PDF(Pubmed)
Abstract：
UNASSIGNED: Colorectal cancer (CRC) is one of the most frequently diagnosed cancers. In many cases, the poor prognosis of advanced CRC is associated with resistance to treatment with chemotherapeutic drugs such as 5-Fluorouracil (5-FU). The epithelial-to-mesenchymal transition (EMT) and dysregulation in protein methylation are two mechanisms associated with chemoresistance in many cancers. This study looked into the effect of 5-FU dose escalation on EMT and protein methylation in CRC.
UNASSIGNED: HCT-116, Caco-2, and DLD-1 CRC cell lines were exposed to dose escalation treatment of 5-FU. The motility and invasive potentials of the cells before and after treatment with 5-FU were investigated through wound healing and invasion assays. This was followed by a Western blot which analyzed the protein expressions of the epithelial marker E-cadherin, mesenchymal marker vimentin, and the EMT transcription factor (EMT-TF), the snail family transcriptional repressor 1 (Snail) in the parental and desensitized cells. Western blotting was also conducted to study the protein expressions of the protein methyltransferases (PMTs), Euchromatic histone lysine methyltransferase 2 (EHMT2/G9A), protein arginine methyltransferase (PRMT5), and SET domain containing 7/9 (SETD7/9) along with the global lysine and arginine methylation profiles.
UNASSIGNED: The dose escalation method generated 5-FU desensitized CRC cells with distinct morphological features and increased tolerance to high doses of 5-FU. The 5-FU desensitized cells experienced a decrease in migration and invasion when compared to the parental cells. This was reflected in the observed reduction in E-cadherin, vimentin, and Snail in the desensitized cell lines. Additionally, the protein expressions of EHMT2/G9A, PRMT5, and SETD7/9 also decreased in the desensitized cells and global protein lysine and arginine methylation became dysregulated with 5-FU treatment.
UNASSIGNED: This study showed that continuous, dose-escalation treatment of 5-FU in CRC cells generated 5-FU desensitized cancer cells that seemed to be less aggressive than parental cells.
UNASSIGNED: HCT-116, Caco-2, and DLD-1 CRC cell lines were exposed to dose escalation treatment of 5-FU. The motility and invasive potentials of the cells before and after treatment with 5-FU were investigated through wound healing and invasion assays. This was followed by a Western blot which analyzed the protein expressions of the epithelial marker E-cadherin, mesenchymal marker vimentin, and the EMT transcription factor (EMT-TF), the snail family transcriptional repressor 1 (Snail) in the parental and desensitized cells. Western blotting was also conducted to study the protein expressions of the protein methyltransferases (PMTs), Euchromatic histone lysine methyltransferase 2 (EHMT2/G9A), protein arginine methyltransferase (PRMT5), and SET domain containing 7/9 (SETD7/9) along with the global lysine and arginine methylation profiles.
UNASSIGNED: The dose escalation method generated 5-FU desensitized CRC cells with distinct morphological features and increased tolerance to high doses of 5-FU. The 5-FU desensitized cells experienced a decrease in migration and invasion when compared to the parental cells. This was reflected in the observed reduction in E-cadherin, vimentin, and Snail in the desensitized cell lines. Additionally, the protein expressions of EHMT2/G9A, PRMT5, and SETD7/9 also decreased in the desensitized cells and global protein lysine and arginine methylation became dysregulated with 5-FU treatment.
UNASSIGNED: This study showed that continuous, dose-escalation treatment of 5-FU in CRC cells generated 5-FU desensitized cancer cells that seemed to be less aggressive than parental cells.
摘要：
结直肠癌(CRC)是最常见的诊断癌症之一。在许多情况下,晚期CRC的不良预后与对5-氟尿嘧啶(5-FU)等化疗药物治疗耐药相关.上皮间质转化（EMT）和蛋白质甲基化失调是许多癌症中与化学耐药相关的两种机制。本研究观察了5-FU剂量递增对CRC中EMT和蛋白质甲基化的影响。
■将HCT-116、Caco-2和DLD-1CRC细胞系暴露于5-FU的剂量递增处理。通过伤口愈合和侵袭试验研究了5-FU治疗前后细胞的运动性和侵袭潜力。然后进行蛋白质印迹，分析上皮标记E-cadherin的蛋白质表达，间充质标记物波形蛋白，和EMT转录因子（EMT-TF），亲本和脱敏细胞中的蜗牛家族转录抑制因子1（Snail）。还进行了蛋白质印迹以研究蛋白质甲基转移酶（PMT）的蛋白质表达，优色组蛋白赖氨酸甲基转移酶2（EHMT2/G9A），蛋白质精氨酸甲基转移酶（PRMT5），和SET结构域包含7/9（SETD7/9）以及全局赖氨酸和精氨酸甲基化谱。
■剂量递增方法产生的5-FU脱敏CRC细胞具有明显的形态学特征和对高剂量5-FU的增加的耐受性。当与亲代细胞相比时,5-FU脱敏的细胞经历迁移和侵袭的减少。这反映在观察到的E-cadherin减少，波形蛋白,和蜗牛在脱敏细胞系中。此外,EHMT2/G9A的蛋白表达，PRMT5和SETD7/9在脱敏细胞中也降低，并且5-FU处理后，整体蛋白赖氨酸和精氨酸甲基化变得失调。
■这项研究表明，5-FU在CRC细胞中的剂量递增处理产生了5-FU脱敏的癌细胞,其似乎不如亲本细胞具有侵袭性.
■将HCT-116、Caco-2和DLD-1CRC细胞系暴露于5-FU的剂量递增处理。通过伤口愈合和侵袭试验研究了5-FU治疗前后细胞的运动性和侵袭潜力。然后进行蛋白质印迹，分析上皮标记E-cadherin的蛋白质表达，间充质标记物波形蛋白，和EMT转录因子（EMT-TF），亲本和脱敏细胞中的蜗牛家族转录抑制因子1（Snail）。还进行了蛋白质印迹以研究蛋白质甲基转移酶（PMT）的蛋白质表达，优色组蛋白赖氨酸甲基转移酶2（EHMT2/G9A），蛋白质精氨酸甲基转移酶（PRMT5），和SET结构域包含7/9（SETD7/9）以及全局赖氨酸和精氨酸甲基化谱。
■剂量递增方法产生的5-FU脱敏CRC细胞具有明显的形态学特征和对高剂量5-FU的增加的耐受性。当与亲代细胞相比时,5-FU脱敏的细胞经历迁移和侵袭的减少。这反映在观察到的E-cadherin减少，波形蛋白,和蜗牛在脱敏细胞系中。此外,EHMT2/G9A的蛋白表达，PRMT5和SETD7/9在脱敏细胞中也降低，并且5-FU处理后，整体蛋白赖氨酸和精氨酸甲基化变得失调。
■这项研究表明，5-FU在CRC细胞中的剂量递增处理产生了5-FU脱敏的癌细胞,其似乎不如亲本细胞具有侵袭性.
