Department of Pediatric Hematology/Oncology, West China Second University Hospital, Sichuan University, Chengdu 610041, Sichuan, China.
Biomed Environ Sci. 2013 Nov;26(11):902-11. doi: 10.3967/bes2013.019.
To explore the effects of resveratrol-induced apoptosis and autophagy in T-cell acute lymphoblastic leukemia (T-ALL) cells and potential molecular mechanisms.
The anti-proliferation effect of resveratrol-induced, apoptosis and autophagy on T-ALL cells were detected by using MTT test, immunofluorescence, electronic microscope, and flow cytometry, respectively. Western blotting was performed for detecting changes of apoptosis-associated proteins, cell cycle regulatory proteins and state of activation of Akt, mTOR, p70S6K, 4E-BP1, and p38-MAPK.
Resveratrol inhibited the proliferation and induced apoptosis and autophagy in T-ALL cells in a dose and time-dependent manner. It also induced cell cycle arrest at G0/G1 phase via up regulating cyclin-dependent kinase (CDK) inhibitors p21 and p27 and down regulating cyclin A and cyclin D1. Western blotting revealed that resveratrol significantly decreased the expression of antiapoptotic proteins (Mcl-1 and Bcl-2) and increased the expression of proapoptotic proteins (Bax, Bim, and Bad), and induced cleaved-caspase-3 in a time-dependent manner. Significant increase in ratio of LC3-II/LC3-I and Beclin 1 was also detected. Furthermore, resveratrol induced significant dephosphorylation of Akt, mTOR, p70S6K, and 4E-BP1, but enhanced specific phosphorylation of p38-MAPK which could be blocked by SB203580. When autophagy was suppressed by 3-MA, apoptosis in T-ALL cells induced by resveratrol was enhanced.
Our findings have suggested that resveratrol induces cell cycle arrest, apoptosis, and autophagy in T-ALL cells through inhibiting Akt/mTOR/p70S6K/4E-BP1 and activating p38-MAPK signaling pathways. Autophagy might play a role as a self-defense mechanism in T-ALL cells treated by resveratrol. Therefore, the reasonable inhibition of autophagy in T-ALL cells may serve as a promising strategy for resveratrol induced apoptosis and can be used as adjuvant chemotherapy for T-ALL.
探讨白藜芦醇诱导 T 细胞急性淋巴细胞白血病（T-ALL）细胞凋亡和自噬的作用及其潜在的分子机制。
采用 MTT 试验、免疫荧光、电子显微镜和流式细胞术分别检测白藜芦醇诱导的 T-ALL 细胞增殖抑制、凋亡和自噬作用，Western blot 检测凋亡相关蛋白、细胞周期调控蛋白及 Akt、mTOR、p70S6K、4E-BP1、p38-MAPK 活化状态的变化。
白藜芦醇呈剂量和时间依赖性抑制 T-ALL 细胞增殖，并诱导其凋亡和自噬，同时通过上调细胞周期蛋白依赖性激酶（CDK）抑制剂 p21 和 p27、下调细胞周期蛋白 A 和 D1 诱导细胞周期 G0/G1 期阻滞。Western blot 显示，白藜芦醇显著降低抗凋亡蛋白（Mcl-1 和 Bcl-2）的表达，增加促凋亡蛋白（Bax、Bim 和 Bad）的表达，并呈时间依赖性诱导 cleaved-caspase-3 的表达。还检测到 LC3-II/LC3-I 比值和 Beclin 1 的显著增加。此外，白藜芦醇诱导 Akt、mTOR、p70S6K 和 4E-BP1 的显著去磷酸化，但增强了 p38-MAPK 的特异性磷酸化，该作用可被 SB203580 阻断。当自噬被 3-MA 抑制时，白藜芦醇诱导的 T-ALL 细胞凋亡增强。
本研究结果提示，白藜芦醇通过抑制 Akt/mTOR/p70S6K/4E-BP1 并激活 p38-MAPK 信号通路诱导 T-ALL 细胞发生细胞周期阻滞、凋亡和自噬。自噬可能在白藜芦醇处理的 T-ALL 细胞中作为一种自我保护机制发挥作用。因此，在 T-ALL 细胞中合理抑制自噬可能成为白藜芦醇诱导凋亡的一种有前途的策略，并可作为 T-ALL 的辅助化疗。
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