南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (2): 201-206.doi: 10.12122/j.issn.1673-4254.2022.02.05

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萘烯丙基三氟甲基苯并环戊酮抑制肺癌A549细胞增殖并诱导其凋亡

吴 旭，何俊杰，李 萍，王 毅，颜 亮，马金珠   

  1. 皖南医学院活性生物大分子安徽省重点实验室，安徽 芜湖 241002；南京大学化学化工学院，江苏 南京 210000
  • 出版日期:2022-02-20 发布日期:2022-03-16

Naphthalene allyl trifluoromethyl benzocyclopentanone inhibits proliferation and induces apoptosis of lung cancer A549 cells in vitro

WU Xu, HE Junjie, LI Ping, WANG Yi, YAN Liang, MA Jinzhu   

  1. Anhui Key Laboratory of Active Biomacromolecules, Wannan Medical College, Wuhu 241002, China; School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210000, China
  • Online:2022-02-20 Published:2022-03-16

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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、25 μg/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（200 μL），对实验组注射化合物XX0335（12.5 μg/mL，200 μL），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。

关键词: 萘烯丙基三氟甲基苯并环戊酮；细胞增殖；细胞凋亡；Akt/mTOR信号通路

Abstract: 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 25 μg/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.

Key words: naphthalene allyl trifluoromethyl benzocyclopentanone; cell proliferation; cell cycle; apoptosis; Akt/mTOR signal pathway