蓝萼甲素对三阴性乳腺癌MDA-MB-231细胞增殖及细胞周期的影响
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摘要
目的研究蓝萼甲素对三阴性乳腺癌MDA-MB-231细胞增殖及细胞周期的影响及其作用机制。方法采用MTT法检测蓝萼甲素对MDA-MB-231细胞增殖的影响;流式细胞仪检测细胞周期;Western blotting法检测细胞中cyclin B1、cyclin D1、细胞周期素依赖激酶2(CDK2)、CDK4、p53、p21、p27、组蛋白赖氨酸特异性去甲基化酶1(LSD1)、组蛋白H3第4位赖氨酸二甲基化(H3K4me2)、组蛋白H3第9位赖氨酸二甲基化(H3K9me2)蛋白表达水平。结果蓝萼甲素能显著抑制MDA-MB-231细胞的增殖,呈剂量和时间依赖性;提高G2/M期细胞比例;上调p53、p21、p27、H3K4me2、H3K9me2蛋白表达水平,下调cyclin B1、cyclin D1、CDK2、CDK4、LSD1蛋白表达水平。结论蓝萼甲素抑制MDA-MB-231细胞增殖,阻滞MDA-MB-231细胞周期于G2/M期,其机制可能与激活p53的表达及调控组蛋白的甲基化作用有关。
Objective To investigate the effect of glaucocalyxin A on proliferation and cell cycle of triple-negative breast cancer MDA-MB-231 cells and its mechanism. Methods The proliferation inhibition rates of MDA-MB-231 cells were measured by MTT assay. The cell cycle was analyzed by flow cytometry, and the expression of the protein cyclin B1, cyclin D1, CDK2, CDK4, p53, p21,p27, LSD1, H3 K4 me2, and H3 K9 me2 was detected by Western blotting. Results Growth of MDA-MB-231 cells was significantly inhibited by glaucocalyxin A in a dose-dependent and time-dependent manner. Flow cytometric analysis indicated that the percentage of MDA-MB-231 cells at G2/M phase was increased significantly. As the results of Western blotting, the protein expression levels of p53, p21, p27, H3 K4 me2, and H3 K9 me2 in MDA-MB-231 cells were increased, while that of cyclin B1, cyclin D1, CDK2, CDK4, and LSD1 were decreased after treated with glaucocalyxin A. Conclusion Glaucocalyxin A could inhibit the proliferation of MDA-MB-231 cells and induce cell cycle arrest at the G2/M phase, and the mechanism may be related to the activation of p53 protein expression and the regulation of histone methylation.
Objective To investigate the effect of glaucocalyxin A on proliferation and cell cycle of triple-negative breast cancer MDA-MB-231 cells and its mechanism. Methods The proliferation inhibition rates of MDA-MB-231 cells were measured by MTT assay. The cell cycle was analyzed by flow cytometry, and the expression of the protein cyclin B1, cyclin D1, CDK2, CDK4, p53, p21,p27, LSD1, H3 K4 me2, and H3 K9 me2 was detected by Western blotting. Results Growth of MDA-MB-231 cells was significantly inhibited by glaucocalyxin A in a dose-dependent and time-dependent manner. Flow cytometric analysis indicated that the percentage of MDA-MB-231 cells at G2/M phase was increased significantly. As the results of Western blotting, the protein expression levels of p53, p21, p27, H3 K4 me2, and H3 K9 me2 in MDA-MB-231 cells were increased, while that of cyclin B1, cyclin D1, CDK2, CDK4, and LSD1 were decreased after treated with glaucocalyxin A. Conclusion Glaucocalyxin A could inhibit the proliferation of MDA-MB-231 cells and induce cell cycle arrest at the G2/M phase, and the mechanism may be related to the activation of p53 protein expression and the regulation of histone methylation.
引文
[1]Harbeck N,Gnant M.Breast cancer[J].Lancet,2017,389(10074):1134-1150.
[2]Gluz O,Liedtke C,Gottschalk N,et al.Triple-negative breast cancer-current status and future directions[J].Ann Oncol,2009,20(12):1913-1927.
[3]李甜,周钱梅,张卫红.益气扶正复方联合依维莫司对三阴性乳腺癌细胞株MDA-MB-468的增殖抑制作用及其对Akt/mTOR信号通路的影响[J].世界科学技术-中医药现代化,2018,20(1):37-43.
[4]李盛建,王莹,王强利.月腺大戟素A抗乳腺癌活性[J].第二军医大学学报,2018,39(7):765-769.
[5]O’Reilly E A,Gubbins L,Sharma S,et al.The fate of chemoresistance in triple negative breast cancer(TNBC)[J].BBA Clin,2015,doi:10.1016/j.bbacli.2015.03.003.
[6]迟晴晴,王强,钟延强,等.蓝萼甲素的药理活性及其机制和毒理作用的研究进展[J].药学实践杂志,34(2):124-128.
[7]Po L S,Chen Z Y,Tsang D S,et al.Baicalein and genistein display differential actions on estrogen receptor(ER)transactivation and apoptosis in MCF-7 cells[J].Cancer Lett,2002,187(12):33-40.
[8]李娟,胡永华.葛根素对人小细胞肺癌H446细胞周期和相关周期蛋白表达的影响[J].中草药,2008,39(10):1535-1537.
