螺吖啶化合物AMTAC - 06对HCT - 116结肠癌细胞的凋亡和抗氧化作用
Apoptotic and antioxidant effects in HCT-116 colorectal carcinoma cells by a spiro-acridine compound, AMTAC-06.
作者信息
Duarte Sâmia Sousa, Silva Daiana Karla Frade, Lisboa Thaís Mangeon Honorato, Gouveia Rawny Galdino, de Andrade Camyla Caroliny Neves, de Sousa Valgrícia Matias, Ferreira Rafael Carlos, de Moura Ricardo Olimpio, Gomes Joilly Nilce Santana, da Silva Patricia Mirella, de Lourdes Assunção Araújo de Azevedo Fátima, Keesen Tatjana S L, Gonçalves Juan Carlos Ramos, Batista Leônia Maria, Sobral Marianna Vieira
机构信息
Postgraduation Program in Bioactive Natural and Synthetic Products, Federal University of Paraíba, João Pessoa, Paraíba, Brazil.
Drug Development and Synthesis Laboratory, Department of Pharmacy, State University of Paraíba, João Pessoa, Paraíba, Brazil.
出版信息
Pharmacol Rep. 2022 Jun;74(3):545-554. doi: 10.1007/s43440-022-00357-0. Epub 2022 Mar 17.
BACKGROUND
Acridine compounds have been described as promising anticancer agents. Previous studies showed that (E)-1'-((4-chlorobenzylidene)amino)-5'-oxo-1',5'-dihydro-10H-spiro[acridine-9,2'-pyrrole]-4'-carbonitrile (AMTAC-06), a spiro-acridine compound, has antitumor activity on Ehrlich tumor and low toxicity. Herein, we investigated its antitumor effect against human cells in vitro.
METHODS
MTT assay was used to assess cytotoxicity of AMTAC-06 (3.125-200 µM) against tumor and non-tumor cells, and the half-maximal inhibitory concentration (IC) and the selectivity index (SI) were calculated. The effects on the cell cycle (propidium iodide-PI-staining), apoptosis (Annexin V-FITC/PI double staining by flow cytometry), and production of reactive oxygen species, ROS (DCFH assay) were also evaluated. Statistical analysis was achieved using ANOVA followed by Tukey's post-test.
RESULTS
AMTAC-06 showed higher cytotoxicity against colorectal carcinoma HCT-116 cells (IC: 12.62 µM). The SI showed that AMTAC-06 was more selective for HCT-116 cells (HaCaT SI: 1.41; PBMC SI: 0.62) than doxorubicin (HaCaT SI: 0.10; PBMC SI: 0.01). AMTAC-06 (15 and 30 µM) induced an increase in the sub-G1 peak (p < 0.000001) and cell cycle arrest in S phase (p = 0.003547). Moreover, treatment with this compound (15 and 30 µM) resulted in increased early (p < 0.000001) and late apoptotic cells (p < 0.000001). In addition, there was a reduction on ROS production (p < 0.000001).
CONCLUSIONS
AMTAC-06 presents anticancer activity against HCT-116 cells by regulating the cell cycle, inducing apoptosis and an antioxidant action.
背景
吖啶化合物已被描述为有前景的抗癌药物。先前的研究表明，一种螺吖啶化合物（E）-1'-（（4-氯亚苄基）氨基）-5'-氧代-1'，5'-二氢-10H-螺[吖啶-9，2'-吡咯]-4'-甲腈（AMTAC-06）对艾氏腹水瘤具有抗肿瘤活性且毒性较低。在此，我们研究了其对人细胞的体外抗肿瘤作用。
方法
采用MTT法评估AMTAC-06（3.125 - 200 μM）对肿瘤细胞和非肿瘤细胞的细胞毒性，并计算半数抑制浓度（IC）和选择性指数（SI）。还评估了其对细胞周期（碘化丙啶 - PI染色）、凋亡（通过流式细胞术进行膜联蛋白V - FITC/PI双染色）以及活性氧（ROS）产生（DCFH检测）的影响。使用方差分析（ANOVA）及随后的Tukey事后检验进行统计分析。
结果
AMTAC-06对结肠直肠癌HCT-116细胞显示出更高的细胞毒性（IC：12.62 μM）。选择性指数表明，AMTAC-06对HCT-116细胞的选择性高于阿霉素（HaCaT细胞的SI：1.41；PBMC细胞的SI：0.62，而阿霉素HaCaT细胞的SI：0.10；PBMC细胞的SI：0.01）。AMTAC-06（15和30 μM）诱导亚G1峰增加（p < 0.000001）并使细胞周期停滞于S期（p = 0.003547）。此外，用该化合物（15和30 μM）处理导致早期凋亡细胞增加（p < 0.000001）和晚期凋亡细胞增加（p < 0.000001）。另外，ROS产生减少（p < 0.000001）。
结论
AMTAC-06通过调节细胞周期、诱导凋亡和抗氧化作用对HCT-116细胞呈现抗癌活性。
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