Liu Ying, Jiang Zi-Yu, Zhou Yuan-Li, Qiu Hui-Hui, Wang Gang, Luo Yi, Liu Jing-Bing, Liu Xiong-Wei, Bu Wei-Quan, Song Jie, Cui Li, Jia Xiao-Bin, Feng Liang
Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu Nanjing, 210028, PR China; School of Pharmacy, Anhui University of Chinese Medicine, Anhui Hefei 230038, PR China.
Key Laboratory of New Drug Delivery Systems of Chinese Materia Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu Nanjing, 210028, PR China; Third School of Clinical Medical of Nanjing University of Chinese Medicine, Jiangsu Nanjing 210028, PR China.
Biomed Pharmacother. 2017 Sep;93:490-497. doi: 10.1016/j.biopha.2017.06.073. Epub 2017 Jun 30.
Endoplasmic reticulum stress (ERs) has been regarded as an important cause for the pathogenesis of non-small-cell lung cancer (NSCLC). β-elemene is an active component in the essential oil extracted from a medicinal herb, Curcuma wenyujin, and has been reported to be effective against non-small-cell lung cancer (NSCLC). However, the potential effect and underlying mechanisms of β-elemene on regulating ERs to inhibit NSCLC are still unclear. In the present study, A549 cells and Lewis tumor-bearing C57BL/6J mice were established to evaluate this effect. Visualsonics Vevo 2100 Small Animal Dedicated High-frequency Color Ultrasound was performed to observe tumor volume in vivo. 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) was used to evaluate cell vitality of A549 cells. Furthermore, western blotting (WB), immunohistochemistry (IHC) and quantitative reverse transcription polymerase chain reaction (q-PCR) were applied to detect the ERs-related proteins. Flow cytometry was also applied to detect cell apoptosis and assay kit for reactive oxygen species (ROS) generation. Our results showed that β-elemene inhibited lung cancer tumor growth and cell vitality in a dose- and time-dependent manner. Not only that, β-elemene could up-regulate ERs-related proteins like PERK, IRE1α, ATF6, ATF4, CHOP and down-regulate the Bcl-2 expression. More importantly, ERs inhibitor 4-PBA, IRE1α inhibitor STF-083010, ATF6 inhibitor Anti-ATF6 and PERK inhibitor GSK2656157 can all reduce the amplitude of protein expression changes and apoptosis rates, then weaken the anti-tumor effect of β-elemene. Therefore, the present in vivo and in vitro study revealed that the anti-NSCLC effect of β-elemene is closely related to the activation of ERs through PERK/IRE1α/ATF6 pathway, and this might be beneficial for clinical therapy of NSCLC.
内质网应激（ERs）被认为是非小细胞肺癌（NSCLC）发病机制的一个重要原因。β-榄香烯是从药用植物温郁金中提取的挥发油中的一种活性成分，据报道对非小细胞肺癌（NSCLC）有效。然而，β-榄香烯调节内质网应激以抑制非小细胞肺癌的潜在作用和潜在机制仍不清楚。在本研究中，建立了A549细胞和荷Lewis瘤的C57BL/6J小鼠模型来评估这种作用。使用Visualsonics Vevo 2100小动物专用高频彩色超声观察体内肿瘤体积。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A549细胞的活力。此外，采用蛋白质免疫印迹法（WB）、免疫组织化学法（IHC）和定量逆转录聚合酶链反应（q-PCR）检测内质网应激相关蛋白。还采用流式细胞术检测细胞凋亡和活性氧（ROS）生成检测试剂盒。我们的结果表明，β-榄香烯以剂量和时间依赖性方式抑制肺癌肿瘤生长和细胞活力。不仅如此，β-榄香烯可上调PERK、IRE1α、ATF6、ATF4、CHOP等内质网应激相关蛋白，并下调Bcl-2表达。更重要的是，内质网应激抑制剂4-PBA、IRE1α抑制剂STF-083010、ATF6抑制剂Anti-ATF6和PERK抑制剂GSK2656157均可降低蛋白表达变化幅度和凋亡率，进而削弱β-榄香烯的抗肿瘤作用。因此，本体内外研究表明，β-榄香烯的抗非小细胞肺癌作用与通过PERK/IRE1α/ATF6途径激活内质网应激密切相关，这可能对非小细胞肺癌的临床治疗有益。
