摘要
目的研究一种新型二氢吡喃并[2,3-c]吡唑类衍生物(36号化合物)的体外抑瘤作用及可能的分子机制。方法 MTT比色法检测36号化合物对人乳腺癌细胞Bcap-37,MCF-7和人肺癌细胞A549体外存活的影响,反转录荧光定量PCR法检测Bcap-37细胞凋亡相关基因p53,p21,Bax和NF-κB mRNA水平,流式细胞术检测Bcap-37细胞周期和细胞凋亡,JC-1荧光探针法测定Bcap-37细胞线粒体膜电位(MMP)。结果 36号化合物对Bcap-37,MCF-7和A549细胞存活均表现出浓度依赖性抑制作用,其中对人乳腺癌细胞Bcap-37的半数抑制浓度(IC50)最低,为(22.4±0.8)mg·L^(-1)。与正常对照组相比,36号化合物在25 mg·L^(-1)浓度下使Bcap-37细胞周期阻滞于G1期(P<0.01),早期凋亡细胞显著增加(P<0.01);在50 mg·L^(-1)浓度下,使Bcap-37细胞凋亡相关基因p53,p21,Bax和NF-κB mRNA水平均显著升高(P<0.05,P<0.01);在10,25和75 mg·L^(-1)浓度下,使Bcap-37细胞MMP明显降低(P<0.05,P<0.01)。结论 36号化合物具有良好的体外抑瘤作用,可抑制Bcap-37细胞增殖,促进其凋亡,其作用机制与细胞内线粒体通路和细胞外凋亡信号通路的激活相关。
OBJECTIVE To investigate the anti-tumor effect in vitro and underlying mechanism of a novel dihydropyran [2,3-c] pyrazole derivative(No.36 Compound).METHODS MTT assay was used to determine the proliferation activity on Bcap-37,MCF-7 and A549 cancer cell lines.The mRNA levels of apoptosis-related genes including p53,p21,Bax and NF-κB on Bcap-37 cells were detected by RT-PCR.The cell cycle and apoptosis of Bcap-37 cells were determined by flow cytometry.The mitochondrial membrane potential(MMP) of Bcap-37 cells was measured by JC-1 fluorescent probe.RESULTS No.36 Compound exhibited inhibition on the in vitro proliferation of Bcap-37,MCF-7 and A549 cells.The half maximal inhibitory concentration(IC50) of No.36 Compound acting on Bcap-37 cells for 48 h was(22.4±0.8)mg·L^-1.Compared with the normal control group,No.36 Compound 25 mg·L^-1 induced G1 arrest in Bcap-37 cells(P<0.01).Consistently,early apoptotic cells were increased markedly(P<0.01).Furthermore,the relative expressions of apoptosis-related genes p53,p21,Bax and NF-κB in Bcap-37 cells were elevated(P<0.05,P<0.01) after treatment with No.36 Compound 50 mg·L^-1.The MMP of Bcap-37 cel s was significantly reduced(P<0.05,P<0.01) after being treated with No.36 Compound10,25 and 75 mg·L^-1.CONCLUSION No.36 Compound has potential in vitro anti-tumor activity,which may be related to the activation of the intracellular mitochondrial pathway and extracellular apoptotic signal pathway.
作者
高梦诗
郑敏子
叶心怡
施国邦
阙祥洁
张隆超
孙漩嵘
GAO Meng-shi;ZHENG Min-zi;YE Xin-yi;SHI Guo-bang;QUE Xiang-jie;ZHANG Long-chao;SUN Xuan-rong(Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals,Zhejiang University of Technology,Hangzhou 310014,China)
出处
《中国药理学与毒理学杂志》
CAS
CSCD
北大核心
2018年第8期614-620,共7页
Chinese Journal of Pharmacology and Toxicology
基金
国家自然科学基金(21506192)
浙江省自然科学基金(LY16E030010)~~
