染料木素抑制趋化因子受体4表达对膀胱癌T24细胞迁移和侵袭的影响机制
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摘要
目的探索染料木素是否能够抑制膀胱癌T24细胞的迁移和侵袭,以及这种抑制效应与趋化因子受体4(CXCR-4)及其下游信号通路的关系。方法实验设置对照组与3个染料木素组(10、20、40μg/mL组),未加染料木素的细胞作为对照组,染料木素组使用浓度为10、20、40μg/mL的染料木素培养液分别培养膀胱癌T24细胞。Transwell迁移小室检测染料木素对T24细胞迁移和侵袭能力的影响。蛋白印迹法检测CXCR-4及其下游信号分子Akt和mTOR的表达和活化水平。qPCR法检测CXCR-4与PI3K/Akt通路下游分子MMP-9和MMP-2的表达水平。结果染料木素显著抑制膀胱癌T24细胞的迁移和侵袭并呈剂量依赖性(P<0.05)。不同浓度的染料木素处理T24细胞后,蛋白印迹法和qPCR结果均显示CXCR-4表达显著下调(P<0.05)。进一步研究发现染料木素能够降低CXCR-4轴下游信号分子Akt和mTOR的活化水平(P<0.05),同时PI3K/Akt信号通路下游分子MMP-9和MMP-2的表达水平也显著降低(P<0.05)。结论染料木素通过抑制细胞CXCR-4及其下游信号通路的表达,从而抑制膀胱癌T24细胞的迁移和侵袭。
Objective To explore whether genistein inhibits the migration and invasion via the downregulation of CXCR-4 signaling in bladder cancer T24 cells.Methods In the experiment,bladder cancer T24 cells treated with 10,20,and 40 μg/mL group genistein were defined as the drug groups. The control cells were treated with vehicle. The effects of genistein on the migration and invasion of T24 cells were detected by Boyden chambers. The protein expression level of CXCR-4 and its down-stream signaling molecules Akt and mTOR were measured by Western blotting. The qPCR were used to detect the gene expression levels of CXCR-4 and the downstream molecules MMP-9 and MMP-2 of PI3K/Akt signaling.Results Genistein significantly inhibited the migration and invasion of bladder cancer T24 cells in a dose-dependent manner(P<0.05).After treated with indicated concentrations of genistein,the results of Western blotting and qPCR showed a significant down-regulation of CXCR-4 in T24 cells(P<0.05). Genistein down-regulated the activation levels of Akt and mTOR ina CXCR-4-dependent manner(P<0.05).After treatment with genistein,the expression levels of MMP-9 and MMP-2 were significantly decreased(P<0.05).Conclusion The genistein inhibits the migration and invasion of bladder cancer T24 cells through downregulating CXCR-4 signaling pathways.
Objective To explore whether genistein inhibits the migration and invasion via the downregulation of CXCR-4 signaling in bladder cancer T24 cells.Methods In the experiment,bladder cancer T24 cells treated with 10,20,and 40 μg/mL group genistein were defined as the drug groups. The control cells were treated with vehicle. The effects of genistein on the migration and invasion of T24 cells were detected by Boyden chambers. The protein expression level of CXCR-4 and its down-stream signaling molecules Akt and mTOR were measured by Western blotting. The qPCR were used to detect the gene expression levels of CXCR-4 and the downstream molecules MMP-9 and MMP-2 of PI3K/Akt signaling.Results Genistein significantly inhibited the migration and invasion of bladder cancer T24 cells in a dose-dependent manner(P<0.05).After treated with indicated concentrations of genistein,the results of Western blotting and qPCR showed a significant down-regulation of CXCR-4 in T24 cells(P<0.05). Genistein down-regulated the activation levels of Akt and mTOR ina CXCR-4-dependent manner(P<0.05).After treatment with genistein,the expression levels of MMP-9 and MMP-2 were significantly decreased(P<0.05).Conclusion The genistein inhibits the migration and invasion of bladder cancer T24 cells through downregulating CXCR-4 signaling pathways.
引文
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[15]陈伟.Kiss-1基因阻碍PI3K/AKT/NF-κB/MMP-9信号通路抑制结直肠癌细胞的增殖、迁移、侵袭和促进细胞凋亡的研究[D].福建医科大学,2016.
[2] Premo C,Apolo AB,Agarwal PK,et al.Trimodality therapy in bladder cancer:Who,what and when[J].Urol Clin North Am,2015,42(2):169-176.
[3] John BA,Said N.Insights from animal models of bladder cancer:recent advances,challenges,and opportunities[J].Oncotarget,2017,8(34):57766-57781.
[4] de la Parra C,Castillo-Pichardo L,Cruz-Collazo A,et al.Soy Isoflavone Genistein-Mediated Downregulation of miR-155 Contributes to the Anticancer Effects of Genistein[J]. Nutr Cancer,2016,68(1):154-164.
[5] Zhao Q,Zhao M,Parris AB,et al.Genistein targets the cancerous inhibitor of PP2A to induce growth inhibition and apoptosis in breast cancer cells[J].Int J Oncol,2016,49(3):1203-1210.
[6]蒋圣早,成启明.CXCR4和Ki-67表达与甲状腺癌预后的相关性研究[J].中国现代普通外科进展,2017,20(1):73-75.
[7]葛新,曹章,吕鹏威,等.miR-206抑制SDF-1/CXCR4信号活化诱导的乳腺癌细胞迁移和增殖[J].肿瘤基础与临床,2016,29(4):294-298.
[8] Liao YX,Fu ZZ,Zhou CH,et al.AMD3100 reduces CXCR4-mediated survival and metastasis of osteosarcoma by inhibiting JNK and Akt,but not p38 or Erk1/2,pathways in in vitro and mouse experiments[J].Oncol Rep,2015,34(1):33-42.
[9] Miyake M,Hori S,Morizawa Y,et al.CXCL1-Mediated Interaction of Cancer Cells with Tumor-Associated Macrophages and Cancer-Associated Fibroblasts Promotes Tumor Progression in Human Bladder Cancer[J].Neoplasia,2016,18(10):636-646.
[10] Spagnuolo C,Russo GL,Orhan IE,et al.Genistein and Cancer:Current Status,Challenges,and Future Directions[J].Adv Nutr,2015,6(4):408-419.
[11]周浩,张颖,李丹丹,等.SDF-1/CXCR4通路在干细胞心肌梗死治疗中的研究进展[J].中华临床医师杂志(电子版),2016(1):89-92.
[12] Huang X,Wu P,Huang F,et al.Baicalin attenuates chronic hypoxia-induced pulmonary hypertension via adenosine A2Areceptor-induced SDF-1/CXCR4/PI3K/AKT signaling[J].J Biomed Sci,2017,24(1):52.
[13]吴庆国,陈晔,江劲.温肾益气中药调节重症肺炎MMP-2和MMP-9表达的实验研究[J].中华全科医学,2016,14(7):1073-1076.
[14] Chen G,Yue Y,Qin J,et al.Plumbagin suppresses the migration and invasion of glioma cells via downregulation of MMP-2/9 expression and inaction of PI3K/Akt signaling pathway invitro[J].J Pharmacol Sci,2017,134(1):59-67.
[15]陈伟.Kiss-1基因阻碍PI3K/AKT/NF-κB/MMP-9信号通路抑制结直肠癌细胞的增殖、迁移、侵袭和促进细胞凋亡的研究[D].福建医科大学,2016.