Effect of GANT61 on Apoptosis of Esophageal Squamous Carcinoma Cells and Its Mechanism
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摘要:目的
研究Gli抑制剂GANT61诱导人食管鳞癌细胞OE21和KYSE-30凋亡的机制。
方法应用不同浓度（0、10、15 μmol/L）的GANT61处理OE21和KYSE-30细胞24 h后, Western blot观察GANT61作用于OE21和KYSE-30细胞后对Gli1、Gli2、Fas和Bcl-2蛋白表达的影响。流式细胞术观察细胞凋亡及Fas、Bcl-2蛋白的表达。
结果Western blot显示GANT61可以剂量依赖性显著下调OE21和KYSE-30细胞Gli1、Gli2和Bcl-2蛋白表达, 增加Fas蛋白表达。流式细胞术检测显示不同浓度GANT61作用24 h后, OE21和KYSE-30细胞表现出不同程度的细胞凋亡并且呈药物剂量依赖性, 与空白对照组相比差异均有统计学意义（P=0.000）；与空白对照组相比, OE21和KYSE-30细胞中Bcl-2蛋白表达水平显著降低且呈药物剂量依赖性, 差异均有统计学意义（P=0.000）； Fas蛋白表达水平显著增加且呈药物剂量依赖性, 差异均有统计学意义（P=0.000）。
结论GANT61明显诱导细胞凋亡, 其作用机制可能与其通过特异性抑制OE21和KYSE-30细胞中Gli1、Gli2表达从而调节Fas和Bcl-2蛋白表达有关。
Abstract:ObjectiveTo investigate the effect of GANT61 on the apoptosis of esophageal squamous carcinoma cells OE21 and KYSE-30, and to explore its possible mechanism.
MethodsAfter OE21 and KYSE-30 cell lines were treated with different concentrations（0, 10, 15μmol/L）of GANT61 for 24h, the protein expression of Gli1, Gli2, Fas and bcl-2 were detected by Western blot. Cell apoptosis, Fas and bcl-2 expression levels of OE21 and KYSE-30 cell lines were measured by flow cytometry.
ResultsWestern blot assay showed that GANT61 decreased Gli1, Gli2, Bcl-2 protein expression and increased Fas protein expression in a dose-dependent manner. Compared with DMSO group, the apoptotic rates of OE21 and KYSE-30 cell lines were significantly increased in a dose-dependent manner(P=0.000); the Bcl-2 protein expression levels were significantly decreased in a dose-dependent manner(P=0.000); the Fas protein expression levels were significantly increased in a dose-dependent manner(P=0.000).
ConclusionGANT61 could enhance the apoptotic rates of OE21 and KYSE-30 cells, and this effect may be associated with GANT61 decreasing Gli1 and Gli2 expression to regulate Fas and Bcl-2 protein expression levels in esophageal squamous carcinoma cells.
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Key words:
- Esophageal squamous carcinoma /
- Cell apoptosis /
- Gli /
- GANT61
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图 1 流式细胞术检测GANT61对OE21、KYSE-30细胞的诱导凋亡作用
Figure 1 Apoptosis of OE21 and KYSE-30 cells induced by GANT61 detected by FCM
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图 2 Western blot方法检测OE21细胞中蛋白表达水平
Figure 2 Protein expression of OE21 cells detected by Western blot
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图 3 Western blot方法检测KYSE-30细胞中蛋白表达水平
Figure 3 Protein expression of KYSE-30 cells detected by Western blot
表 1 流式细胞术检测不同浓度GANT61作用OE21、KYSE-30细胞24 h细胞中Bcl-2和Fas蛋白表达水平 (x±s, n=5)
Table 1 Bcl-2 and Fas protein expression levels in OE21, KYSE-30 cells after treated with different concentrations of GANT61 for 24h detected by flow cytometry (x±s, n=5)
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[1] Athar M, Li C, Kim AL, et al. Sonic hedgehog signaling in Basal cell nevus syndrome[J]. Cancer Res, 2014, 74(18): 4967-75. doi: 10.1158/0008-5472.CAN-14-1666
[1] Athar M, Li C, Kim AL, et al. Sonic hedgehog signaling in Basal cell nevus syndrome[J]. Cancer Res, 2014, 74(18): 4967-75. [2] Campbell V, Copland M. Hedgehog signaling in cancer stem cells: a focus on hematological cancers[J]. Stem Cells Cloning, 2015, 8: 27 -38. [2] Campbell V, Copland M. Hedgehog signaling in cancer stem cells: a focus on hematological cancers[J]. Stem Cells Cloning, 2015, 8: 27-38. http://cn.bing.com/academic/profile?id=1980798201&encoded=0&v=paper_preview&mkt=zh-cn
