Human epidermal melanocytes can be induced to form melanocyte spheroids and revert to immature characteristics by long-term trypsinization (LTT). To further explore the biological characteristics of melanocytes after LTT and to study the underlying mechanism. Melanocytes were subjected to long-term (2 h) trypsinization in this study, after which their viability, proliferation and autophagy were characterized. The expression of melanocyte markers [human melanoma black45 (HMB45), tyrosinase (TYR) and Nestin] was detected and relative expression levels of mRNAs encoding TYR, Nestin, c-Kit and microphthalmia-associated transcription factor (MITF) were determined. After LTT, more short spindle-shaped melanocytes appeared and viability assays demonstrated that most melanocytes survived that treatment but had decreased proliferation rates compared to the untreated controls. There was a significant increase in autophagy of melanocytes after LTT and the expression of TYR was decreased compared with untreated control melanocytes. There were no significant differences in the expression of HMB45 or Nestin between the two groups. Compared with untreated melanocytes, levels of message ribonucleic acid (mRNAs) encoding TYR, c-Kit and MITF were decreased after LTT, while Nestin mRNA levels were increased. These results clarified that Long-term treatment with trypsin causes the dedifferentiation of mature epidermal melanocytes in vitro.

人表皮黑色素细胞可以被诱导形成黑色素细胞球体，并通过长期胰蛋白酶消化 (LTT) 恢复到不成熟的特征。进一步探索LTT后黑色素细胞的生物学特性并研究其潜在机制。在本研究中，对黑色素细胞进行长期（2 小时）胰蛋白酶消化，然后对它们的活力、增殖和自噬进行表征。检测黑色素细胞标志物[人黑色素瘤 black45 (HMB45)、酪氨酸酶 (TYR) 和 Nestin] 的表达，并测定编码 TYR、Nestin、c-Kit 和小眼相关转录因子 (MITF) 的 mRNA 的相对表达水平。LTT之后，出现了更多的短纺锤形黑素细胞，活力测定表明，大多数黑素细胞在该处理中存活下来，但与未处理的对照相比，增殖率降低。与未处理的对照黑素细胞相比，LTT 后黑素细胞的自噬显着增加，TYR 的表达降低。两组间HMB45或Nestin的表达无显着差异。与未处理的黑素细胞相比，LTT 后编码 TYR、c-Kit 和 MITF 的信息核糖核酸 (mRNA) 水平降低，而 Nestin mRNA 水平增加。这些结果阐明了长期用胰蛋白酶处理会导致体外成熟表皮黑素细胞的去分化。与未处理的对照黑素细胞相比，LTT 后黑素细胞的自噬显着增加，TYR 的表达降低。两组间HMB45或Nestin的表达无显着差异。与未处理的黑素细胞相比，LTT 后编码 TYR、c-Kit 和 MITF 的信息核糖核酸 (mRNA) 水平降低，而 Nestin mRNA 水平增加。这些结果阐明了长期用胰蛋白酶处理会导致体外成熟表皮黑素细胞的去分化。与未处理的对照黑素细胞相比，LTT 后黑素细胞的自噬显着增加，TYR 的表达降低。两组间HMB45或Nestin的表达无显着差异。与未处理的黑素细胞相比，LTT 后编码 TYR、c-Kit 和 MITF 的信息核糖核酸 (mRNA) 水平降低，而 Nestin mRNA 水平增加。这些结果阐明了长期用胰蛋白酶处理会导致体外成熟表皮黑素细胞的去分化。LTT 后 c-Kit 和 MITF 减少，而 Nestin mRNA 水平增加。这些结果阐明了长期用胰蛋白酶处理会导致体外成熟表皮黑素细胞的去分化。LTT 后 c-Kit 和 MITF 减少，而 Nestin mRNA 水平增加。这些结果阐明了长期用胰蛋白酶处理会导致体外成熟表皮黑素细胞的去分化。