野黄芩苷对人脑胶质瘤U87细胞的增殖抑制作用及其机制

doi:10.13481/j.1671-587x.20180302

摘要/Abstract

摘要： 目的：探讨野黄芩苷（SCU）对人脑胶质瘤U87细胞的增殖抑制作用，并初步阐明其作用机制。方法：培养U87细胞，采用不同浓度（20、40和80 mg·L^-1） SCU处理U87细胞（野黄芩苷组），同时设空白对照组。MTT法检测各组U87细胞存活率，倒置显微镜观察U87细胞的形态表现，Annexin Ⅴ/PI双染检测各组U87细胞凋亡率，Western blotting法检测各组U87细胞中bax和bcl-2蛋白表达水平。结果：MTT法，与空白对照组比较，20、40和80 mg·L^-1SCU组细胞存活率均明显降低（P<0.01）。倒置显微镜观察，与空白对照组比较，80 mg·L^-1SCU组细胞数明显减少，细胞体积变小，细胞间距变大；细胞出现皱缩、脱落，贴壁细胞轮廓模糊。Annexin Ⅴ/PI双染，与空白对照组比较，80 mg·L^-1SCU组细胞晚期凋亡率和总凋亡率均明显升高（P<0.01）。Western blotting法，与空白对照组比较，20、40和80 mg·L^-1 SCU组U87细胞中bcl-2蛋白水平降低，80 mg·L^-1SCU组差异有统计学意义（P<0.01）；与空白对照组比较，80 mg·L^-1 SCU组U87细胞中bax/bcl-2比值升高（P<0.01）。结论：SCU通过上调U87细胞中bax/bcl-2比值诱导细胞凋亡从而抑制肿瘤细胞的生长。

关键词: 细胞凋亡, 野黄芩苷, 胶质瘤细胞, 细胞增殖, bcl-2, bax

Abstract: Objective: To explore the inhibitory effect of scutellarin(SCU) on the proliferation of human glioma U87 cells,and to clarify its mechanism. Methods: The U87 cells were cultured and divided into blank control group and 20,40,80 mg·L^-1 SCU groups,the surival rates of U87 cells in various groups were examined by MTT assay; the morphology of U87 cells in various groups were observed by inverted microscope;the apoptotic rates of U87 cells were examined by Annexin Ⅴ/PI staining; the expression levels of bax and bcl-2 proteins in the U87 cells in various groups were detected by Western blotting method. Results: The MTT results showed that compared with blank control group,the survival rates of U87 cells in 20,40,and 80 mg·L^-1 SCU groups were decreased(P<0.01).The inverted microscope observation results showed that compared with blank control group,the number of U87 cells in 80 mg·L^-1 SCU group was significantly decreased,and the cell volume was smaller;the U87 cells were shrinkage,shedding,and the outline of adherent cells was fuzzy.The results of AnnexinⅤ/PI staining showed that the late and total apoptotic rates of U87 cells in 80 mg·L^-1 SCU group were increased compared with blank control group (P<0.01).The Western blotting results showed that compared with blank control group,the expression levels of bcl-2 protein in the U87 cells in 20,40 and 80 mg·L^-1 SCU groups were decreased,especially in 80 mg·L^-1 SCU group(P<0.01).Compared with blank control group,the ratio of bax/bcl-2 in the U87 cells in 80 mg·L^-1 SCU group was significantly increased(P<0.01). Conclusion: SCU can induce the apoptosis and suppress the tumor growth by upregulating the bax/bcl-2 ratio in the U87 cells.

Key words: bcl-2, cell proliferation, apoptosis, scutellarin, glioma, bax

中图分类号:

R181.2

吕鹏, 宁明杰, 邵玉, 张璐妮, 唐英, 王南博, 陈飞儿, 齐玲. 野黄芩苷对人脑胶质瘤U87细胞的增殖抑制作用及其机制[J]. 吉林大学学报(医学版), 2018, 44(03): 466-470.

LYU Peng, NING Mingjie, SHAO Yu, ZHANG Luni, TANG Ying, WANG Nanbo, CHEN Feier, QI Ling. Inhibitory effect of scutellarin on proliferation of human glioma U87 cellsand its mechanism[J]. Journal of Jilin University Medicine Edition, 2018, 44(03): 466-470.

