摘要
目的探讨马尼地平增强阿霉素抗皮肤鳞状细胞癌的作用及可能的机制。方法体外培养A431细胞,实验分为对照组、阿霉素(0.01μmol/L)组、阿霉素和马尼地平(2.5、5、10μmol/L)联合给药组;MTT实验检测细胞增殖活力;细胞克隆形成实验检测细胞的增殖能力;荧光显微镜观察阿霉素进入细胞核的荧光强度;流式细胞术检测细胞凋亡及细胞内阿霉素的含量;Western blot实验检测AKT、ERK、STAT5及磷酸化蛋白p-AKT、p-ERK、p-STAT5的表达情况。结果 MTT法结果显示,马尼地平(2.5、5、10μmol/L)与阿霉素联用均可增加A431细胞对阿霉素的敏感性;细胞克隆形成实验结果显示,马尼地平(2.5、5、10μmol/L)与阿霉素联用能抑制A431细胞克隆的形成,而单用马尼地平或阿霉素时A431细胞克隆的形成无明显变化;荧光显微镜观察结果显示,马尼地平(10μmol/L)能增加阿霉素在A431细胞内的分布;流式细胞术检测结果显示,马尼地平(2.5、5、10μmol/L)与阿霉素联用能促进A431细胞的凋亡;Western blot实验结果显示,马尼地平(2.5、5、10μmol/L)与阿霉素联用能降低A431细胞p-AKT、p-ERK、p-STAT5蛋白的表达水平;当AKT/ERK/STAT5信号通路的活性被0.1μmol/L博舒替尼抑制后,5μmol/L马尼地平不能发挥阿霉素对皮肤鳞状细胞癌的增敏作用。结论马尼地平可能通过调节A431细胞的AKT/ERK/STAT5信号通路从而增强阿霉素抗皮肤鳞状细胞癌的作用。
Objective To investigate the effects of manidipine(M) on enhancing doxorubicin(DOX) against cutaneous squamous cell carcinoma(CSCC) and its possible mechanisms.Methods A431 cells were cultured in vitro and divided into the following groups: control(Ctr),DOX(0.01 μmol/L),DOX and M(2.5,5,10 μmol/L) combined group.MTT assay was used to detect cell proliferation activity.The long-term combine effects were further evaluated using colony formation assays.The fluorescence intensity of DOX entering the nucleus was observed under fluorescence microscope.Cell apoptosis and intracellular DOX contents were detected by flow cytometry.The expression levels of AKT,ERK,STAT5 and phosphorylated proteins were detected by Western blotting assay.Results MTT assay showed that M combined with DOX could increase the sensitivity of A431 cells to DOX.The cell colony formation experiment showed that the combination of M and DOX could inhibit the colony formation of A431 cells, but there was no significant change when M or DOX alone was used.The results of fluorescence microscopy showed that M(10 μmol/L) could increase the DOX distribution in A431 cells.Flow cytometry showed that the combination of M and DOX could promote the apoptosis of A431 cells.Western blotting results showed that the combination of M and DOX could reduce the expression levels of p-AKT,p-ERK and p-STAT5 proteins in A431 cells;When the activity of AKT/ERK/STAT5 signaling pathway was inhibited by 0.1 μmol/L bosutinib, 5 μmol/L manidipine could not exert the sensitization effect of doxorubicin on A431 cells.Conclusion Manidipine may modulate the AKT/ERK/STAT5 signaling pathway in A431 cells to enhance the activity of doxorubicin against cutaneous squamous cell carcinoma.
作者
冯森玲
袁中文
林颖
晏烽根
曹礼慧
黄鸿亮
梁宝莹
FENG Senling;YUAN Zhongwen;LIN Ying;YAN fenggen;CAO Lihui;HUANG Hongliang;LIANG Baoying(Department of Pharmacy,Key Laboratory for Major Obstetric Diseases of Guangdong Province,the Third Affiliated Hospital of Guangzhou Medical University,Guangzhou 510150,China;Guangdong Provincial Clinical Research Center for Chinese Medicine Dermatology,Department of Dermatology,the Second Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangdong Provincial Hospital of Traditional Chinese Medicine,Guangzhou 510120,China)
出处
《中国皮肤性病学杂志》
CAS
CSCD
北大核心
2021年第11期1231-1237,共7页
The Chinese Journal of Dermatovenereology
基金
国家自然科学基金(82104446)
广东省基础与应用基础研究基金(2019A1515111109)
毅文人才工程
贺林院士新医学科研基金(2021HLKY02)
广东省中医皮肤临床医学研究中心及广东省中医皮肤病研究所专项(PFK2021-03)
广东省医学科学技术研究基金(A2020365)。
