1Department of Oncology, The Third People's Hospital of Chengdu, Chengdu, Sichuan, 610031, China; 2Department of Oncology, The Affiliated Hospital of Southwest Jiaotong University, Chengdu, Sichuan, 610031, China.
Department of Laboratory Medicine, Eastern Hospital, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, Chengdu, Sichuan, 610101, China.
Acta Biochim Pol. 2022 Feb 28;69(1):139-145. doi: 10.18388/abp.2020_5776.
Lung cancer, a most common clinically diagnosed malignancy grows rapidly and undergoes metastasis/diffusion to distant organs at a fast rate. In the present study gravacridondiol tetrazole (tetrazole) was synthesized and investigated for lung cancer growth inhibition potential in vitro. MTT assay and flow cytometry using propidium iodide were used to determine viability changes and DNA content distribution. Protein expression and apoptotic changes were detected by western blotting and Annexin-V/PI assays. Treatment with 12 μM tetrazole suppressed viabilities to 23% and 20% in A549 and NCI-H1819 cells, respectively. In tetrazole exposed cells, G1-phase cell count increased significantly compared to the control. Tetrazole-treatment of A549 and NCI-H1819 cells caused a prominent raise in LC3‑II and p-ERK1/2 expression at 72 h. The SQSTM1/p62 level, p-mTOR and p-p70S6K expression was lowered significantly in A549 and NCI-H1819 cells on exposure to tetrazole. Exposure to U1026 alleviated tetrazole mediated LC3II/I ratio increase in A549 and NCI-H1819 cells significantly (P<0.02) compared to tetrazole treated cells. Treatment with tetrazole and 3‑MA in combination led a significant (P<0.02) elevation in A549 and NCI-H1819 cell apoptotic count relative to tetrazole (12 μM) alone treated cells. Moreover, tetrazole and 3‑MA combination increased cleavage of caspase‑3 to a greater extent compared to tetrazole. In summary, tetrazole manifested anti-proliferative effect on lung cancer cells via autophagy over-activation and arrest of cell cycle. It deactivated ERK1/2 signalling and promoted mTOR signaling in A549 and NCI-H1819 cells to regulate cancer proliferation. Thus, tetrazole needs to be studied further as an anti-proliferative agent for treatment of lung cancer.
肺癌是临床上最常见的恶性肿瘤之一，其生长迅速，并以很快的速度向远处器官转移/扩散。在本研究中，合成了并研究了gravacridondiol 四唑（四唑）对体外肺癌生长抑制的潜力。使用噻唑蓝（MTT）法和碘化丙啶（PI）流式细胞术检测细胞活力变化和 DNA 含量分布。通过 Western blot 和 Annexin-V/PI 检测法检测蛋白表达和凋亡变化。用 12 μM 四唑处理 A549 和 NCI-H1819 细胞，其活力分别降至 23%和 20%。与对照组相比，在暴露于四唑的细胞中，G1 期细胞计数明显增加。在 A549 和 NCI-H1819 细胞中，四唑处理 72 小时后，LC3-II 和 p-ERK1/2 的表达明显升高。SQSTM1/p62 水平、p-mTOR 和 p-p70S6K 表达在 A549 和 NCI-H1819 细胞暴露于四唑后明显降低。与单独用四唑处理的细胞相比，用 U1026 处理可显著减轻四唑介导的 A549 和 NCI-H1819 细胞 LC3II/I 比值的增加（P<0.02）。与单独用四唑（12 μM）处理的细胞相比，用四唑和 3-MA 联合处理可显著（P<0.02）增加 A549 和 NCI-H1819 细胞的凋亡计数。此外，与四唑相比，四唑和 3-MA 联合处理更能促进 caspase-3 的裂解。总之，四唑通过过度激活自噬和细胞周期阻滞，对肺癌细胞表现出抗增殖作用。它使 ERK1/2 信号失活，并在 A549 和 NCI-H1819 细胞中促进 mTOR 信号，从而调节癌症增殖。因此，四唑需要进一步研究作为一种抗增殖剂，用于治疗肺癌。
