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REG Iα 基因敲低通过细胞周期蛋白 D1/CDK4 通路和 BAX/BCL-2 通路增强结直肠癌细胞对氟尿嘧啶的敏感性。

Knockdown of REG Iα Enhances the Sensitivity to 5-Fluorouracil of Colorectal Cancer Cells via Cyclin D1/CDK4 Pathway and BAX/BCL-2 Pathways.

机构信息

1Department of Oncology, First Affiliated Hospital of Chongqing Medical University, Chongqing, China.

2Department of General Surgery, The Fifth People's Hospital of Chongqing, Chongqing, China.

出版信息

Cancer Biother Radiopharm. 2019 Aug;34(6):362-370. doi: 10.1089/cbr.2018.2746. Epub 2019 Apr 11.

Abstract

The reverse of chemoresistance and the improvement of sensitivity to chemotherapeutic agents of colorectal cancer cells have great clinical significance and the mechanism underlying the drug resistance is still unclear. REG Iα was reported to be upregulated in colorectal cancer tissues, but the roles of chemoresistance are still unclear. The expression of REG Iα in colorectal cancer cell lines was assessed by quantitative real-time polymerase chain reaction (Q-PCR). The expression of REG Iα in HCT116 and LOVO cells was knockdown by siRNA. The cell viability and IC (half maximal inhibitory concentration) values were analyzed by the CCK8 assay. The proportion of apoptosis and cell cycles were analyzed by flow cytometry. The migration potency of HCT116 and LOVO cells was analyzed by cell migration assay. The protein level of Cyclin D1, CDK4 (cyclin-dependent kinase 4), Bax and Bcl-2 were analyzed by western blot. Knockdown of REG Iα enhances the sensitivity to 5-Fu of colorectal cancer cells. REG Iα knockdown promoted the cell apoptosis of HCT116 and LOVO under the 5-Fu treatment. The cell migration and cycle of colorectal cancer cells was also inhibited by REG Iα knockdown. We also found that REG Iα knockdown induced cell cycle arrest and cell apoptosis by Cyclin D1/CDK4 pathway and BAX/BCL-2 pathways. Knockdown of REG Iα enhances the sensitivity to 5-Fu of colorectal cancer cells via cyclin D1/CDK4 pathway and BAX/BCL-2 pathways.

摘要

逆转结直肠癌细胞的化疗耐药性并提高对化疗药物的敏感性具有重要的临床意义,但其耐药机制尚不清楚。有报道称 REG Iα 在结直肠肿瘤组织中上调,但其耐药作用尚不清楚。通过实时定量聚合酶链反应(Q-PCR)评估结直肠癌细胞系中 REG Iα 的表达。用 siRNA 敲低 HCT116 和 LOVO 细胞中的 REG Iα 表达。通过 CCK8 检测分析细胞活力和 IC(半最大抑制浓度)值。通过流式细胞术分析细胞凋亡和细胞周期的比例。通过细胞迁移检测分析 HCT116 和 LOVO 细胞的迁移能力。通过 Western blot 分析细胞周期蛋白 D1、CDK4(细胞周期蛋白依赖性激酶 4)、Bax 和 Bcl-2 的蛋白水平。敲低 REG Iα 可增强结直肠癌细胞对 5-Fu 的敏感性。REG Iα 敲低可促进 HCT116 和 LOVO 在 5-Fu 处理下的细胞凋亡。REG Iα 敲低还抑制结直肠癌细胞的迁移和周期。我们还发现,REG Iα 敲低通过细胞周期蛋白 D1/CDK4 通路和 BAX/BCL-2 通路诱导细胞周期停滞和细胞凋亡。通过细胞周期蛋白 D1/CDK4 通路和 BAX/BCL-2 通路,敲低 REG Iα 可增强结直肠癌细胞对 5-Fu 的敏感性。

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