China Medical University, Shenyang, 110001, China; Department of Dermatology, No.1 Hospital of China Medical University, China; Key Laboratory of Immunodermatology, Ministry of Health and Ministry of Education, Shenyang, 110001, China.
School of Medicine, Tsinghua University, Beijing, 100084, China; China Medical University, Shenyang, 110001, China.
J Dermatol Sci. 2018 Sep;91(3):256-267. doi: 10.1016/j.jdermsci.2018.05.006. Epub 2018 May 23.
Hyperthermia is an effective treatment against cancer and human papillomavirus (HPV) infection. Previous studies have shown that heat shock proteins are crucial to the action of hyperthermia.
To examine the effects of hyperthermia in combination with DNAJA4-deficiency on human keratinocytes and Condyloma acumunatum (CA) tissues.
HaCaT cells were subjected to 44°C (compared to 37°C) waterbath for 30min for stimulation. Foreskin or CA tissues obtained from patients undergoing circumcision or pathological examination were bisected and subjected to similar treatments. DNAJA4-knockout (KO) HaCaT cells were generated with CRISPR/Cas9 technology. mRNA and protein expressions were determined using rt-qPCR and western-blotting. Cell cycle distribution, apoptosis and senescence were analyzed by flow cytometry.
DNAJA4 was induced in HaCaT cells, foreskin and CA tissues subjected to hyperthermia at both transcriptional and translational levels. NF-kB, was activated by hyperthermia in HaCaT cells, and further enhanced by DNAJA4-deficiency. Transcription of TNF-α; IL-1B, TNFAIP3 and IL-8 were induced in HaCaT cells subjected to hyperthermia. DNAJA4-knockout promoted transcriptions of TNF-α and IL-1B, whereas decreased that of TNFAIP3 and IL-8. Reduced cell survival, proliferation and viability were demonstrated using flow cytometry and MTS assays. Furthermore, NF-kB inhibitors reversed most of the phenotypes observed.
Hyperthermia reduced HaCaT cell proliferation and promoted cytokine expressions responsible for anti-viral activity, mainly through a NF-kB dependent pathway. DNAJA4-deficiency enhanced the activation of NF-kB by hyperthermia in HaCaT cells, indicating that DNAJA4 may be a promising therapeutic target for use in the treatment of cutaneous HPV infections.
热疗是一种有效的癌症和人乳头瘤病毒（HPV）感染治疗方法。先前的研究表明热休克蛋白对于热疗的作用至关重要。
研究热疗联合 DNAJA4 缺乏对人角质形成细胞和尖锐湿疣（CA）组织的影响。
HaCaT 细胞在 44°C（与 37°C 相比）水浴中刺激 30min。从接受包皮环切术或病理检查的患者中获得包皮或 CA 组织，并进行类似处理。利用 CRISPR/Cas9 技术生成 DNAJA4 敲除（KO）HaCaT 细胞。采用 rt-qPCR 和 Western-blotting 检测 mRNA 和蛋白表达。通过流式细胞术分析细胞周期分布、凋亡和衰老。
热疗可诱导 HaCaT 细胞、包皮和 CA 组织中 DNAJA4 的转录和翻译。NF-kB 在 HaCaT 细胞中被热疗激活，且 DNAJA4 缺乏进一步增强了 NF-kB 的激活。TNF-α；IL-1B、TNFAIP3 和 IL-8 在热疗处理的 HaCaT 细胞中转录。DNAJA4 敲除促进了 TNF-α和 IL-1B 的转录，而降低了 TNFAIP3 和 IL-8 的转录。利用流式细胞术和 MTS 分析，证明了 DNAJA4 敲除降低了细胞的存活、增殖和活力。此外，NF-kB 抑制剂逆转了大部分观察到的表型。
热疗降低了 HaCaT 细胞的增殖，并促进了抗病毒活性相关细胞因子的表达，主要通过 NF-kB 依赖途径。DNAJA4 缺乏增强了 HaCaT 细胞中热疗对 NF-kB 的激活，表明 DNAJA4 可能是治疗皮肤 HPV 感染的有前途的治疗靶点。
