摘要
目的探讨新型萘烯丙基三氟甲基苯并环戊酮(XX0335)抑制肺癌细胞系A549增殖并诱导凋亡的分子机制。方法实验分为4组:对照组(含0.1%DMSO的培养液)、另外3组为不同浓度(6.25、12.5、25μg/mL)的XX0335(该化合物需DMSO溶解,但工作浓度中DMSO的终浓度均低于0.1%)。通过CCK-8和EdU实验分别测定各组对A549细胞活力及增殖的影响,通过流式细胞术测定上述浓度的XX0335对A549细胞凋亡及周期的影响;通过免疫印迹实验测定不同浓度的XX0335对增殖相关蛋白Akt、mTOR、Akt/mTOR磷酸化水平、凋亡相关蛋白cleaved PARP及细胞周期相关蛋白CyclinD1表达水平的影响;通过裸鼠荷瘤实验探究XX0335对A549增殖的影响,动物实验使用4周龄小鼠共10只,随机均分为2组。将2×106A549细胞植入小鼠左侧腋下,待26 d后瘤体长径长至1 cm时,对照组小鼠注射无菌生理盐水(200μL),对实验组注射化合物XX0335(12.5μg/mL,200μL),隔天注射1次,共5次之后取肿瘤观察每组瘤体大小。结果CCK-8实验表明,A549细胞活力在XX0335的作用下比对照组呈剂量依赖性下降(P<0.01)。EdU实验发现XX0335能显著抑制A549细胞增殖(P<0.001)。流式细胞术检测结果表明,细胞凋亡较对照组呈剂量依赖性上升(P<0.01),且细胞停滞于G1期。与对照组相比,AKT、mTOR的磷酸化受到了明显抑制,cleaved PARP表达量升高,且CyclinD1的蛋白表达量下降。裸鼠荷瘤实验结果显示注射XX0335后肿瘤体积明显减小(P<0.01)。结论XX0335抑制了肺癌A549细胞增殖并诱导细胞发生了凋亡,且细胞周期阻滞于G1期,其作用机制可能与抑制Akt/mTOR信号通路有关。
Objective To investigate the molecular mechanism by which a novel naphthalene allyl trifluoromethyl benzocyclopentanone XX0335 inhibits the proliferation and induces apoptosis of lung cancer A549 cells.Methods Lung cancer A549 cells were treated with 0.1%DMSO(control)or different concentrations(6.25,12.5,and 25μg/mL)of XX0335,and the changes in cell viability,cell cycle,proliferation and apoptosis were assessed with CCK-8 assay,EdU experiment,and flow cytometry.The effects of different concentrations of XX0335 on phosphorylation levels of proliferation-related proteins Akt,mTOR,Akt/mTOR and the expressions of cleaved PARP and cyclin D1 were determined using Western blotting.We also assessed the effect of XX0335 on tumor growth in a mouse model bearing A945 cell xenograft.Results Treatment with XX0335 reduced the viability of A549 cells in a dose-dependent manner(P<0.01)and significantly inhibited cell proliferation(P<0.001).Flow cytometry showed that XX0335 treatment promoted apoptosis of the cells(P<0.01)and caused an obvious increase of the number of G1-phase cells.Compared with DMSO,XX0335 significantly inhibited the phosphorylation of Akt and mTOR,increased the expression of cleaved PARP,and lowered the protein expression of cyclin D1.In the tumor-bearing mouse models,injection of XX0335 significantly decreased the tumor volume(P<0.01).Conclusion XX0335 inhibits the proliferation,cycle and induces apoptosis of lung cancer A549 cells possibly by inhibiting the Akt/mTOR signal pathway.
作者
吴旭
何俊杰
李萍
王毅
颜亮
马金珠
WU Xu;HE Junjie;LI Ping;WANG Yi;YAN Liang;MA Jinzhu(Anhui Key Laboratory of Active Biomacromolecules,Wannan Medical College,Wuhu 241002,China;School of Chemistry and Chemical Engineering,Nanjing University,Nanjing 210000,China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2022年第2期201-206,共6页
Journal of Southern Medical University
基金
国家自然科学基金(22071101)
安徽省教育厅自然科学重点项目(KJ2021A0852、KJ2021A0856)。
