Xu Cheng-Bo, Liao Bin, Shen Jian-Zhen, Fu Hai-Ying, Lin Ting
Department of Hematology, The People's Hospital Affiliated to Fujian University of Traditional Chinese Medicine, Fuzhou 350004.
Department of Hematology, The Union Hospital Affiliated to Fujian Medical University, Fujian Institute of Hematology, Fuzhou 350001, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2017 May 8;33(5):450-455. doi: 10.12047/j.cjap.5483.2017.108.
To investigate the effects of triptolide (Chinese Traditional Medicine)on the apoptosis and H3K4 protein methylation in multiple myeloma cells.
The RPMI8226 cells were cultured with different concentrations(10,20,40,80 and 160 nmol/L)of triptolide for different incubation time (24 h,48 h and 72 h). The inhibition of triptolide on RPMI8226 cell proliferation was detected by MTT assay. Apoptosis and cell cycle distribution were evaluated by flow cytometry.The expressions of H3K4me2 and trimethylation of histone H3 lysine 4(H3K4me3) in RPMI8226 cells were assayed by Western blot. The changes of expressions of histone methylase SET and MYND domain containing 3(SMYD3) and histone demethylase lysine specific demethylase 1(LSD1) in RPMI8226 cells were verified by qRT-PCR.
Triptolide had obvious inhibitive effects on proliferation of RPMI8226 cells and showed a dose-and time-dependent manner(<0.05). Triptolide induced apoptosis and G/M cell cycle arrest in a dose-dependent manner(<0.05). Triptolide decreased histone H3K4me2 and H3K4me3 expression in a dose-dependent manner(<0.05, <0.01). SMYD3 was significantly depressed at protein expression in a dose-dependent manner(<0.05), but LSD1 was up-regulated (<0.05).
Triptolide could inhibit RPMI8226 cell proliferation,induce the apoptosis and cause G/M arrest,meanwhile,significantly inhibit the protein expressions of H3K4me2 and H3K4me2 with alter the expression of SMYD3 and LSD1.The effects is probably related to the antitumor mechanism of MM cells induced by triptolide.
研究雷公藤甲素（一种中药）对多发性骨髓瘤细胞凋亡及H3K4蛋白甲基化的影响。
将RPMI8226细胞用不同浓度（10、20、40、80和160 nmol/L）的雷公藤甲素培养不同时间（24小时、48小时和72小时）。采用MTT法检测雷公藤甲素对RPMI8226细胞增殖的抑制作用。通过流式细胞术评估细胞凋亡和细胞周期分布。采用蛋白质印迹法检测RPMI8226细胞中H3K4me2和组蛋白H3赖氨酸4三甲基化（H3K4me3）的表达。通过qRT-PCR验证RPMI8226细胞中组蛋白甲基转移酶SET和含MYND结构域蛋白3（SMYD3）以及组蛋白去甲基化酶赖氨酸特异性去甲基化酶1（LSD1）表达的变化。
雷公藤甲素对RPMI8226细胞的增殖有明显抑制作用，呈剂量和时间依赖性（P<0.05）。雷公藤甲素以剂量依赖性方式诱导细胞凋亡和G/M期细胞周期阻滞（P<0.05）。雷公藤甲素以剂量依赖性方式降低组蛋白H3K4me2和H3K4me3的表达（P<0.05，P<0.01）。SMYD3蛋白表达呈剂量依赖性显著下调（P<0.05），但LSD1上调（P<0.05）。
雷公藤甲素可抑制RPMI8226细胞增殖，诱导细胞凋亡并导致G/M期阻滞，同时显著抑制H3K4me2和H3K4me3的蛋白表达，并改变SMYD3和LSD1的表达。其作用可能与雷公藤甲素诱导多发性骨髓瘤细胞的抗肿瘤机制有关。
