Interference of Hsp27 Results in Apoptosis Induced by Photodamage via Regulation of Subcellular Localization of p21 in Immortalized Human Keratinocytes
Interference of Hsp27 Results in Apoptosis Induced by Photodamage via Regulation of Subcellular Localization of p21 in Immortalized Human Keratinocytes
复制标题
通过调节永生化人角质形成细胞中 p21 的亚细胞定位来干扰 Hsp27 导致光损伤诱导的细胞凋亡
DOI：
10.12659/msm.917164
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发表时间：
2019-10-08
影响因子：
3.1
通讯作者：
Chen, Ai-Jun
中科院分区：
文献类型：
--
作者：
Liu, Chuan;Huang, Xin;Chen, Ai-Jun
背景：近年来，由于紫外光引起的光性皮肤病发病率增加，有必要阐明潜在的皮肤光损伤机制并揭示可能的治疗靶点。热休克蛋白27 （Hsp27）具有抑制细胞凋亡的作用。本研究的目的是阐明Hsp27和p21对紫外线B （UVB）诱导的光损伤可能的光保护机制。材料/方法：对Hsp27基因进行干扰，评估其下游效应物的表达、细胞凋亡和细胞增殖能力。流式细胞术检测细胞凋亡。采用细胞计数试剂盒-8 （CCK-8）法检测细胞增殖能力。western-blotting法检测蛋白表达。采用定量反转录聚合酶链反应（qRT-PCR）检测mRNA的表达。亚细胞定位用免疫荧光法进行鉴定。结果：Hsp27敲低可降低细胞活力，增加uvb诱导的细胞凋亡发生率。与对照组相比，Hsp27敲低组磷酸化akt （p-Akt）依赖通路的激活导致p21在细胞核内聚集，抑制细胞增殖，同时促进细胞凋亡。此外，Hsp27敲低使p53表达升高，Bax: Bcl-2比值升高，进一步加速了细胞凋亡过程。结论：这些发现补充了皮肤光损伤的机制，证明了Hsp27在HaCaT细胞中的光保护作用，可能提示了光损伤和光性皮肤病的潜在治疗靶点。
Background: Owing to the increased incidence of photodermatosis caused by ultraviolet light in recent years, it is necessary to clarify the mechanisms potential photodamage to the skin and reveal possible therapeutic targets. Heat shock protein 27 (Hsp27) is well known for suppressing apoptosis. The aim of present study was to elucidate possible photoprotective mechanism between Hsp27 and p21 on ultraviolet B (UVB)-induced photodamage.Material/Methods: The Hsp27 gene was interfered to assess the expression of its downstream effectors, cell apoptosis, and cell proliferation ability. The cell apoptosis was tested using flow cytometry method. The cell proliferation ability was tested using Cell Counting Kit-8 (CCK-8) assay. The expression of protein was tested using western-blotting method. The expression of mRNA was detected using quantitative reverse transcription polymerase chain reaction (qRT-PCR). The subcellular localization was elucidated using immunofluorescence.Results: Hsp27 knockdown decreased cell viability and increased the incidence of UVB-induced apoptosis. Compared with control group, activation of phosphorylated-Akt (p-Akt)-dependent pathway resulted in the nuclear accumulation of p21 and suppression of cell proliferation, while promoting apoptosis in Hsp27 knockdown group. In addition, Hsp27 knockdown increased p53 expression and the Bax: Bcl-2 ratio, which further accelerated the apoptotic process.Conclusions: These findings complemented the mechanism of skin photodamage and demonstrated the photoprotective mechanisms of Hsp27 in HaCaT cells, which might implicate a potential therapeutic target of photodamage and photodermatosis.