Fang Jinyong, Yi Kaining, Song Zaowen, Chen Mengyun, Xu Lingling, Zhao Tiejun
College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, Zhejiang, China.
Sheng Wu Gong Cheng Xue Bao. 2018 Sep 25;34(9):1491-1499. doi: 10.13345/j.cjb.170474.
We studied the effect of celastrol on the proliferation and apoptosis of adult T-cell leukemia (ATL) cells. After treating adult T-cell leukemia cell lines with different concentrations of celastrol, we analyzed the cell proliferation by MTT and colony formation assays. Flow cytometry was conducted to detect cell apoptosis by Annexin V-FITC/PI staining. Western blotting and dual-luciferase reporter assay were done to study the mechanism how celastrol suppressed the growth of adult T-cell leukemia cells. Celastrol could significantly inhibit the proliferation of adult T-cell leukemia cells, and induce apoptosis of ATL cells. With the increase of the concentration of celastrol, the ratio of Bax/Bcl-2 protein was up-regulated. The Caspase-3/7 protein was cleaved and activated after treatment with celastrol. Moreover, the expression of HTLV-1-encoded viral protein Tax was significantly inhibited in the celastrol treated cells. Taken together, these results indicated that celastrol effectively inhibited the proliferation of adult T-cell leukemia cells by regulating the expression of Bcl-2 family protein, and induced cell apoptosis by activating Caspase dependent pathway. In addition, celastrol could inhibit the expression of viral protein Tax. This study will provide an experimental basis for the clinical application of celastrol in the treatment of adult T-cell leukemia.
我们研究了雷公藤红素对成人T细胞白血病（ATL）细胞增殖和凋亡的影响。用不同浓度的雷公藤红素处理成人T细胞白血病细胞系后，我们通过MTT和集落形成试验分析细胞增殖情况。采用流式细胞术通过膜联蛋白V-异硫氰酸荧光素/碘化丙啶染色检测细胞凋亡。进行蛋白质免疫印迹法和双荧光素酶报告基因检测以研究雷公藤红素抑制成人T细胞白血病细胞生长的机制。雷公藤红素可显著抑制成人T细胞白血病细胞的增殖，并诱导ATL细胞凋亡。随着雷公藤红素浓度的增加，Bax/Bcl-2蛋白比值上调。用雷公藤红素处理后，半胱天冬酶-3/7蛋白被切割并激活。此外，在雷公藤红素处理的细胞中，人嗜T淋巴细胞病毒1型（HTLV-1）编码的病毒蛋白Tax的表达显著受到抑制。综上所述，这些结果表明雷公藤红素通过调节Bcl-2家族蛋白的表达有效抑制成人T细胞白血病细胞的增殖，并通过激活半胱天冬酶依赖性途径诱导细胞凋亡。此外，雷公藤红素可抑制病毒蛋白Tax的表达。本研究将为雷公藤红素在成人T细胞白血病治疗中的临床应用提供实验依据。
