Combretastatin A4 phosphate on proliferation, migration and signal pathway of U87 cells
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摘要:
目的 研究药物康普瑞汀磷酸钠（CA4P）对人胶质瘤细胞U87细胞增殖迁移的影响以及相关信号通路。 方法 体外培养U87细胞并用不同浓度的CA4P处理，CCK8方法检测不同药物浓度对细胞活性的影响，划痕和transwell实验检测不同药物浓度对U87细胞迁移能力的影响，Western blot检测不同浓度处理后U87细胞中E-Cadherin蛋白。Path Scan对相关激活的通路进行检测。 结果 CA4P能够明显抑制体外U87细胞增殖。划痕和transwell实验表明CA4P能够在体外抑制U87细胞的迁移能力，CA4P浓度为50 nmol/L和100 nmol/L能显著抑制迁移（P<0.001）。不同浓度的CA4P作用细胞24 h后，E-Cadherin的蛋白水平随着CA4P的浓度增加而增高。Path Scan结果表明，细胞内信号通路Akt, Bad, SAPK/JNK, S6 ribosomal Protein被激活。 结论 CA4P能够抑制人U87细胞的增殖和迁移，并且可能与Akt, Bad, SAPK/JNK的上调和S6 ribosomal Protein下调有关。 Abstract:Objective To explore the effect of drug combretast sodium phosphate (CA4P) on the proliferation and migration of human glioma U87 cells and related signaling pathways. Methods U87 cells were cultured in vitro and treated with different concentrations of CA4P. The effects of different concentrations of drugs on cell viability were detected by CCK8 assay. Scratches and transwell assays were performed to examine the effect of different drug concentrations on migration of U87 cells. Western blot was used to detect E-Cadherin protein in U87 cells treated with different concentrations. Path Scan to detect the activation of the pathway. Results The effect of CA4P could inhibit the proliferation of U87 cells in vitro. Scratches and transwell assays showed that CA4P inhibited the migration of U87 cells in vitro. The concentration of CA4P of 50 nM and 100 nM significantly inhibited the migration. After treated with different concentrations of CA4P for 24 h, the protein level of E-Cadherin increased with the increase of CA4P concentration. Path Scan results showed that the intracellular signaling pathways Akt, Bad, SAPK/JNK and S6 ribosomal Protein were activated. Conclusion CA4P can inhibit the proliferation and migration of human U87 cells, and may be related to the upregulation of Akt, Bad, SAPK/JNK and the downregulation of S6 ribosomal protein. -
Key words:
- CA4P /
- glioma /
- proliferation /
- migration /
- signal pathway
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图 1 CA4P抑制U87细胞的增殖
A: U87细胞分别用0、6.25、12.5、25、50、100 nmol/L CA4P刺激, CCK-8分析CA4P对U87细胞活性的影响; B: CA4P作用细胞24 h的统计图，与对照组相比CA4P在24 h内不影响细胞活性，差异没有统计学意义（P>0.05）.
图 2 CA4P抑制U87细胞的迁移
A, B: 分析0、6.25、12.5、25、50、100 nmol/LCA4P对U87细胞的迁移能力影响; C: 差异具有显著性，**P<0.001.
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[1] Das S, Marsden PA. Angiogenesis in glioblastoma[J]. N Engl J Med, 2013, 369(16): 1561-3 [2] Alexander BM, Cloughesy TF. Adult glioblastoma[J]. J Clin Oncol, 2017, 35(21): 2402-9 [3] Sampetrean O, Saya H. Modeling phenotypes of malignant gliomas[J]. Cancer Sci, 2017, 32(10): 1111-7 [4] Taal W, Bromberg JE. chemotherapy in glioma[J]. CNS Oncol, 2015, 4(3): 179-92 [5] Stavrovskaya AA, Shushanov SS, Rybalkina EY. Problems of glioblastoma multiforme drug resistance[J]. Biochemistry, 2016, 81(2): 91-100 [6] Fan CH, Liu WL, Cao H, et al. O6-methylguanine DNA methyltransferase as a promising target for the treatment of temozolomide-resistant gliomas[J]. Cell Death Dis, 2013, 4(10): e876-81 [7] Yi GZ, Liu YW, Xiang W, et al. Akt and beta-catenin contribute to TMZ resistance and EMT of MGMT negative malignant glioma cell line[J]. J Neurol Sci, 2016, 367(12): 101-6 [8] West CM, Price P. Combretastatin a4 phosphate[J]. Anticancer Drugs, 2004, 15(3): 179-87 [9] Chase DM, Chaplin DJ, Monk BJ. The development and use of vascular