硫酸右旋糖苷抑制人胃癌细胞增殖、迁移及相关因子表达的作用
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摘要
目的探讨硫酸右旋糖苷(DS)对人胃癌细胞(BGC-823)增殖、侵袭与迁移能力的作用及其可能的分子机制。方法模拟胃癌缺氧微环境,在低氧条件下培养人胃癌细胞(BGC-823),设立对照组与实验组,分别予以磷酸缓冲盐溶液(PBS)与DS处理,采用平板克隆形成实验观察DS对细胞增殖能力的影响;Transwell侵袭与Transwell迁移实验分别观察DS对细胞侵袭与迁移能力的影响;免疫细胞荧光、Western Blot与Q-PCR实验检测经低氧条件培养不同时间(2、8、12、24 h)后细胞中转录因子NF-E2相关因子2(Nrf2)、血红素氧合酶-1(HO-1)的表达水平。结果克隆形成实验结果显示,实验组与对照组相比细胞克隆形成数量明显减少(P <0.05);Transwell侵袭与迁移实验结果显示,实验组与对照组相比侵袭与迁移的细胞数量均明显减少(P <0.05);免疫细胞荧光、Western Blot与Q-PCR实验结果显示,不同时间点实验组Nrf2、HO-1的表达水平比对照组明显降低(P <0.05)。结论 DS能够抑制人胃癌细胞(BGC-823)增殖、侵袭与迁移的能力,其机制可能与下调Nrf2、HO-1的表达水平有关。
Objective To investigate the effect of dextran sulfate(DS)on the proliferation,invasion andmigration of human gastric cancer BGC-823 cells and its possible molecular mechanism. Methods To simulatethe hypoxic microenvironment of gastric cancer,human gastric cancer BGC-823 cells were divided into controlgroup and experimental group,phosphate buffered saline(PBS)and DS were added respectively under hypoxicconditions. The effect of DS on cell proliferation was observed by plate colony formation assay. The effects of DS oncell invasion and migration were observed by Transwell invasion and migration assay. The expression levels of nuclearfactor-erythroid 2-related factor 2(Nrf2)and heme oxygenase 1(HO-1) in cells were detected by immunofluores-cence,Western Blot and Q-PCR after incubation for different time(2,8,12,24 h). Results The results ofplate colony formation assay showed that the number of cell clones in the experimental group was significantlyreduced,compared with the control group. Transwell invasion and migration assay showed that the number of cellsinvading and migrating was significantly reduced in the experimental group compared with the control group. Theresults of immunofluorescence,Western Blot and Q-PCR showed that the expression levels of Nrf2 and HO-1 inexperimental group were significantly lower than those in control group at different time points. Conclusion DScan inhibit the proliferation,invasion and migration of human gastric cancer BGC-823 cells,and its mechanismmay be related to down-regulation of Nrf2 and HO-1 expression levels.
Objective To investigate the effect of dextran sulfate(DS)on the proliferation,invasion andmigration of human gastric cancer BGC-823 cells and its possible molecular mechanism. Methods To simulatethe hypoxic microenvironment of gastric cancer,human gastric cancer BGC-823 cells were divided into controlgroup and experimental group,phosphate buffered saline(PBS)and DS were added respectively under hypoxicconditions. The effect of DS on cell proliferation was observed by plate colony formation assay. The effects of DS oncell invasion and migration were observed by Transwell invasion and migration assay. The expression levels of nuclearfactor-erythroid 2-related factor 2(Nrf2)and heme oxygenase 1(HO-1) in cells were detected by immunofluores-cence,Western Blot and Q-PCR after incubation for different time(2,8,12,24 h). Results The results ofplate colony formation assay showed that the number of cell clones in the experimental group was significantlyreduced,compared with the control group. Transwell invasion and migration assay showed that the number of cellsinvading and migrating was significantly reduced in the experimental group compared with the control group. Theresults of immunofluorescence,Western Blot and Q-PCR showed that the expression levels of Nrf2 and HO-1 inexperimental group were significantly lower than those in control group at different time points. Conclusion DScan inhibit the proliferation,invasion and migration of human gastric cancer BGC-823 cells,and its mechanismmay be related to down-regulation of Nrf2 and HO-1 expression levels.