[3] Chandra V, Das T, Gulati P, et al. Hedgehog signaling pathway is active in GBM with GLI1 mRNA expression showing a single continuous distribution rather than discrete high/low clusters[J]. PLoS One, 2015, 10(3): e0116390. [3] Chandra V, Das T, Gulati P, et al. Hedgehog signaling pathway is active in GBM with GLI1 mRNA expression showing a single continuous distribution rather than discrete high/low clusters[J]. PLoS One, 2015, 10(3): e0116390. doi: 10.1371/journal.pone.0116390
[4] Felley-Bosco E, Opitz I, Meerang M. Hedgehog Signaling in Malignant Pleural Mesothelioma[J]. Genes (Basel), 2015, 6(3): 50 0-11. [4] Felley-Bosco E, Opitz I, Meerang M. Hedgehog Signaling in Malignant Pleural Mesothelioma[J]. Genes (Basel), 2015, 6(3): 500-11. http://cn.bing.com/academic/profile?id=1558724718&encoded=0&v=paper_preview&mkt=zh-cn
[5] Islam SS, Mokhtari RB, Noman AS, et al. Sonic hedgehog (Shh) signaling promotes tumorigenicity and stemness via activation of epithelial-to-mesenchymal transition (EMT) in bladder cancer[J]. Mol Carcinog, 2016, 55(5): 537-51. [5] Islam SS, Mokhtari RB, Noman AS, et al. Sonic hedgehog (Shh) signaling promotes tumorigenicity and stemness via activation of epithelial-to-mesenchymal transition (EMT) in bladder cancer[J]. Mol Carcinog, 2016, 55(5): 537-51. doi: 10.1002/mc.v55.5
[6] Behnsawy HM, Shigemura K, Meligy FY, et al. Possible role of sonic hedgehog and epithelial-mesenchymal transition in renal cell cancer progression[J]. Korean J Urol, 2013, 54(8): 547-54. [6] Behnsawy HM, Shigemura K, Meligy FY, et al. Possible role of sonic hedgehog and epithelial-mesenchymal transition in renal cell cancer progression[J]. Korean J Urol, 2013, 54(8): 547-54. doi: 10.4111/kju.2013.54.8.547
[7] Srivastava RK, Kaylani SZ, Edrees N, et al. GLI inhibitor GANT-61 diminishes embryonal and alveolar rhabdomyosarcoma growth by inhibiting Shh/AKT-mTOR axis[J]. Oncotarget, 2014, 5(23): 12151-65. doi: 10.18632/oncotarget
[7] Srivastava RK, Kaylani SZ, Edrees N, et al. GLI inhibitor GANT-61 diminishes embryonal and alveolar rhabdomyosarcoma growth by inhibiting Shh/AKT-mTOR axis[J]. Oncotarget, 2014, 5( 23): 12151-65. [8] Wang Y, Han C, Lu L, et al. Hedgehog signaling pathway regulates autophagy in human hepatocellular carcinoma cells[J]. Hepatology, 2013, 58(3): 995-1010. [8] Wang Y, Han C, Lu L, et al. Hedgehog signaling pathway regulates autophagy in human hepatocellular carcinoma cells[J]. Hepatology, 2013, 58(3): 995-1010. doi: 10.1002/hep.v58.3
[9] Wellbrock J, Latuske E, Kohler J, et al. Expression of Hedgehog Pathway Mediator GLI Represents a Negative Prognostic Marker in Human Acute Myeloid Leukemia and Its Inhibition Exerts Antileukemic Effects[J]. Clin Cancer Res, 2015, 21(10): 2388-98. doi: 10.1158/1078-0432.CCR-14-1059
[9] Wellbrock J, Latuske E, Kohler J, et al. Expression of Hedgehog Pathway Mediator GLI Represents a Negative Prognostic Marker in Human Acute Myeloid Leukemia and Its Inhibition Exerts Antileukemic Effects[J]. Clin Cancer Res, 2015, 21(10): 2388-98. [10] Regl G, Kasper M, Schnidar H, et al. Activation of the BCL2 promoter in response to Hedgehog/GLI signal transduction is predominantly mediated by GLI2[J]. Cancer Res, 2004, 64(21): 77 24-31. [10] Regl G, Kasper M, Schnidar H, et al. Activation of the BCL2 promoter in response to Hedgehog/GLI signal transduction is predominantly mediated by GLI2[J]. Cancer Res, 2004, 64(21): 7724-31. doi: 10.1158/0008-5472.CAN-04-1085
[11] Fu J, Rodova M, Roy SK, et al. GANT-61 inhibits pancreatic cancer stem cell growth in vitro and in NOD/SCID/IL2R gamma null mice xenograft[J]. Cancer Lett, 2013, 330(1): 22-32. [11] Fu J, Rodova M, Roy SK, et al. GANT-61 inhibits pancreatic cancer stem cell growth in vitro and in NOD/SCID/IL2R gamma null mice xenograft[J]. Cancer Lett, 2013, 330(1): 22-32. doi: 10.1016/j.canlet.2012.11.018
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