targeted therapy in ovarian cancer[J]. Gynecol Oncol, 2017, 145(2): 393-406 [10] Garon EB, Neidhart JD, Gabrail NY, et al. A randomized Phase II trial of the tumor vascular disrupting agent CACA4P (fosbretabulin tromethamine) with carboplatin, paclitaxel, and bevacizumab in advanced nonsquamous non-small-cell lung cancer[J]. Onco Targets Ther, 2016, 9(10): 7275-83 [11] Nathan P, Zweifel M, Padhani AR, et al. Phase I trial of combretastatin A4 phosphate (CA4P) in combination with bevacizumab in patients with advanced cancer[J]. Clin Cancer Res, 2012, 18(12): 3428-39 [12] Liu L, Mason RP, Gimi B. Dynamic bioluminescence and fluorescence imaging of the effects of the antivascular agent Combretastatin-A4P (CA4P) on brain tumor xenografts[J]. Cancer Lett, 2015, 356(2B): 462-9 [13] Vincent L. Combretastatin a4 phosphate induces rapid regression of tumor neovessels and growth through interference with vascular endothelial-cadherin signaling[J]. J Clin Investig, 2005, 115(11): 2992-3006 [14] 陈 灿, 牛 犇, 包 旭, 等. CA4P抗肿瘤作用的实验研究[J]. 华西药学杂志, 2009, 24(6): 609-11 [15] 王 艳, 朱丽媛, 刘 畅, 等. Combretastatin A4 Phosphate对人晶状体上皮细胞增殖及迁移的影响[J]. 中山大学学报: 医学科学版, 2012, 33(1): 8-15 [16] 牛 犇, 方素华, 包 旭, 等. 康普瑞丁磷酸二钠在体内外的抗肿瘤作用[J]. 华西药学杂志, 2007, 22(6): 632-5 [17] Feitelson MA, Arzumanyan A, Kulathinal RJ, et al. Sustained proliferation in cancer: Mechanisms and novel therapeutic targets[J]. Semin Cancer Biol, 2015, 35(S): S25-54 [18] Harris AL. Hypoxia-A key regulatory factor in tumour growth[J]. Nat Rev Cancer, 2002, 2(1): 38-47 [19] Chen HF, Ma RR, He JY, et al. Protocadherin 7 inhibits cell migration and invasion through E-cadherin in gastric cancer[J]. Tumor Biol, 2017, 39(4): 568835941-9 [20] Zhang L, Wang GY, Wang L, et al. Valproic acid inhibits prostate cancer cell migration by up-regulating E-cadherin expression[J]. Pharmazie, 2011, 66(8): 614-8 [21] Petit I, Karajannis MA, Vincent L, et al. The microtubule-targeting agent CA4P regresses leukemic xenografts by disrupting interaction with vascular cells and mitochondrial-dependent cell death[J]. Blood, 2008, 111(4): 1951-61 [22] Izumi Y, Aoshima K, Hoshino Y, et al. Effects of combretastatin A-4 phosphate on canine normal and tumor tissue-derived endothelial cells[J]. Res Vet Sci, 2017, 112(6): 222-8 [23] Chen BJ, Zhang W, Gao J, et al. Downregulation of ribosomal protein S6 inhibits the growth of non-small cell lung cancer by inducing cell cycle arrest, rather than apoptosis[J]. Cancer Lett, 2014, 354(2): 378-89 [24] Lin A, Piao HL, Zhuang L, et al. FoxO transcription factors promote AKT Ser473 phosphorylation and renal tumor growth in response to pharmacologic inhibition of the PI3K-AKT pathway[J]. Cancer Res, 2014, 74(6): 1682-93 [25] Xie Q, Sun Z, Chen M, et al. IL-8 up-regulates proliferative angiogenesis in ischemic myocardium in rabbits through phosphorylation of Akt/GSK-3β(ser9)dependent pathways[J]. Int J Clin Exp Med, 2015, 8(8): 12498-507 [26] Zhang ZW, Zhang HQ, Chen SY, et al. Dihydromyricetin induces mitochondria-mediated apoptosis in HepG2 cells through down-regulation of the Akt/Bad pathway[J]. Nutr Res, 2017, 38(5): 27-33 [27] Zheng GY, Chen XC, Du J. Inhibitory action of propyl gallate on the activation of SAPK/JNK and p38MAPK induced by cerebral ischem ia-reperfusion in rats[J]. Acta Pharm Sin, 2006, 41(6): 548-54 [28] Cicenas J. The potential role of Akt phosphorylation in human cancers[J]. Int J Biol Markers, 2008, 23(1): 1-9 -
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