冬凌草甲素通过下调Brg1表达抑制Jurkat细胞生长

叶珍珍; 薛飞龙; 丁文评; 孔祥; 沈伊娜

doi:10.7499/j.issn.1008-8830.2017.11.016

PDF(1312 KB)

HTML

中国当代儿科杂志 ›› 2017, Vol. 19 ›› Issue (11) : 1208-1212. DOI: 10.7499/j.issn.1008-8830.2017.11.016

论著·实验研究

冬凌草甲素通过下调Brg1表达抑制Jurkat细胞生长

叶珍珍¹, 薛飞龙¹, 丁文评², 孔祥³, 沈伊娜¹

作者信息 +

Oridonin inhibits proliferation of Jurkat cells via the down-regulation of Brg1

YE Zhen-Zhen¹, XUE Fei-Long¹, DING Wen-Ping², KONG Xiang³, SHEN Yi-Na¹

Author information +

文章历史 +

摘要

目的探讨冬凌草甲素对人急性T淋巴细胞白血病Jurkat细胞的作用及其机制。方法体外培养Jurkat细胞株，用不同浓度冬凌草甲素（0、1.25、2.5、5和10 μmol/L）作用Jurkat细胞不同时间（24、48、72 h），MTT实验观察细胞增殖情况，荧光显微镜观察不同浓度冬凌草甲素处理Jurkat细胞12 h后的细胞核形态变化。采用Western blot半定量法检测不同浓度冬凌草甲素作用Jurkat细胞24 h，以及5 μmol/L冬凌草甲素处理Jurkat细胞不同时间（0、2、6、12和24 h）后的Brg1、P53和C-myc蛋白表达。用siRNA沉默Jurkat细胞的Brg1，观察其对P53、C-myc蛋白表达以及对增殖的影响。结果与无处理组相比，冬凌草甲素对Jurkat细胞增殖有抑制作用（P < 0.05），且呈浓度和时间依赖性；荧光显微镜下发现，冬凌草甲素处理后的Jurkat细胞出现细胞核浓集、固缩等典型凋亡形态变化。与无处理组相比，5 μmol/L冬凌草甲素作用下，Jurkat细胞的Brg1和C-myc表达下降，而P53表达上升。siRNA沉默Jurkat细胞的Brg1后，P53表达升高，C-myc蛋白表达下降，细胞生长明显受抑（P < 0.05）。结论冬凌草甲素对Jurkat细胞生长具有抑制作用，其机制可能通过影响Brg1信号通路起作用。

Abstract

Objective To investigate the effect of oridonin on the human acute lymphocytic leukemia cell line Jurkat and its mechanism. Methods Jurkat cells were cultured in vitro and treated with various concentrations (0, 1.25, 2.5, 5, and 10 μmol/L) of oridonin for different lengths of time (24, 48, and 72 hours). The proliferation of Jurkat cells was analyzed by MTT assay. The changes in nuclear morphology were evaluated by fluorescence microscopy at 12 hours after treatment with various concentrations of oridonin. The expression levels of Brg1, P53, and C-myc were determined by semi-quantitative Western blot in Jurkat cells treated with various concentrations of oridonin for 24 hours or 5 μmol/L oridonin for various lengths of time (0, 2, 6, 12, and 24 hours). The expression levels of P53 and C-myc and proliferation of Jurkat cells were evaluated after Brg1 expression was knocked down by Brg1-specific siRNA. Results Compared with the control group, the proliferation of oridonin-treated Jurkat cells was significantly inhibited in a concentration-and time-dependent manner (P < 0.05). According to the florescence microscopic analysis, oridonin treatment led to nuclear pyknosis in Jurkat cells. Compared with the control group, Jurkat cells treated with 5 μmol/L oridonin had reduced expression of Brg1 and C-myc but elevated expression of P53. Brg1 knock-down led to a significant reduction in proliferation of Jurkat cells (P < 0.05), up-regulated expression of P53, and down-regulated expression of C-myc. Conclusions Oridonin can inhibit the proliferation of Jurkat cells, probably via the Brg1 signaling pathway.