[9]Liu Z,Liu H,Yuan X,et al.Downregulation of Pim-2induces cell cycle arrest in the G0/G1 phase via the p53-non-dependent p21 signaling pathway[J].Oncol Lett,2018,15(4):4079-4086.
[10]Zheng R,Liu Y,Zhang X,et al.miRNA-200C enhances radiosensitivity of esophageal cancer by cell cycle arrest and targeting p21[J].Biomed Pharmacother,2017,doi:10.1016/j.biopha.2017.04.006.
[11]Luo Y,Chen X,Luo L,et al.[6]-Gingerol enhances the radiosensitivity of gastric cancer via G2/M phase arrest and apoptosis induction[J].Oncol Rep,2018,39(5):2252-2260.
[12]赵春玲,宋咏梅,樊飞跃,等.细胞周期蛋白cyclin B1与肿瘤[J].肿瘤,2007,27(4):322-326.
[13]葛亚楠,朱欢欢,韩涛,等.细胞周期蛋白在三阴型和激素依赖型乳腺癌中的差异表达及临床意义[J].解放军医药杂志,2016,28(6):27-32.
[14]马英玉.LSD1及与肿瘤关系研究进展[J].中国肿瘤,2010,19(9):599-603.
[15]Metzger E,Wissmann M,Yin N,et al.LSD1demethylates repressive histone marks to promote androgen-receptor-dependent transcription[J],Nature,2005,437(7057):436-439.
[16]Yokoyama A,Igarashi K,Sato T,et al.Identification of myelin transcription factor 1(MyT1)as a subunit of the neural cell type-specific lysine-specific demethylase 1(LSD1)complex[J].J Biol Chem,2014,289(26):18152-18162.
[17]Laurent B,Ruitu L,Murn J,et al.A specific LSD1/KDM1A isoform regulates neuronal differentiation through H3K9 demethylation[J].Mol Cell,2015,57(6):957-970.
[18]李航,张劲帆,陈宇,等.LSD1在乳腺癌中的表达及意义[J].莆田学院学报,2016,23(5):27-30.
[2]Gluz O,Liedtke C,Gottschalk N,et al.Triple-negative breast cancer-current status and future directions[J].Ann Oncol,2009,20(12):1913-1927.
[3]李甜,周钱梅,张卫红.益气扶正复方联合依维莫司对三阴性乳腺癌细胞株MDA-MB-468的增殖抑制作用及其对Akt/mTOR信号通路的影响[J].世界科学技术-中医药现代化,2018,20(1):37-43.
[4]李盛建,王莹,王强利.月腺大戟素A抗乳腺癌活性[J].第二军医大学学报,2018,39(7):765-769.
[5]O’Reilly E A,Gubbins L,Sharma S,et al.The fate of chemoresistance in triple negative breast cancer(TNBC)[J].BBA Clin,2015,doi:10.1016/j.bbacli.2015.03.003.
[6]迟晴晴,王强,钟延强,等.蓝萼甲素的药理活性及其机制和毒理作用的研究进展[J].药学实践杂志,34(2):124-128.
[7]Po L S,Chen Z Y,Tsang D S,et al.Baicalein and genistein display differential actions on estrogen receptor(ER)transactivation and apoptosis in MCF-7 cells[J].Cancer Lett,2002,187(12):33-40.
[8]李娟,胡永华.葛根素对人小细胞肺癌H446细胞周期和相关周期蛋白表达的影响[J].中草药,2008,39(10):1535-1537.
[9]Liu Z,Liu H,Yuan X,et al.Downregulation of Pim-2induces cell cycle arrest in the G0/G1 phase via the p53-non-dependent p21 signaling pathway[J].Oncol Lett,2018,15(4):4079-4086.
[10]Zheng R,Liu Y,Zhang X,et al.miRNA-200C enhances radiosensitivity of esophageal cancer by cell cycle arrest and targeting p21[J].Biomed Pharmacother,2017,doi:10.1016/j.biopha.2017.04.006.
[11]Luo Y,Chen X,Luo L,et al.[6]-Gingerol enhances the radiosensitivity of gastric cancer via G2/M phase arrest and apoptosis induction[J].Oncol Rep,2018,39(5):2252-2260.
[12]赵春玲,宋咏梅,樊飞跃,等.细胞周期蛋白cyclin B1与肿瘤[J].肿瘤,2007,27(4):322-326.
[13]葛亚楠,朱欢欢,韩涛,等.细胞周期蛋白在三阴型和激素依赖型乳腺癌中的差异表达及临床意义[J].解放军医药杂志,2016,28(6):27-32.
[14]马英玉.LSD1及与肿瘤关系研究进展[J].中国肿瘤,2010,19(9):599-603.
[15]Metzger E,Wissmann M,Yin N,et al.LSD1demethylates repressive histone marks to promote androgen-receptor-dependent transcription[J],Nature,2005,437(7057):436-439.
[16]Yokoyama A,Igarashi K,Sato T,et al.Identification of myelin transcription factor 1(MyT1)as a subunit of the neural cell type-specific lysine-specific demethylase 1(LSD1)complex[J].J Biol Chem,2014,289(26):18152-18162.
[17]Laurent B,Ruitu L,Murn J,et al.A specific LSD1/KDM1A isoform regulates neuronal differentiation through H3K9 demethylation[J].Mol Cell,2015,57(6):957-970.
[18]李航,张劲帆,陈宇,等.LSD1在乳腺癌中的表达及意义[J].莆田学院学报,2016,23(5):27-30.