Lu Li, Yang Li-Ning, Wang Xue-Xi, Song Chun-Li, Qin Hong, Wu Yong-Jie
Department of Pharmacology, School of Medicine, Lanzhou University, Key Laboratory of Preclinical Study for New Drugs of Gansu Province, Lanzhou, 730000, China.
Department of Pharmacy, The Second Hospital of Gansu Province, Lanzhou, 730000, China.
Chin J Integr Med. 2017 Feb;23(2):125-131. doi: 10.1007/s11655-016-2591-1. Epub 2016 Jun 14.
To evaluate the cytotoxic effects of ampelopsin sodium (Amp-Na) and carboplatin (CBP) used alone or in combination on human non-small cell lung cancer (NSCLC) cells SPC-A1 in vitro and its related mechanism.
Cytotoxic effects were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. The synergistic effects of the drugs were calculated with coefficient of drug interaction (CDI). Cell cycle was determined by flow cytometry (FCM). The levels of p53, p21, cyclinE, cyclinD1, and phosphorylated cyclin-dependent kinase-2 (p-CDK2) were evaluated by Western blot.
Amp-Na (6.25-200 μg/mL) and CBP (3.13-100 μg/mL) alone exhibited prominent cytotoxic activity in a concentration-dependent manner on SPC-A1 cells with 50% inhibitive concentration values of 57.07±14.46 and 34.97±6.30 μg/mL, respectively. Drug combinations were associated with significantly higher cytotoxic effects than each drug alone (P<0.05 or 0.01). The CDI analysis confirmed the synergy of Amp-Na and CBP on inhibiting cancer cell viability across a wide concentration range (CDI <1). FCM and Western blot showed that synergistic cytotoxic effects of Amp-Na and CBP were related to G arrested which mainlym ediated by p 21 through the inhibition of CDK2 activity independent of the p53 tumor suppressor pathway.
Amp-Na exhibits anticancer activities and enhances the antitumor activities of CBP through up-regulation of p21 and inhibition of CDK2 activity in human NSCLC cells SPC-A1. These results suggest that Amp-Na may be applied to enhance the anticancer action of CBP.
评估蛇葡萄素钠（Amp-Na）与卡铂（CBP）单独及联合应用对人非小细胞肺癌（NSCLC）细胞SPC-A1的体外细胞毒性作用及其相关机制。
采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）法评估细胞毒性作用。用药物相互作用系数（CDI）计算药物的协同作用。通过流式细胞术（FCM）测定细胞周期。采用蛋白质免疫印迹法检测p53、p21、细胞周期蛋白E、细胞周期蛋白D1及磷酸化细胞周期蛋白依赖性激酶-2（p-CDK2）的水平。
单独使用Amp-Na（6.25 - 200 μg/mL）和CBP（3.13 - 100 μg/mL）对SPC-A1细胞均表现出显著的浓度依赖性细胞毒性活性，其半数抑制浓度值分别为57.07±14.46和34.97±6.30 μg/mL。联合用药的细胞毒性作用明显高于单药（P<0.05或0.01）。CDI分析证实Amp-Na和CBP在较宽浓度范围内对抑制癌细胞活力具有协同作用（CDI<1）。FCM和蛋白质免疫印迹法显示，Amp-Na和CBP的协同细胞毒性作用与G期阻滞有关，主要通过p21介导，抑制CDK2活性，且不依赖于p53肿瘤抑制途径。
Amp-Na具有抗癌活性，可通过上调p21和抑制CDK2活性增强CBP对人NSCLC细胞SPC-A1的抗肿瘤活性。这些结果表明Amp-Na可用于增强CBP的抗癌作用。