引文
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[2]LI W,NG J M,WONG C C,et al.Molecular alterations of cancer cell and tumour microenvironment in metastatic gastric cancer[J].Oncogene,2018,37(36):4903-4920.
[3]XU Y,JIN X,HUANG Y,et al.Dextran sulfate inhibition on human gastric cancer cells invasion,migration and epithelialmesenchymal transformation[J].Oncol Lett,2018,16(4):5041-5049.
[4]XU Y,HUANG Y,WANG H,et al.Inhibition of the peritoneal metastasis of human gastric cancer cells by dextran sulphate in vivo and in vitro[J].Oncol Lett,2016,11(4):2384-2390.
[5]高文卫,金秀,田润玲,等.硫酸右旋糖酐抑制人胃癌细胞增殖、侵袭和迁移的作用[J].实用医学杂志,2018,34(21):3528-3532.
[6]FURFARO A L,TRAVERSO N,DOMENICOTTI C,et al.The Nrf2/HO-1 axis in cancer cell growth and chemoresistance[J].Oxid Med Cell Longev,2016,2016(20):1-14.
[7]SONG Z,WU Y,YANG J,et al.Progress in the treatment of advanced gastric cancer[J].Tumour Biol,2017,39(7):1010428317714626.
[8]NAJAFI M,GORADEL N H,FARHOOD B,et al.Tumor microenvironment:Interactions and therapy[J].J Cell Physiol,2019,234(5):5700-5721.
[9]SUN F,FENG M,GUAN W.Mechanisms of peritoneal dissemination in gastric cancer[J].Oncol Lett,2017,14(6):6991-6998.
[10]SOVA M,SASO L.Design and development of Nrf2 modulators for cancer chemoprevention and therapy:a review[J].Drug Des Devel Ther,2018,25(12):3181-3197.
[11]ZELENKA J,KONCOSOVA M,RUML T.Targeting of stress response pathways in the prevention and treatment of cancer[J].Biotechnol Adv,2018,36(3):583-602.
[12]NITTI M,PIRAS S,MARINARI U M,et al.HO-1 Induction in cancer progression:a matter of cell adaptation[J].Antioxidants(Basel),2017,6(2).pii:E29
[13]MENEGON S,COLUMBANO A,GIORDANO S.The dual roles of nrf2 in cancer[J].Trends Mol Med,2016,22(7):578-593.
[14]TAGUCHI K,MOTOHASHI H,YAMAMOTO M.Molecular mechanisms of the Keap1-Nrf2 pathway in stress response and cancer evolution[J].Genes Cells,2011,16(2):123-140.
[15]KIM T H,HUR E G,KANG S J,et al.NRF2 blockade suppresses colon tumor angiogenesis by inhibiting hypoxia-induced activation of HIF-1alpha[J].Cancer Res,2011,71(6):2260-2275.
[16]LIU R,PENG J,WANG H,et al.Oxysophocarpine retards the growth and metastasis of oral squamous cell carcinoma by targeting the Nrf2/HO-1 axis[J].Cell Physiol Biochem,2018,49(5):1717-1733.
[17]RACHAKONDA G,SEKHAR K R,JOWHAR D,et al.Increased cell migration and plasticity in Nrf2-deficient cancer cell lines[J].Oncogene,2010,29(25):3703-3714.
[18]PAN H,WANG H,ZHU L,et al.The role of Nrf2 in migration and invasion of human glioma cell U251[J].World Neurosurg,2013,80(3-4):363-370
[19]TSAI J R,WANG H M,LIU P L,et al.High expression of heme oxygenase-1 is associated with tumor invasiveness and poor clinical outcome in non-small cell lung cancer patients[J].Cell Oncol(Dordr),2012,35(6):461-471.
[20]MENG D,WANG X,CHANG Q,et al.Arsenic promotes angiogenesis in vitro via a heme oxygenase-1-dependent mechanism[J].Toxicol Appl Pharmacol,2010,244(3):291-299