Department of Dermatology, Zhejiang Hospital of Traditional Chinese Medicine, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China (mainland).
Department of Dermatology, The Third People's Hospital in Hangzhou Affiliated to Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China (mainland).
Med Sci Monit. 2019 Jan 15;25:436-442. doi: 10.12659/MSM.913889.
BACKGROUND Radix Tetrastigma Hemsleyani Flavone (RTHF) has detoxification and anti- inflammation activity and is widely used. Here, we report that RTHF inhibits cell proliferation and induces apoptosis in cutaneous squamous cell carcinoma A431 cells and is a potential strategy for cancer therapy. MATERIAL AND METHODS A431 cells were cultured in different concentrations of RTHF. The inhibition of cell proliferation was assessed by MTT assay, cell apoptosis was shown through FCM, and cell invasion was assessed by Transwell methods. Enzyme proteasome assay was used to detect the activity of proteasome and DUB. Expression of apoptosis-related and ubiquitin proteasome pathway-associated proteins were assessed by PCR and Western blot. RESULTS RTHF obviously suppressed the proliferation and induced apoptosis of A431 cells in a dose-dependent manner. Transwell assay showed that RTHF inhibited the cell metastasis significantly. Enzyme proteasome assay show that the RTHF treatment of activity of proteasome and DUB was significantly lower than in control. RTHF increased the expression of Bax and inhibited Bcl-2, pro-caspase3, and pro-caspase9 activity. The expression of USP14, UCHL5, and POH1 decreased and ub-prs increased significantly in the treatment group. CONCLUSIONS Our study reveals that RTHF-mediated inhibition of DUBs and proteasome may provide a potential strategy for cancer therapy.
三叶木通黄酮(RTHF)具有解毒和抗炎活性,应用广泛。本文报道 RTHF 可抑制皮肤鳞状细胞癌细胞 A431 增殖并诱导其凋亡,是癌症治疗的一种潜在策略。
将 A431 细胞在不同浓度的 RTHF 中培养。通过 MTT 检测抑制细胞增殖,FCM 显示细胞凋亡,Transwell 法检测细胞侵袭。用酶蛋白酶体测定法检测蛋白酶体和 DUB 的活性。通过 PCR 和 Western blot 评估凋亡相关和泛素蛋白酶体途径相关蛋白的表达。
RTHF 呈剂量依赖性明显抑制 A431 细胞增殖并诱导其凋亡。Transwell 检测显示 RTHF 显著抑制细胞转移。酶蛋白酶体测定显示,RTHF 处理组的蛋白酶体和 DUB 活性明显低于对照组。RTHF 增加 Bax 的表达并抑制 Bcl-2、原胱天蛋白酶 3 和原胱天蛋白酶 9 的活性。处理组中 USP14、UCHL5 和 POH1 的表达显著降低,ub-prs 明显增加。
本研究揭示了 RTHF 介导的 DUBs 和蛋白酶体抑制可能为癌症治疗提供一种潜在策略。