Valproic acid(VPA) is a histone deacetylase inhibitor that has been an object of interest to clinicians for its promising potency in cancer therapy, as it induces apoptosis and differentiation, and enhances of chemoth...Valproic acid(VPA) is a histone deacetylase inhibitor that has been an object of interest to clinicians for its promising potency in cancer therapy, as it induces apoptosis and differentiation, and enhances of chemotherapy sensitivity. Esophageal squamous cell carcinoma(ESCC) is a malignant disease with growing incidence and low survival rate. Due to limited information on VPA activity in ESCC cells, we aimed to determine effects of VPA on chemotherapy responsiveness and expression of malignant markers in ESCC stem-like cells. Upon coadministration of non-toxic VPA + cisplatin(DDP), paclitaxel and 5-fluorouracil, viability of KYSE30 cells was assessed, and induced apoptosis was evaluated by DAPI staining, DNA laddering and flow cytometry. In addition, real time RT-PCR was performed to study changes in the expression of P21, CD44 and BMI-1 upon treatments. MTT test demonstrated that VPA significantly(P < 0.05) increased toxicity of DDP, which was confirmed by DNA laddering, flow cytometry analysis and significant(P < 0.05) overexpression of P21. Moreover, real time RT-PCR results indicated significant(P < 0.05) down regulation of CD44 and BMI-1 after VPA administration. Present attempt provided evidence, for the first time, that VPA not only improved responsiveness of esophageal stem-like cancer cells to DDP, also negatively regulated cancer stem cells markers in these cells.展开更多
目的:探讨甲硫氨酸代谢相关基因在食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)中的表达情况,研究甲硫氨酸和溶质载体家族43成员2(solute carrier family 43 member 2,SLC43A2)基因的表达对ESCC细胞迁移和侵袭的影响。方法...目的:探讨甲硫氨酸代谢相关基因在食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)中的表达情况,研究甲硫氨酸和溶质载体家族43成员2(solute carrier family 43 member 2,SLC43A2)基因的表达对ESCC细胞迁移和侵袭的影响。方法:利用生物信息学分析ESCC组织与邻近正常组织的甲硫氨酸代谢关键基因的表达情况,Kaplan-Meier生存曲线分析其表达在ESCC患者预后中的价值;聚类分析揭示不同表达模式对ESCC患者总体生存期和肿瘤微环境的影响;富集分析寻找ESCC中与甲硫氨酸和SLC43A2基因紧密关联的生物学进程;免疫组织化学染色检测ESCC组织和邻近正常组织中SLC43A2蛋白的表达情况;RT-qPCR和Western blot法检测干扰ESCC细胞SLC43A2效率;Transwell实验检测甲硫氨酸限制(methionine restriction)或干扰SLC43A2后ESCC细胞迁移和侵袭能力。结果:大部分甲硫氨酸代谢相关通路中的关键酶和SLC43A2基因在ESCC肿瘤组织中高表达(P<0.05),并且这些差异基因的表达对ESCC患者的预后具有显著影响(P<0.05);聚类分析显示高表达SLC43A2基因的A簇患者预后差,肿瘤微环境中免疫细胞种类和数量显著低于B簇;富集分析显示高表达SLC43A2基因组中上皮间充质转化(epithelial-mesenchymal transition,EMT)通路信号分子显著富集(P<0.01)。此外,限制甲硫氨酸或敲减SLC43A2可抑制ESCC细胞的迁移和侵袭(P<0.01)。结论:ESCC组织中甲硫氨酸代谢活跃,SLC43A2基因在ESCC组织中高表达与患者不良预后相关,SLC43A2促进ESCC细胞迁移和侵袭。展开更多
基金supported by a grant from Ferdowsi University of Mashhad(31546)
文摘Valproic acid(VPA) is a histone deacetylase inhibitor that has been an object of interest to clinicians for its promising potency in cancer therapy, as it induces apoptosis and differentiation, and enhances of chemotherapy sensitivity. Esophageal squamous cell carcinoma(ESCC) is a malignant disease with growing incidence and low survival rate. Due to limited information on VPA activity in ESCC cells, we aimed to determine effects of VPA on chemotherapy responsiveness and expression of malignant markers in ESCC stem-like cells. Upon coadministration of non-toxic VPA + cisplatin(DDP), paclitaxel and 5-fluorouracil, viability of KYSE30 cells was assessed, and induced apoptosis was evaluated by DAPI staining, DNA laddering and flow cytometry. In addition, real time RT-PCR was performed to study changes in the expression of P21, CD44 and BMI-1 upon treatments. MTT test demonstrated that VPA significantly(P < 0.05) increased toxicity of DDP, which was confirmed by DNA laddering, flow cytometry analysis and significant(P < 0.05) overexpression of P21. Moreover, real time RT-PCR results indicated significant(P < 0.05) down regulation of CD44 and BMI-1 after VPA administration. Present attempt provided evidence, for the first time, that VPA not only improved responsiveness of esophageal stem-like cancer cells to DDP, also negatively regulated cancer stem cells markers in these cells.
