白藜芦醇通过调控SIRT1抑制卵巢癌细胞生长及Wnt信号通路的研究
|
摘要
目的研究白藜芦醇对卵巢癌A2780细胞生长的影响,并探讨其作用机制。方法运用MTT法检测白藜芦醇(50、100、200、400μmol/L)对A2780细胞的抑制率;白藜芦醇+pc DNA3.1组(转染pc DNA3.1)、白藜芦醇+pc DNA3.1-SIRT1组(转染pc DNA3.1-SIRT1)均用脂质体法转染,加入白藜芦醇(200μmol/L)处理;采用实时荧光定量PCR(q RT-PCR)法检测细胞中信息调节因子1(SIRT1)、β-catenin、c-Myc的m RNA水平;Western blotting法检测细胞中SIRT1、β-catenin、c-Myc蛋白表达;流式细胞术检测细胞凋亡率。结果与对照组比较,白藜芦醇(50、100、200、400μmol/L)处理组细胞增殖抑制率、凋亡率均显著升高(P<0.05),且白藜芦醇(200μmol/L)组细胞中SIRT1的m RNA和蛋白水平均显著降低(P<0.05),可失活Wnt信号通路;过表达SIRT1可逆转白藜芦醇对A2780细胞增殖及Wnt信号通路的失活作用。结论白藜芦醇可调控SIRT1抑制卵巢癌细胞增殖,促进凋亡,其促凋亡机制可能与失活Wnt信号通路有关。
Objective To study the effects of resveratrol on cell proliferation and apoptosis of A2780 cells, and explore its mechanism. Methods MTT assay was used to detect inhibition rate of A2780 cells with resveratrol(50, 100, 200, and 400 μmol/L); Resveratrol + pc DNA3.1 group(transfected pcDNA3.1), and resveratrol + pcDNA3.1-SIRT1 group(transfected pcDNA3.1-SIRT1) were transfected into A2780 cells by liposome method, and treated with resveratrol(200 μmol/L); The mRNA levels of SIRT1, β-catenin, and c-Myc were detected by qRT-PCR. The protein expression of SIRT1, β-catenin, and c-Myc were detected by Western blotting. The apoptosis rate of each group was detected by flow cytometry. Results Compared with the blank control group, the inhibition rate and apoptosis rate were significantly increased(P < 0.05) in A2780 cells with resveratrol(50, 100, 200, and 400 μmol/L), and the mRNA and protein levels of SIRT1 were significantly decreased(P < 0.05) in the resveratrol(200 μmol/L) group, which could inactivate the Wnt signaling pathway. Overexpression SIRT1 reversed the inhibitory effect of resveratrol on A2780 cell proliferation and the inactivation of Wnt signaling pathway. Conclusion Resveratrol can inhibit ovarian cancer cell proliferation and promote apoptosis in ovarian cancer cells by regulating SIRT1, and its pro-apoptotic mechanism may be relate to inactivated Wnt signaling pathway, which will provide a basis for resveratrol treatment of ovarian cancer.
Objective To study the effects of resveratrol on cell proliferation and apoptosis of A2780 cells, and explore its mechanism. Methods MTT assay was used to detect inhibition rate of A2780 cells with resveratrol(50, 100, 200, and 400 μmol/L); Resveratrol + pc DNA3.1 group(transfected pcDNA3.1), and resveratrol + pcDNA3.1-SIRT1 group(transfected pcDNA3.1-SIRT1) were transfected into A2780 cells by liposome method, and treated with resveratrol(200 μmol/L); The mRNA levels of SIRT1, β-catenin, and c-Myc were detected by qRT-PCR. The protein expression of SIRT1, β-catenin, and c-Myc were detected by Western blotting. The apoptosis rate of each group was detected by flow cytometry. Results Compared with the blank control group, the inhibition rate and apoptosis rate were significantly increased(P < 0.05) in A2780 cells with resveratrol(50, 100, 200, and 400 μmol/L), and the mRNA and protein levels of SIRT1 were significantly decreased(P < 0.05) in the resveratrol(200 μmol/L) group, which could inactivate the Wnt signaling pathway. Overexpression SIRT1 reversed the inhibitory effect of resveratrol on A2780 cell proliferation and the inactivation of Wnt signaling pathway. Conclusion Resveratrol can inhibit ovarian cancer cell proliferation and promote apoptosis in ovarian cancer cells by regulating SIRT1, and its pro-apoptotic mechanism may be relate to inactivated Wnt signaling pathway, which will provide a basis for resveratrol treatment of ovarian cancer.
