Department of Burn and Plastic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
J Cosmet Dermatol. 2020 May;19(5):1224-1230. doi: 10.1111/jocd.13126. Epub 2019 Sep 4.
To investigate the relationship among natural aging of epidermal cells, epigenetics, and SPRY1 methylation mechanism.
Immunohistochemistry, reverse transcription-PCR (RT-PCR), and Western blot were used to detect the expression of DNA methyltransferase 1 (DNMT1) and Sprouty1 (SPRY1) in skin epithelial cells from different age groups. An aging model of HaCaT cells was constructed. In HaCaT cells and their aging groups, DNMT1 and SPRY1 expression were detected by RT-PCR and WB. SPRY1 methylation status in epidermal cells from different age groups and HaCaT cells were detected by Methylation-Specific PCR (MS-PCR).
The expression of DNMT1 and SPRY1 in skin epithelial cells from natural aging groups decreased with age; there was no significant difference in the expression of DNMT1 in HaCaT cells and the different age groups. The expression of SPRY1 in HaCaT cells was lower than it was in the aging groups. The methylation status of SPRY1 gradually decreased as the age of skin epidermal cells increased, while the methylation status of SPRY1 was not different between HaCaT cells and the aging group.
DNMT1 is involved in the regulation of natural aging of skin epidermal cells but has a nominal role in our induced aging model. SPRY1 is involved in natural aging and induced aging of skin epidermal cells. The regulation of SPRY1 methylation is involved in the natural senescence of skin epidermal cells, while the induced aging of epidermal cells is nominally involved in the mechanism of SPRY1 methylation.
研究表皮细胞自然衰老、表观遗传学与 SPRY1 甲基化机制之间的关系。
采用免疫组化、逆转录-PCR（RT-PCR）和 Western blot 检测不同年龄组皮肤上皮细胞中 DNA 甲基转移酶 1（DNMT1）和 Sprouty1（SPRY1）的表达。构建 HaCaT 细胞衰老模型。在 HaCaT 细胞及其衰老组中，通过 RT-PCR 和 WB 检测 DNMT1 和 SPRY1 的表达。采用甲基化特异性 PCR（MS-PCR）检测不同年龄组表皮细胞和 HaCaT 细胞中 SPRY1 的甲基化状态。
自然衰老组皮肤上皮细胞中 DNMT1 和 SPRY1 的表达随年龄增长而降低；HaCaT 细胞与不同年龄组中 DNMT1 的表达无明显差异。HaCaT 细胞中 SPRY1 的表达低于衰老组。随着皮肤表皮细胞年龄的增加，SPRY1 的甲基化状态逐渐降低，而 HaCaT 细胞与衰老组之间的 SPRY1 甲基化状态无差异。
DNMT1 参与皮肤表皮细胞的自然衰老调节，但在我们诱导的衰老模型中作用不大。SPRY1 参与皮肤表皮细胞的自然衰老和诱导衰老。SPRY1 甲基化的调节参与皮肤表皮细胞的自然衰老，而表皮细胞的诱导衰老在 SPRY1 甲基化机制中作用不大。
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