GOLPH3表达对人结肠癌5-氟尿嘧啶化疗耐药的影响

中华普通外科杂志 Pub Date : 2019-08-25 DOI:10.3760/CMA.J.ISSN.1007-631X.2019.08.016

Hai-Bin Zhuang, Chengzhi Qiu, Ming-Zhen Wang, Chunxiao Wang, Zhi-xiong Chen

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The protein expressions of GOLPH3, P-gp and β-catenin in HT29 cell were detected by Western blot. \n \n \nResults \nCompared with the control group, the expression level of GOLPH3 mRNA(1.000±0.078 vs. 0.147±0.021, t=12.296, P<0.01) and protein(1.003±0.235 vs. 0.077±0.399, t=6.723, P<0.01), the absorbance(A value) (1.000±0.082 vs. 0.769±0.086, t=3.885, P<0.01) and the tumorigenesis(430±36 vs. 297±21, t=5.492, P<0.01) in transfection group significantly reduced. The A value(0.803±0.101 vs. 1.050±0.140, t=2.855, P<0.05; 0.242±0.091 vs. 1.050±0.140, t=9.664, P<0.001) and the tumorigenesis(73±8 vs. 427±29, t=20.363, P<0.001; 305±22 vs .427±29, t=5.840, P<0.01) in experimental group 1 and experimental group 2 were significantly lower than that in the control group, while the A value(t=8.264, P<0.001) and the tumorigenesis(t=17.346, P<0.001) of experimental group 2 significantly decreased than that experimental group 1. 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引用次数: 0

摘要

目的探讨人结肠癌HT29细胞5-氟尿嘧啶(5-FU)化疗耐药与高尔基磷酸化蛋白3 (GOLPH3)表达的关系。方法将HT29细胞分为对照组、siRNA转染组、实验1组(30 μmol/L 5-FU)、实验2组(siRNA- golph3 + 30 μmol/L 5-FU)。采用RT-PCR和Western blot检测GOLPH3基因的沉默作用。采用MTT法和平板集落形成法分别测定HT29细胞的肿瘤增殖和肿瘤形成能力。Western blot检测HT29细胞中GOLPH3、P-gp和β-catenin蛋白的表达。结果与对照组比较，转染组GOLPH3 mRNA表达量(1.000±0.078比0.147±0.021,t=12.296, P<0.01)、蛋白表达量(1.003±0.235比0.077±0.399,t=6.723, P<0.01)、吸光度(A值)(1.000±0.082比0.769±0.086,t=3.885, P<0.01)、肿瘤发生(430±36比297±21,t=5.492, P<0.01)均显著降低。A值(0.803±0.101∶1.050±0.140)，t=2.855, P<0.05;(0.242±0.091∶1.050±0.140,t=9.664, P<0.001)和肿瘤发生(73±8∶427±29,t=20.363, P<0.001);实验1组和实验2组的A值(t=8.264, P<0.001)和肿瘤发生(t=17.346, P<0.001)均显著低于对照组(P<0.01)。与对照组(0.967±0.094,1.001±0.199,1.000±0.101)相比，实验1组GOLPH3、P-gp和β-catenin的表达量(3.634±0.574,2.424±0.261,2.324±0.418)显著上调(t=7.944, P<0.01;t = 7.520, P < 0.01;t=5.330, P<0.01)，而试验2组中这些蛋白的表达量(0.520±0.176,0.466±0.159,0.933±0.049)均显著低于试验1组(t=8.983, P<0.01;t = 11.106, P < 0.001;t = 5.724, P < 0.01)。结论GOLPH3的表达通过激活Wnt/β-catenin信号通路参与结肠癌HT29细胞对5-FU的耐药。关键词:结肠肿瘤;氟尿嘧啶;Wnt信号通路;22

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The effect of GOLPH3 expression on chemotherapeutic resistance to 5-Fluorouracil in human colon cancer cell lines

Objective To study the relationship between chemotherapeutic resistance to 5-fluorouracil (5-FU) and Golgi phosphoprotein 3 (GOLPH3) expression in human colon cancer cell line HT29. Methods HT29 cells were divided into four groups: control group, siRNA transfection group, experimental group 1(30 μmol/L 5-FU), and experimental group 2(siRNA-GOLPH3+ 30 μmol/L 5-FU). The silencing effect of GOLPH3 gene was detected by RT-PCR and Western blot. The tumor proliferation and formation ability of HT29 cell were measured respectively by MTT and plate colony formation assay. The protein expressions of GOLPH3, P-gp and β-catenin in HT29 cell were detected by Western blot. Results Compared with the control group, the expression level of GOLPH3 mRNA(1.000±0.078 vs. 0.147±0.021, t=12.296, P<0.01) and protein(1.003±0.235 vs. 0.077±0.399, t=6.723, P<0.01), the absorbance(A value) (1.000±0.082 vs. 0.769±0.086, t=3.885, P<0.01) and the tumorigenesis(430±36 vs. 297±21, t=5.492, P<0.01) in transfection group significantly reduced. The A value(0.803±0.101 vs. 1.050±0.140, t=2.855, P<0.05; 0.242±0.091 vs. 1.050±0.140, t=9.664, P<0.001) and the tumorigenesis(73±8 vs. 427±29, t=20.363, P<0.001; 305±22 vs .427±29, t=5.840, P<0.01) in experimental group 1 and experimental group 2 were significantly lower than that in the control group, while the A value(t=8.264, P<0.001) and the tumorigenesis(t=17.346, P<0.001) of experimental group 2 significantly decreased than that experimental group 1. Compared to the control group(0.967±0.094, 1.001±0.199, 1.000±0.101), the expressions of GOLPH3, P-gp and β-catenin in experimental group 1(3.634±0.574, 2.424±0.261, 2.324±0.418) significantly upregulated(t=7.944, P<0.01; t=7.520, P<0.01; t=5.330, P<0.01), while the expressions of these proteins in experimental group 2(0.520±0.176, 0.466±0.159, 0.933±0.049) were significantly lower than that in experimental group 1(t=8.983, P<0.01; t=11.106, P<0.001; t=5.724, P<0.01). Conclusion Expression of GOLPH3 is associated with resistance to 5-FU in colon cancer HT29 cell by activating Wnt/β-catenin signal pathway. Key words: Colonic neoplasms; Fluorouracil; Wnt signaling pathway; P-Glycoprotein

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中华普通外科杂志

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