引文
[1]孙玮,李红雨,臧星卉,等.上皮性卵巢癌组织中Slug和E-cadherin的表达[J].郑州大学学报:医学版,2018,53(3):374-378.
[2]BaczkóI,Light P E.Resveratrol and derivatives for the treatment of atrial fibrillation[J].Ann N Y Acad Sci,2015,1348(1):68-74.
[3]Ozkan O V,Yuzbasioglu M F,Ciralik H,et al.Resveratrol,a natural antioxidant,attenuates intestinal ischemia/reperfusion injury in rats[J].Tohoku J Exp Med,2009,218(3):251-258.
[4]Dong W,Li N,Gao D,et al.Resveratrol attenuates ischemic brain damage in the delayed phase after stroke and induces mesenger RNA and protein express foe angiogenic factors[J].J Vasc Surg,2008,48(3):709-714.
[5]Simic P,Williams E O,Bell E L,et al.SIRT1 suppresses the epithelial-to-mesenchymal transition in cancer metastasis and organ fibrosis[J].Cell Rep,2013,3(4):1175-1186.
[6]Yang H,Bi Y,Xue L,et al.Multifaceted modulation of SIRT1 in cancer and inflammation[J].Crit Rev Oncog,2015,20(1/2):49-64.
[7]Simmons G E J,Pruitt W M,Pruitt K.Diverse roles of SIRT1 in cancer biology and lipid metabolism[J].Int JMol Sci,2015,16(1):950-965.
[8]史贵月,毕芳芳,杨清.BRCA1、SIRT1与卵巢癌关系的研究进展[J].医学综述,2017,23(9):1757-1760.
[9]Mei Y Z,Liu R X,Wang D P,et al.Biocatalysis and biotransformation of resveratrol in microorganisms[J].Biotechnol Lett,2015,37(1):9-18.
[10]刘顺,李赫宇,赵玲.白藜芦醇降血尿酸、抗炎作用研究进展[J].药物评价研究,2016,29(2):304-307.
[11]王欣欣,马珊珊,孟楠,等.h UC-MSCs和白藜芦醇对AD小鼠学习记忆能力及脑内SIRT1信号通路的影响[J].郑州大学学报:医学版,2016,51(5):576-579.
[12]伍莉,田景鸣.白藜芦醇抑制人卵巢癌细胞株A2780细胞增殖及凋亡诱导作用[J].中国中医药现代远程教育,2008(6):596-597.
[13]Cerutti R,Pirinen E,Lamperti C,et al.NAD(+)-dependent activation of Sirt1 corrects the phenotype in a mouse model of mitochondrial disease[J].Cell Metab,2014,19(6):1042-1049.
[14]Wang Y,Li D,Ma G,et al.Increases in peripheral SIRT1:A new biological characteristic of asthma[J].Respirology,2015,20(7):1066-1072.
[15]路富林,余琦,魏明.运动预处理对大鼠心脏缺血再灌注损伤的保护作用[J].郑州大学学报:医学版,2018,53(3):323-327.
[16]谢可炜,魏凯,顾明颖,等.白藜芦醇通过Sirtuin1激活足细胞Notch1信号[J].中华肾脏病杂志,2015,31(4):296-302.
[17]孙军营,林健静,栾昕宇,等.沉默信息调节因子1基因对软骨细胞胞外基质影响的体外研究[J].中华骨与关节外科杂志,2018,11(6):448-453.
[18]Cruciat C M,Niehrs C.Secreted and transmembrane wnt inhibitors and activators[J].Cold Spring Harb Perspect Biol,2013,5(3):a015081.
[19]Polakis P.Wnt signaling in cancer[J].Cold Spring Harb Perspect Biol,2012,4(5):a008052.
[20]郭玉霞,马利国,陈递林,等.Wnt信号通路成分相关因子在卵巢癌中的表达以及临床预后价值[J].山西医药杂志,2018,47(13):1518-1521.
[21]Cao P L,Zhao S,Sun Z G,et al.BRMS1L suppresses ovarian cancer metastasis via inhibition of theβ-catenin-wnt pathway[J].Exp Cell Res,2018,371(1):214-221.
[22]Hui L,Li H Y,Lu G,et al.Low dose of bisphenol a modulates ovarian cancer gene expression profile and promotes epithelial to mesenchymal transition via canonical Wnt pathway[J].Toxicol Sci,2018,164(2):527-538.
[23]Yu F,Yuan Y,Li D,et al.The effect of lentivirus-mediated SIRT1 gene knockdown in the ATDC5 cell line via inhibition of the Wnt signaling pathway[J].Cell Signal,2018,doi:10.1016/j.cellsig.2018.09.016.
