2,3-吲哚醌对乳腺癌BT549细胞生长的抑制作用及其机制  

INHIBITORY EFFECT OF 2,3-INDOLEQUINONE ON THE GROWTH OF BREAST CANCER BT549 CELLS AND ITS MECHANISM

作　　者：李雪 杨雨 张丽[3] 周娜 侯琳 LI Xue;YANG Yu;ZHANG Li;ZHOU Na;HOU Lin(School of Basic Medicine, Qingdao University, Qingdao 266021, China)

机构地区：[1]青岛大学医学部基础医学院,山东青岛266021 [2]中山大学肿瘤防治中心神经外科 [3]青岛大学药学院

出　　处：《精准医学杂志》2020年第3期208-212,共5页Journal of Precision Medicine

基　　金：国家自然科学基金资助项目(81472542);山东省重点研发项目(2019GSF107025);青岛大学医学院创新团队青年教师培育项目(600201304)。

摘　　要：目的探讨2,3-吲哚醌对乳腺癌BT549细胞生长的抑制作用及其机制。方法体外培养乳腺癌BT549细胞,通过CCK8实验筛选2,3-吲哚醌药物浓度并检测其对BT549细胞增殖的抑制作用;乳腺癌BT549细胞经2,3-吲哚醌加药处理72 h后,通过Transwell、划痕实验检测2,3-吲哚醌对BT549细胞侵袭和迁移的抑制作用;2,3-吲哚醌加药处理BT549细胞72 h后,采用实时定量PCR(qPCR)方法检测BT549细胞中p53基因(p53)、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白基因(Bax)以及MDM2基因(MDM2)mRNA相对表达量的变化;2,3-吲哚醌加药处理BT549细胞72 h后,采用Western blot方法检测细胞中P53、Bcl-2、Bax和MDM2蛋白的相对表达情况。结果CCK8实验结果显示,与0μmol/L组相比,随着2,3-吲哚醌浓度的升高,BT549细胞的增殖水平逐渐下降(F=74.64,P<0.01)。Transwell实验和划痕实验结果显示,与0μmol/L组相比,加入2,3-吲哚醌的浓度越高,细胞的侵袭和迁移能力越低(F=751.48、314.67,P<0.01)。qPCR方法检测结果显示,与0μmol/L组相比较,随着2,3-吲哚醌浓度的增加,细胞中p53和Bax mRNA相对表达量逐渐增高(F=62.82、20.86,P<0.01),Bcl-2、MDM2 mRNA相对表达量逐渐下降(F=21.53、30.31,P<0.01)。Western blot检测结果显示,与0μmol/L组比较,随着2,3-吲哚醌浓度的增高,细胞中P53、Bax蛋白的相对表达水平逐渐增高(F=29.59、53.12,P<0.01),Bcl-2、MDM2蛋白相对表达水平逐渐下降(F=14.85、159.81,P<0.01)。结论2,3-吲哚醌通过P53信号通路抑制乳腺癌BT549细胞的增殖、侵袭和迁移。Objective To investigate the inhibitory effect of 2,3-indolequinone on the growth of breast cancer BT549 cells and its mechanism.Methods Breast cancer BT549 cells were cultured in vitro.CCK8 assay was used to screen the concentrations of 2,3-indolequinone and evaluate its inhibitory effect on the proliferation of BT549 cells.After the BT549 cells were treated with 2,3-indolequinone for 72 h,the inhibitory effects of 2,3-indolequinone on the invasion and migration of BT549 cells were examined by Transwell and wound healing assays,respectively.Also,after the BT549 cells were treated with 2,3-indolequinone for 72 h,the relative mRNA and protein expression levels of p53,Bcl-2,Bax,and MDM2 in BT549 cells were measured by real-time quantitative PCR(qPCR)and Western blot,respectively.Results The proliferation of BT549 cells gradually decreased with an increase in 2,3-indolequinone concentration(F=74.64,P<0.01),according to the CCK8 assay.The Transwell and wound healing assays showed that the higher the concentration of 2,3-indolequinone added,the lower the invasion and migration ability of cells(F=751.48,314.67,P<0.01).The results of qPCR showed that with an increase in 2,3-indolequinone concentration,the relative expression of p53 and Bax mRNA in the cells gradually increased(F=62.82,20.86,P<0.01),while the relative expression of Bcl-2 and MDM2 mRNA gradually decreased(F=21.53,30.31,P<0.01).The results of Western blot showed that with an increase in 2,3-indolequinone concentration,the relative expression levels of P53 and Bax proteins in cells gradually increased(F=29.59,53.12,P<0.01),while the relative expression levels of Bcl-2 and MDM2 proteins gradually decreased(F=14.85,159.81,P<0.01).Conclusion 2,3-Indolequinone inhibits the proliferation,invasion,and migration of breast cancer BT549 cells through the P53 signaling pathway.

关 键 词：乳腺肿瘤 2 3-吲哚醌 基因 p53 肿瘤抑制蛋白质P53 bcl-2 原癌基因蛋白质C-BCL-2 BCL-2相关X蛋白质 细胞增殖 细胞运动 体外研究 

分 类 号：R737.9[医药卫生—肿瘤] R329.2[医药卫生—临床医学]