参考文献

[1] Qi L,Ding L,Wang S,et al.A network meta-analysis:the overall and progression-free survival of glioma patients treated by different chemotherapeutic interventions combined with radiation therapy (RT)[J].Oncotarget,2016,7(42):69002-69013.
[2] 齐玲,杨阳,刘玉翠,等.双氢青蒿素抑制神经母细胞瘤细胞生长的作用[J].吉林大学学报:医学版, 2016,42 (2):266-270.
[3] Jiang NH,Zhang GH,Zhang JJ,et al. Analysis of the transcriptome of Erigeron breviscapus uncovers putative scutellarin and chlorogenic acidsbiosynthetic genes and genetic markers[J].PLoS One,2014,9(6):e100357.
[4] Zhu PT,Mao M,Liu ZG,et al.Scutellarin suppresses human colorectal cancer metastasis and angiogenesis by targeting ephrinb[J].Am J Transl Res, 2017,9(11):5094-5104.
[5] 葛志华,张颖,李俊玫,等.野黄芩苷对人舌鳞癌Tca8113细胞Bcl-2、Bax表达的影响[J].实用口腔医学杂志,2014,30(5):649-652.
[6] 王行天,孙立新,杨宇,等.野黄芩苷对人舌鳞癌细胞的增殖移植和Bcl-2蛋白表达的影响[J].实用口腔医学杂志,2013,29(1):83-85.
[7] You L,Zhu H,Wang C,et al.Scutellarin inhibits Hela cell growth and glycolysis by inhibiting the activity of pyruvate kinase M2[J].Bioorg Med Chem Lett,2017,27(24):5404-5408.
[8] Hou L,Chen L,Fang L.Scutellarin inhibits proliferation,invasion,and tumorigenicity in human breast cancer cells by regulating HIPPO-YAP signaling pathway[J].Med Sci Monit,2017,23:5130-5138.
[9] Zhang L,Sun S,Li W,et al.Effect of Scutellarin inhibits collagen-induced arthritis through TLR4/NF-κB-mediated inflammation[J].Mol Med Rep,2017,16(4):5555-5560.
[10] Wesseling J,Hartog H, Horlings H,et al.Different effects of insulin-like growth factor-1 receptor expression on prognosis of estrogen receptor positive versus triple-negative invasive ductal breast carcinoma[J].J Clin Oncol,2009,27(15 suppl):3546.
[11] Qi L,Bellail AC,Rossi MR,et al.Heterogeneity of primary glioblastoma cells in the expression of caspase-8 and the response to TRAIL-induced apoptosis[J].Apoptosis,2011,16(11):1150-1164.
[12] 颜承,徐冠玲,谢梦,等.野黄芩苷和野黄芩素制备研究进展[J].中成药,2015,37(8):1785-1790.
[13] Han T,Li J,Xue J,et al.Scutellarin derivatives as apoptosis inducers:Design,synthesis and biological evaluation[J].Eur J Med Chem,2017,135:270-281.
[14] Yang N,Zhao Y,Wang Z,et al.Scutellarin suppresses growth and causes apoptosis of human colorectal cancer cells by regulating the p53 pathway[J].Mol Med Rep,2017,15(2):929-935.
[15] Wang W,Ma X,Han J,et al.Neuroprotective effect of scutellarin on ischemic cerebral injury by down-regulating the expression of angiotensin-converting enzyme and AT1receptor[J].PLoS One,2016,11(1):e0146197.
[16] Niu C,Sheng Y,Yang R,et al.Scutellarin protects against the liver injury induced by diosbulbin B in mice and its mechanism[J].J Ethnopharmacol,2015,164:301-308.
[17] 郭晓菲,高树建,闫朝岐,等.野黄芩苷抑制人乳腺癌细胞系MCF-7的增殖并上调miRNA表达[J].现代肿瘤医学,2016,24(2):197-200.
[18] Qi L,Yu HQ,Li YQ,et al.Schidandrin B kills tumor cells by initiating apoptosis in glioma SHG-44 cells[J].Chin J Integr Med,2016.doi:10.1007/s11655-015-2406-9.
[19] Li H,Huang D,Gao Z,et al.Scutellarin inhibits the growth and invasion of human tongue squamous carcinoma through the inhibition of matrix metalloproteinase-2 and-9 and avβ6 integrin[J].Int J Oncol,2013, 42(5):1674-1681.
[20] 邵玉,张璐妮,宁明杰,等.野黄芩苷诱导肿瘤细胞凋亡作用机制的研究进展[J].吉林医药学院学报,2017,38(4):298-301.
[21] Qi L,Bellail AC,Rossi MR, et al.Heterogeneity of primary glioblastoma cells in the expression of caspase-8 and the response to TRAIL-induced apoptosis[J].Apoptosis,2011,16(11):1150-1164.
[22] 张瑞,李成云,苏莉,等.原花青素对硫酸镍染毒大鼠肝脏细胞Capase-3、Bcl-2和Bax蛋白表达的影响[J].工业卫生与职业病,2013,39(5):276-279.
[23] 王双双,孔根现,杨升华,等.骨髓间充质干细胞拮抗氧化型低密度脂蛋白致人脐静脉内皮细胞凋亡[J].中国动脉硬化杂志,2013,21(2):105-108.
[24] 刘志军,周海存,陈小兵,等.人脑胶质瘤组织中 CPEB4 的表达及与细胞迁移和侵袭的关系[J].郑州大学学报:医学版,2016,51(5):622-626.