导出引用

叶珍珍, 薛飞龙, 丁文评, 孔祥, 沈伊娜. 冬凌草甲素通过下调Brg1表达抑制Jurkat细胞生长[J]. 中国当代儿科杂志. 2017, 19(11): 1208-1212 https://doi.org/10.7499/j.issn.1008-8830.2017.11.016

YE Zhen-Zhen, XUE Fei-Long, DING Wen-Ping, KONG Xiang, SHEN Yi-Na. Oridonin inhibits proliferation of Jurkat cells via the down-regulation of Brg1[J]. Chinese Journal of Contemporary Pediatrics. 2017, 19(11): 1208-1212 https://doi.org/10.7499/j.issn.1008-8830.2017.11.016

参考文献

[1] Siegel RL, Miller KD, Jemal A. Cancer Statistics, 2017[J]. CA Cancer J Clin, 2017, 67(1):7-30.
[2] 林愈灯, 宗飒. 儿童恶性血液肿瘤性疾病诊疗进展[J]. 循证医学, 2017, 17(1):25-28.
[3] Zhao Z, Chen Y. Oridonin, a promising antitumor natural product in the chemotherapy of hematological malignancies[J]. Curr Pharm Biotechnol, 2014, 15(11):1083-1092.
[4] Ding Y, Ding C, Ye N, et al. Discovery and development of natural product oridonin-inspired anticancer agents[J]. Eur J Med Chem, 2016, 122:102-117.
[5] 张晓芬, 周丽. 冬凌草甲素及联合柔红霉素对Jurkat细胞增殖抑制作用的观察[J]. 中华肿瘤防治杂志, 2012, 19(16):1227-1230.
[6] Guo Y, Shan Q, Gong Y, et al. Oridonin in combination with imatinib exerts synergetic anti-leukemia effect in Ph⁺ acute lymphoblastic leukemia cells in vitro by inhibiting activation of LYN/mTOR signaling pathway[J]. Cancer Biol Ther, 2012, 13(13):1244-1254.
[7] Shi J, Whyte WA, Zepeda-Mendoza CJ, et al. Role of SWI/SNF in acute leukemia maintenance and enhancer-mediated Myc regulation[J]. Genes Dev, 2013, 27(24):2648-2662.
[8] 王慧慧, 文飞球. 儿童急性淋巴细胞白血病治疗进展[J]. 中国实用儿科杂志, 2012, 27(10):791-794.
[9] 马晶晶, 陈月, 于亮. 急性淋巴细胞白血病耐药机制的研究进展[J]. 中国实验血液学杂志, 2016, 24(1):261-265.
[10] Tian W, Chen SY. Recent advances in the molecular basis of anti-neoplastic mechanisms of oridonin[J]. Chin J Integr Med, 2013, 19(4):315-320.
[11] Yao Z, Xie F, Li M, et al. Oridonin induces autophagy via inhibition of glucose metabolism in p53-mutated colorectal cancer cells[J]. Cell Death Dis, 2017, 8(2):e2633.
[12] Xu S, Yao H, Luo S, et al. A novel potent anticancer compound optimized from a natural oridonin scaffold induces apoptosis and cell cycle arrest through the mitochondrial pathway[J]. J Med Chem, 2017, 60(4):1449-1468.
[13] 张颖, 郑燕芳. BRG1在非小细胞肺癌中的作用研究进展[J]. 广东医学, 2016, 37(21):3283-3286.
[14] Wu Q, Lian JB, Stein JL, et al. The BRG1 ATPase of human SWI/SNF chromatin remodeling enzymes as a driver of cancer[J]. Epigenomics, 2017, 9(6):919-931.
[15] Weissman B, Knudsen KE. Hijacking the chromatin remodeling machinery:impact of SWI/SNF perturbations in cancer[J]. Cancer Res, 2009, 69(21):8223-8230.
[16] Lu P, Roberts CW. The SWI/SNF tumor suppressor complex:Regulation of promoter nucleosomes and beyond[J]. Nucleus, 2013, 4(5):374-378.
[17] Bai J, Mei PJ, Liu H, et al. BRG1 expression is increased in human glioma and controls glioma cell proliferation, migration and invasion in vitro[J]. J Cancer Res Clin Oncol, 2012, 138(6):991-998.
[18] Bai J, Mei P, Zhang C, et al. BRG1 is a prognostic marker and potential therapeutic target in human breast cancer[J]. PLoS One, 2013, 8(3):e59772.
[19] Naidu SR, Love IM, Imbalzano AN, et al. The SWI/SNF chromatin remodeling subunit BRG1 is a critical regulator of p53 necessary for proliferation of malignant cells[J]. Oncogene, 2009, 28(27):2492-2501.