 , Minhua Hong 2 , Yanting Zhou 3 , Hui Huang 3 , Xiao Xiao 4 , Jing Zheng 5 , Hu Zhou 6 , Zhi Lu 2
Life Sciences ( IF 5.2 ) Pub Date : 2021-09-01 , DOI: 10.1016/j.lfs.2021.119930 Yuqiu Wang 1 , Tianbi Duan 2 , Minhua Hong 2 , Yanting Zhou 3 , Hui Huang 3 , Xiao Xiao 4 , Jing Zheng 5 , Hu Zhou 6 , Zhi Lu 2
Affiliation
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Silk fibroin (SF), a natural product from silkworms, has been used to promote anti-inflammation, induce wound healing, and reduce melanin production. However, the underlying regulatory mechanism of SF on melanin production remains unknown. The aim of this study was to investigate the distinct regulatory mechanism of SF in B16 melanoma cells by applying quantitative proteomic approach. B16 melanoma cells were treated with PBS, KA or SF for 48 h, respectively. Cell viability, melanin content, and tyrosinase activity were examined. A label-free quantitative proteomic approach was utilized to investigate the regulatory mechanism of SF. The differentially expressed proteins and their related biological processes were subsequently identified by bioinformatics methods. Furthermore, the identified differentially expressed proteins were validated by western blot. Both SF and KA were able to suppress the melanin synthesis of B16 melanoma cells without appreciable toxicity; yet, SF had a distinct effect on mushroom tyrosinase activity in vitro. Moreover, quantitative proteomic approach identified 141 proteins differentially expressed only in SF/Con group. Bioinformatic analysis of these proteins revealed that oxidation-reduction process, RNA processing, fatty acid degradation, as well as melanin biosynthetic process were enriched with SF treatment. The proteins associated with melanin biosynthetic process, including microphthalmia-associated transcription factor (MITF) and tyrosinase, were down-regulated in SF group, which was confirmed by western blot. SF inhibited melanin synthesis in B16 melanoma cells via down-regulation of MITF and tyrosinase expression, which provides a rationale for future utilization of SF.
中文翻译：
定量蛋白质组学分析揭示丝素蛋白通过 MITF/酪氨酸酶轴抑制 B16 黑色素瘤细胞中的黑色素合成
丝素蛋白（SF）是蚕的天然产物，已被用于促进抗炎、诱导伤口愈合和减少黑色素的产生。然而，SF对黑色素产生的潜在调节机制仍不清楚。本研究的目的是通过应用定量蛋白质组学方法研究 SF 在 B16 黑色素瘤细胞中的独特调节机制。 B16 黑色素瘤细胞分别用 PBS、KA 或 SF 处理 48 小时。检查细胞活力、黑色素含量和酪氨酸酶活性。采用无标记定量蛋白质组学方法研究 SF 的调控机制。随后通过生物信息学方法鉴定差异表达的蛋白质及其相关的生物过程。此外，通过蛋白质印迹验证了鉴定的差异表达蛋白。 SF和KA均能够抑制B16黑色素瘤细胞的黑色素合成，且没有明显的毒性；然而，SF对体外蘑菇酪氨酸酶活性有明显影响。此外，定量蛋白质组学方法鉴定出仅在 SF/Con 组中差异表达的 141 个蛋白质。对这些蛋白质的生物信息分析表明，SF 处理丰富了氧化还原过程、RNA 加工、脂肪酸降解以及黑色素生物合成过程。与黑色素生物合成过程相关的蛋白质，包括小眼相关转录因子（MITF）和酪氨酸酶，在SF组中表达下调，蛋白质印迹证实了这一点。 SF 通过下调 MITF 和酪氨酸酶表达来抑制 B16 黑色素瘤细胞中黑色素的合成，这为 SF 的未来利用提供了理论依据。
更新日期：2021-09-01
中文翻译：
定量蛋白质组学分析揭示丝素蛋白通过 MITF/酪氨酸酶轴抑制 B16 黑色素瘤细胞中的黑色素合成
丝素蛋白（SF）是蚕的天然产物，已被用于促进抗炎、诱导伤口愈合和减少黑色素的产生。然而，SF对黑色素产生的潜在调节机制仍不清楚。本研究的目的是通过应用定量蛋白质组学方法研究 SF 在 B16 黑色素瘤细胞中的独特调节机制。 B16 黑色素瘤细胞分别用 PBS、KA 或 SF 处理 48 小时。检查细胞活力、黑色素含量和酪氨酸酶活性。采用无标记定量蛋白质组学方法研究 SF 的调控机制。随后通过生物信息学方法鉴定差异表达的蛋白质及其相关的生物过程。此外，通过蛋白质印迹验证了鉴定的差异表达蛋白。 SF和KA均能够抑制B16黑色素瘤细胞的黑色素合成，且没有明显的毒性；然而，SF对体外蘑菇酪氨酸酶活性有明显影响。此外，定量蛋白质组学方法鉴定出仅在 SF/Con 组中差异表达的 141 个蛋白质。对这些蛋白质的生物信息分析表明，SF 处理丰富了氧化还原过程、RNA 加工、脂肪酸降解以及黑色素生物合成过程。与黑色素生物合成过程相关的蛋白质，包括小眼相关转录因子（MITF）和酪氨酸酶，在SF组中表达下调，蛋白质印迹证实了这一点。 SF 通过下调 MITF 和酪氨酸酶表达来抑制 B16 黑色素瘤细胞中黑色素的合成，这为 SF 的未来利用提供了理论依据。
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