[2]BaczkóI,Light P E.Resveratrol and derivatives for the treatment of atrial fibrillation[J].Ann N Y Acad Sci,2015,1348(1):68-74.
[3]Ozkan O V,Yuzbasioglu M F,Ciralik H,et al.Resveratrol,a natural antioxidant,attenuates intestinal ischemia/reperfusion injury in rats[J].Tohoku J Exp Med,2009,218(3):251-258.
[4]Dong W,Li N,Gao D,et al.Resveratrol attenuates ischemic brain damage in the delayed phase after stroke and induces mesenger RNA and protein express foe angiogenic factors[J].J Vasc Surg,2008,48(3):709-714.
[5]Simic P,Williams E O,Bell E L,et al.SIRT1 suppresses the epithelial-to-mesenchymal transition in cancer metastasis and organ fibrosis[J].Cell Rep,2013,3(4):1175-1186.
[6]Yang H,Bi Y,Xue L,et al.Multifaceted modulation of SIRT1 in cancer and inflammation[J].Crit Rev Oncog,2015,20(1/2):49-64.
[7]Simmons G E J,Pruitt W M,Pruitt K.Diverse roles of SIRT1 in cancer biology and lipid metabolism[J].Int JMol Sci,2015,16(1):950-965.
[8]史贵月,毕芳芳,杨清.BRCA1、SIRT1与卵巢癌关系的研究进展[J].医学综述,2017,23(9):1757-1760.
[9]Mei Y Z,Liu R X,Wang D P,et al.Biocatalysis and biotransformation of resveratrol in microorganisms[J].Biotechnol Lett,2015,37(1):9-18.
[10]刘顺,李赫宇,赵玲.白藜芦醇降血尿酸、抗炎作用研究进展[J].药物评价研究,2016,29(2):304-307.
[11]王欣欣,马珊珊,孟楠,等.h UC-MSCs和白藜芦醇对AD小鼠学习记忆能力及脑内SIRT1信号通路的影响[J].郑州大学学报:医学版,2016,51(5):576-579.
[12]伍莉,田景鸣.白藜芦醇抑制人卵巢癌细胞株A2780细胞增殖及凋亡诱导作用[J].中国中医药现代远程教育,2008(6):596-597.
[13]Cerutti R,Pirinen E,Lamperti C,et al.NAD(+)-dependent activation of Sirt1 corrects the phenotype in a mouse model of mitochondrial disease[J].Cell Metab,2014,19(6):1042-1049.
[14]Wang Y,Li D,Ma G,et al.Increases in peripheral SIRT1:A new biological characteristic of asthma[J].Respirology,2015,20(7):1066-1072.
[15]路富林,余琦,魏明.运动预处理对大鼠心脏缺血再灌注损伤的保护作用[J].郑州大学学报:医学版,2018,53(3):323-327.
[16]谢可炜,魏凯,顾明颖,等.白藜芦醇通过Sirtuin1激活足细胞Notch1信号[J].中华肾脏病杂志,2015,31(4):296-302.
[17]孙军营,林健静,栾昕宇,等.沉默信息调节因子1基因对软骨细胞胞外基质影响的体外研究[J].中华骨与关节外科杂志,2018,11(6):448-453.
[18]Cruciat C M,Niehrs C.Secreted and transmembrane wnt inhibitors and activators[J].Cold Spring Harb Perspect Biol,2013,5(3):a015081.
[19]Polakis P.Wnt signaling in cancer[J].Cold Spring Harb Perspect Biol,2012,4(5):a008052.
[20]郭玉霞,马利国,陈递林,等.Wnt信号通路成分相关因子在卵巢癌中的表达以及临床预后价值[J].山西医药杂志,2018,47(13):1518-1521.
[21]Cao P L,Zhao S,Sun Z G,et al.BRMS1L suppresses ovarian cancer metastasis via inhibition of theβ-catenin-wnt pathway[J].Exp Cell Res,2018,371(1):214-221.
[22]Hui L,Li H Y,Lu G,et al.Low dose of bisphenol a modulates ovarian cancer gene expression profile and promotes epithelial to mesenchymal transition via canonical Wnt pathway[J].Toxicol Sci,2018,164(2):527-538.
[23]Yu F,Yuan Y,Li D,et al.The effect of lentivirus-mediated SIRT1 gene knockdown in the ATDC5 cell line via inhibition of the Wnt signaling pathway[J].Cell Signal,2018,doi:10.1016/j.cellsig.2018.09.016.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |