白术内酯Ⅰ通过TLR4/MyD88通路调控肺癌A549细胞增殖侵袭的研究
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摘要
目的探究白术内酯Ⅰ在肺癌进程中的作用及分子机制。方法用qRT-PCR和免疫组化实验检测肺癌组织和癌旁组织中TLR4及MyD88的mRNA和蛋白的表达;Transwell侵袭实验及MTT实验检测白术内酯Ⅰ(100μM/L)对A549细胞侵袭、增殖能力的影响;Western blot检测白术内酯Ⅰ对TLR4及MyD88蛋白表达的作用。结果 qRT-PCR及免疫组化结果显示,相对于癌旁组织,TLR4及MyD88的mRNA和蛋白在肺癌组织中高表达(P<0.05);与对照组相比,白术内酯Ⅰ处理组A549细胞侵袭能力显著下降,增殖活力显著抑制(P<0.05);Western blot结果显示,白术内酯Ⅰ抑制TLR4及MyD88蛋白表达水平(P<0.05)。结论白术内酯Ⅰ通过抑制TLR4/MyD88通路抑制肺癌A549细胞侵袭转移。
Objective The objective of this study was to investigate the role and molecular mechanism of Atractylenolide Ⅰ in the progression of lung cancer.Methods qRT-qPCR and immunohistochemistry were used to detect the expression of TLR4 and MyD88 at levels of mRNA and protein in lung cancer and adjacent tissues.Transwell and MTT assays were used to detect effects of atractylenolide I(100 μM)on the invasion,migration and proliferation of A549 cells.Western blot was also used to detect the effect of atractylenolide I on the expression of TLR4 and MyD88 proteins.Results The expression of TLR4 and MyD88 at levels of mRNA and protein was highly expressed in lung cancer tissues when compared to adjacent tissues(P<0.05).Compared with the control group,the invasion ability of A549 cells in the Atractylenolide I group was significantly decreased,and the proliferative activity was inhibited(P<0.05).Atractylenolide I inhibited the expression of TLR4 and MyD88 protein in A549 cells(P<0.05).Conclusion Atractylenolide I inhibits the invasion and metastasis of lung cancer A549 cells by inhibiting the TLR4/MyD88 pathway.
Objective The objective of this study was to investigate the role and molecular mechanism of Atractylenolide Ⅰ in the progression of lung cancer.Methods qRT-qPCR and immunohistochemistry were used to detect the expression of TLR4 and MyD88 at levels of mRNA and protein in lung cancer and adjacent tissues.Transwell and MTT assays were used to detect effects of atractylenolide I(100 μM)on the invasion,migration and proliferation of A549 cells.Western blot was also used to detect the effect of atractylenolide I on the expression of TLR4 and MyD88 proteins.Results The expression of TLR4 and MyD88 at levels of mRNA and protein was highly expressed in lung cancer tissues when compared to adjacent tissues(P<0.05).Compared with the control group,the invasion ability of A549 cells in the Atractylenolide I group was significantly decreased,and the proliferative activity was inhibited(P<0.05).Atractylenolide I inhibited the expression of TLR4 and MyD88 protein in A549 cells(P<0.05).Conclusion Atractylenolide I inhibits the invasion and metastasis of lung cancer A549 cells by inhibiting the TLR4/MyD88 pathway.
引文
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13 Xiao Q,Zheng F,Wu JJ,et al.Activation of ERK and mutual regulation of Stat3 and SP1 contribute to inhibition of PDK1 expression by atractylenolide-1 in human lung cancer cells[J].Cell Physiol Biochem,2017,43(6):2353-2366.
14 Rogovskii VS.The linkage between inflammation and immune tolerance:interfering with inflammation in cancer[J].Curr Cancer Drug Targets,2017,17(4):325-332.
15 Xiao HC,Shu WL,Kui C.The role of toll-like receptor in inflammation and tumor immunity[J].Front Pharmacol,2018,9:878.
16 Zhu XX,Burfeind KG,Michaelis KA,et al.MyD88 signalling is critical in the development of pancreatic cancer cachexia[J].J Cachexia Sarcopenia Muscle,2019,10(2):378-390.
17 De Visser KE,Eichten A,Coussens LM.Paradoxical roles of the immune system during cancer development[J].Nature Reviews Cancer,2006,6(1):24-37.
18 Baniyash M.TCR zeta-chain downregulation:curtailing an excessive inflammatory immune response[J].Nat Rev Cancer,2004,4(9):675-687.
19 Rezvani K,Rouce R,Liu E,et al.Engineering natural killer cells for cancer immunotherapy[J].Mol Ther,2017,25(8):487-497.
2 Mittal D,Gubin MM,Schreiber RD,et al.New insights into cancer immunoediting and its three component phases-Elimination,equilibrium and escape[J].Curr Opin Immunol,2014,27:16-25.
3 Koliaraki V,Chalkidi N,Henriques A,et al.Innate sensing through mesenchymal TLR4/MyD88 signals promotes spontaneous intestinal tumorigenesis[J].Cell Rep,2019,26(3):536-545.
4 曲媛媛,徐向英,庄明,等.尼妥珠单抗对肺癌A549细胞系的体外放射增敏效应[J].实用肿瘤学杂志,2016,30(6):497-501.
5 Liu Y,Jia Z,Dong L,et al.A randomized pilot study of Atractylenolide I on gastric cancer cachexia patients[J].Evid Based Complement Alternat Med,2008,5(3):337-344.
6 Siegel RL,Miller KD,Jemal A.Cancer statistics,2019[J].CA Cancer J Clin,2019,69(1):7-34.
7 Pardoll DM.The blockade of immune checkpoints in cancer immunotherapy[J].Nat Rev Cancer,2012,12(4):252-264.
8 Dholaria B,Hammond W,Shreders A,et al.Emerging therapeutic agents for lung cancer[J].J Hematol Oncol,2016,9(1):138.
9 Ma L,Mao R,Shen K,et al.Atractylenolide I-mediated notch pathway inhibition attenuates gastric cancer stem cell traits[J].Biochem Biophys Res Commun,2014,450(1):353-359.
10 Huang HL,Lin TW,Huang YL,et al.Induction of apoptosis and differentiation by atractylenolide-1 isolated from atractylodes macrocephala in human leukemia cells[J].Bioorg Med Chem Lett,2016,26(8):1905-1909.
11 Ye Y,Chou GX,Wang H,et al.Effects of sesquiterpenes isolated from largehead atractylodes rhizome on growth,migration,and differentiation of B16 melanoma cells[J].Integr Cancer Ther,2011,10(1):92-100.
12 Qian X,Fang Z,Qing T,et al.Repression of PDK1-and LncRNA HOTAIR-mediated EZH2 gene expression contributes to the enhancement of atractylenolide 1 and erlotinib in the inhibition of human lung cancercells[J].Cell Physiol Biochem,2018,49(4):1615-1632.
13 Xiao Q,Zheng F,Wu JJ,et al.Activation of ERK and mutual regulation of Stat3 and SP1 contribute to inhibition of PDK1 expression by atractylenolide-1 in human lung cancer cells[J].Cell Physiol Biochem,2017,43(6):2353-2366.
14 Rogovskii VS.The linkage between inflammation and immune tolerance:interfering with inflammation in cancer[J].Curr Cancer Drug Targets,2017,17(4):325-332.
15 Xiao HC,Shu WL,Kui C.The role of toll-like receptor in inflammation and tumor immunity[J].Front Pharmacol,2018,9:878.
16 Zhu XX,Burfeind KG,Michaelis KA,et al.MyD88 signalling is critical in the development of pancreatic cancer cachexia[J].J Cachexia Sarcopenia Muscle,2019,10(2):378-390.
17 De Visser KE,Eichten A,Coussens LM.Paradoxical roles of the immune system during cancer development[J].Nature Reviews Cancer,2006,6(1):24-37.
18 Baniyash M.TCR zeta-chain downregulation:curtailing an excessive inflammatory immune response[J].Nat Rev Cancer,2004,4(9):675-687.
19 Rezvani K,Rouce R,Liu E,et al.Engineering natural killer cells for cancer immunotherapy[J].Mol Ther,2017,25(8):487-497.