Piperlongumine Induces Apoptosis and Pyroptosis of NCI-H460 Lung Cancer Cells Via Increasing ROS
JIN Lei, LUO Zhen-yu, LÜ Ang
Department of Pharmacy, Key Laboratory of Clinical Cancer Pharmacology and Toxicology Research of Zhejiang Province, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou 310006, China
Abstract：OBJECTIVE To investigate the inhibitory effect of different concentration of piperlongumine on the growth of lung cancer cells, and further study the effect of piperlongumine on the apoptosis and pyroptosis of NCI-H460 cancer cells by increasing ROS, and elucidate its new anti-tumor mechanism. METHODS NCI-H460 lung cancer cells were cultured by conventional method, the control group and different concentration of piperlongumine treatment group were set up. MTT assay was used to evaluate the activity of NCI-H460 lung cancer cells, and cell colony assay was performed for the effect of colony formation of NCI-H460 cells. The NCI-H460 cells apoptosis and the growth inhibition of NCI-H460 cells via promotion of ROS levels were evaluated by flow cytometry, respectively. Western blotting was used to detect the expression of Bax, Bcl-2 and caspase-3 proteins after treated with piperlongumine. The effect of piperlongumine on the ROS-based NCI-H460 lung cells pyroptosis was confirmed by microscopic imaging. At the same time, the effect of ROS on apoptosis-related protein GSDME was detected by Western blotting. RESULTS Piperlongumine inhibited lung cancer cells in a concentration dependent manner, with the IC50 value on NCI-H460 lung cancer of 13.1 μmol.L-1. Piperlongumine restrained the NCI-H460 colony formation in a concentration-dependent pattern and promoted NCI-H460 apoptosis. Piperlongumine promoted the expression of the pro-apoptotic proteins Bax and Cle-caspase-3, and inhibited the expression of the anti-apoptotic protein Bcl-2 in a concentration-dependent manner. Piperlongumine induced NCI-H460 cells apoptosis and pyroptosis by increasing intracellular ROS content, and ROS increased GSDME-N protein expression. CONCLUSION Piperlongumine inhibits the growth of lung cancer cells, and induces apoptosis and pyroptosis of NCI-H460 cancer cells by promoting the levels of intracellular ROS.
金磊, 罗震宇, 吕昂. 荜拔酰胺对活性氧诱导肺癌NCI-H460细胞凋亡与焦亡的影响[J]. 中国药学杂志, 2020, 55(12): 1002-1007.
JIN Lei, LUO Zhen-yu, LÜ Ang. Piperlongumine Induces Apoptosis and Pyroptosis of NCI-H460 Lung Cancer Cells Via Increasing ROS. Chinese Pharmaceutical Journal, 2020, 55(12): 1002-1007.
DHOLARIA B, HAMMOND W, SHREDERS A, et al. Emerging therapeutic agents for lung cancer[J]. J Hematol Oncol, 2016, 9(1):138-151.
[2]
HIRSCH F R, SCAGLIOTTI G V, MULSHINE J L, et al. Lung cancer: current therapies and new targeted treatments[J]. Lancet, 2017, 389(10066):299-311.
[3]
YAN D D, ZHOU J S, XU X X, et al. Efficacy and toxicity of three different concurrent chemoradiotherapy in local advanced cervical cancer[J]. Chin J Mod Appl Pharm(中国现代应用药学), 2017, 34(9):1317-1321.
[4]
LIU J, LIU W, LU Y, et al. Piperlongumine restores the balance of autophagy and apoptosis by increasing Bcl-2 phosphorylation in rotenone-induced Parkinson disease models[J]. Autophagy, 2018, 14(5):845-861.
[5]
WU Y, MIN X, ZHUANG C, et al. Design, synthesis and biological activity of piperlongumine derivatives as selective anticancer agents[J]. Eur J Med Chem, 2014, 82:545-551.
[6]
XU S, WANG C. Piperlongumine induces apoptosis of lung cancer cells via the PI3K/Akt/mTOR signaling pathways[J]. J Qingdao Univ Sci Tech(Nat Sci Ed) (青岛科技大学学报:自然科学版), 2018, 39(3):51-56.
[7]
ZHENG J, SON D J, GU S M, et al. Piperlongumine inhibits lung tumor growth via inhibition of nuclear factor kappa B signaling pathway[J]. Sci Rep, 2016, 6:26357.
[8]
HONG Y, SENGUPTA S, HUR W, et al. Identification of novel ROS inducers: quinone derivatives tethered to long hydrocarbon chains[J]. J Med Chem, 2015, 58(9):3739-3750.
[9]
ZHENG J, SON D J, GU S M, et al. Piperlongumine inhibits lung tumor growth via inhibition of nuclear factor kappa B signaling pathway[J]. Sci Rep, 2016, 6:26357.
[10]
THONGSOM S, SUGINTA W, LEE K J, et al. Piperlongumine induces G2/M phase arrest and apoptosis in cholangiocarcinoma cells through the ROS-JNK-ERK signaling pathway[J]. Apoptosis, 2017, 22(11):1473-1484.
[11]
JIN H O, LEE Y H, PARK J A, et al. Piperlongumine induces cell death through ROS-mediated CHOP activation and potentiates TRAIL-induced cell death in breast cancer cells[J]. J Cancer Res Clin Oncol, 2014, 140(12):2039-2046.
[12]
PHILP N H, DILLONC P, SNYDER A G, et al. Caspase-8 mediates caspase-1 processing and innate immune defense in response to bacterial blockade of NF-κB and MAPK signaling[J]. Proc Natl Acad Sci USA, 2014, 111(20):7385-7390.
[13]
WANGY, GAO W, SHI X, et al. Chemotherapy drugs induce pyroptosis through caspase-3 cleavage of a gasdermin[J]. Nature, 2017, 547(7661):99-103.
[14]
WANG W J, CHEN D, JIANG M Z, et al. Downregulation of gasdermin D promotes gastric cancer proliferation by regulating cell cycle-related proteins[J]. J Dig Dis, 2018, 19(2):74-83.
[15]
SHARMA D, KANNEGANTI T D. Inflammatory cell death in intestinal pathologies[J]. Immunol Rev, 2017, 280(1):57-73.
[16]
MAN S M. Inflammasomes in the gastrointestinal tract: infection, cancer and gut microbiota homeostasis[J]. Nat Rev Gastroenterol Hepatol, 2018, 15(12):721-737.
[17]
ZHAI Z, LIU W, KAUR M, et al. NLRP1 promotes tumor growth by enhancing inflammasome activation and suppressing apoptosis in metastatic melanoma[J]. Oncogene, 2017, 36(27):3820-3830.
[18]
CHOI J B, KIM J H, LEE H, et al. Reactive oxygen species and p53 mediated activation of p38 and caspases is critically involved in kaempferol induced apoptosis in colorectal cancer cells[J]. J Agric Food Chem, 2018, 66(38):9960-9967.
2020, Vol. 55 
