莪术醇抗T47D乳腺癌细胞增殖的作用机制研究
x

请在关注微信后,向客服人员索取文件

篇名: 莪术醇抗T47D乳腺癌细胞增殖的作用机制研究
TITLE: Study on the mechanism of curcumol inhibiting the proliferation of breast cancer cells T 47D
摘要: 目的 研究莪术醇抗T47D乳腺癌细胞增殖的作用机制。方法采用MTT法检测不同剂量莪术醇(0、6.25、12.5、25、50、100μg/mL)对T47D乳腺癌细胞增殖的抑制作用。以莪术醇(12.5、25、50、100μg/mL)干预T47D乳腺癌细胞后,采用倒置相差显微镜观察细胞形态;采用流式细胞仪检测细胞周期及活性氧(ROS)水平;采用实时荧光定量聚合酶链反应(qRT-PCR)法检测增殖细胞核抗原(PCNA)、细胞周期调控因子p21和细胞周期蛋白依赖性激酶2(CDK2)的mRNA表达水平;采用Westernblot法检测CDK2、CDK6、细胞周期蛋白D(CyclinD)、PCNA、核转录因子E2-相关因子(Nrf2)、Kelch样ECH关联蛋白1(Keap1)的蛋白表达水平。另将细胞分为2组,其中一组加入不同剂量莪术醇,另外一组加入33μg/mL莪术醇干预6、12、24、48h,根据以上2组结果确定最佳氧化时间和给药浓度后,另设空白对照组、N-乙酰半胱氨酸(NAC)组(单用ROS抗氧化剂NAC)、莪术醇组(单用莪术醇)、莪术醇联合NAC组(采用ROS抗氧化剂NAC预处理,再加入莪术醇),检测细胞周期及ROS荧光强度。结果莪术醇能使T47D乳腺癌细胞增殖抑制率显著升高(P<0.05或P<0.01),并呈一定的剂量和时间依赖趋势。莪术醇能将T47D乳腺癌细胞周期阻滞于G1期,并使ROS水平显著增加(P<0.05或P<0.01);ROS抗氧化剂NAC能够逆转莪术醇的上述诱导作用(P<0.01)。qRT-PCR结果显示,莪术醇能下调PCNA和CDK2mRNA的表达,并上调p21mRNA的表达(P<0.05或P<0.01)。Westernblot结果显示,莪术醇能显著下调Keap1、Nrf2、CDK2、CDK6和CyclinD的蛋白表达(P<0.05或P<0.01);ROS抗氧化剂NAC能够逆转莪术醇对上述蛋白表达的下调作用(P<0.05或P<0.01)。结论莪术醇可能通过诱导氧化应激和细胞周期阻滞,发挥抑制T47D乳腺癌细胞增殖的作用。
ABSTRACT: OBJECTIVE To study the mechanism of curcumol inhibiting the pro liferation of breast cancer cells T 47D. METHODS MTT assay was used to detect the inhibitory effects of different doses of curcumol (0,6.25,12.5,25,50,100 μg/mL)on the proliferation of T 47D cells. After treated with curcumol (12.5,25,50,100 μg/mL),the morphology of T 47D cells was observed by inverted phase contrast microscope. The cell cycle and the levels of reactive oxygen species (ROS)were detected by flow cytometry. Quantitative real-time PCR (qRT-PCR)was used to detect the expressions of proliferating cell nuclear antigen (PCNA),cell cycle regular p 21 and cyclin-dependent kinase 2(CDK2)mRNA. Western blot assay was used to detect the protein expression of CDK 2,CDK6,Cyclin D ,PCNA,nucler transcription factor E 2-related factor (Nrf2)and Kelch-like ECH associated protein 1(Keap1). Breast cancer cells T 47D were divided into 2 groups,one group was given different doses of curcumol ,and another group was given curcumol 33 μg/mL for 6,12,24,48 h. After the optimal oxidation time and administration concentration were determined according to the results of the above two groups ,the blank control group ,N-acetylcysteine(NAC)group(ROS antioxidant NAC alone ),curcumol group (curcumol alone ),curcumol combined with NAC group (pretreatment with ROS antioxidant NAC ,and then adding into curcumol ). Cell cycle and fluorescence intensity of ROS were detected. RESULTS Curcumol could significantly increase the inhibitory rate of the proliferation of T 47D cells (P<0.05 or P<0.01),and showed a certain dose and time dependent trend. Curcumol blocked the , cycle in the G 1 phase and significantly increased the level of ROS (P<0.05 or P<0.01);ROS antioxidant NAC could significantly reverse above inductive effect of curcumol (P< 0.01). qRT-PCR showed that curcumol down-regulated the com expression of PCNA and CDK 2 mRNA and up-regulated the expression of p 21 mRNA(P<0.05 or P<0.01). Western blot assay showed that curcumol significantly down-regulated the edu.cn protein expression of Keap 1,Nrf2,CDK2,CDK6 and Cyclin D(P<0.05,P<0.01);ROS antioxidant NAC could reverse the down-regulation effects of curcumol on the expression of these proteins(P<0.05 or P<0.01). CONCLUSIONS Curcumol may induce oxidative stress and cell arrest in G 1 phase to inhibit the proliferation of T 47D cells.
期刊: 2022年第33卷第05期
作者: 周璐炜,王娟,陈旭
AUTHORS: ZHOU Luwei,WANG Juan,CHEN Xu
关键字: 莪术醇;T47D乳腺癌细胞;细胞周期阻滞;细胞增殖;氧化应激;作用机制
KEYWORDS: curcumol;breast cancer cells T 47D;cell cycle arrest ;cell proliferation ;oxidative stress ;mechanism
阅读数: 179 次
本月下载数: 6 次

* 注:未经本站明确许可,任何网站不得非法盗链资源下载连接及抄袭本站原创内容资源!在此感谢您的支持与合作!