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    王腾, 张晖. KRT81在乳腺癌组织中的表达及其调控AKT/mTOR通路影响糖代谢抑制乳腺癌细胞增殖的作用[J]. 蚌埠医科大学学报, 2023, 48(11): 1488-1494. DOI: 10.13898/j.cnki.issn.1000-2200.2023.11.002
    引用本文: 王腾, 张晖. KRT81在乳腺癌组织中的表达及其调控AKT/mTOR通路影响糖代谢抑制乳腺癌细胞增殖的作用[J]. 蚌埠医科大学学报, 2023, 48(11): 1488-1494. DOI: 10.13898/j.cnki.issn.1000-2200.2023.11.002
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    WANG Teng, ZHANG Hui. Expression of KRT81 in breast cancer tissues and its inhibitory effect on proliferation of breast cancer cells by affecting glucose metabolism through regulating AKT/mTOR pathway[J]. Journal of Bengbu Medical University, 2023, 48(11): 1488-1494. DOI: 10.13898/j.cnki.issn.1000-2200.2023.11.002
    Citation: WANG Teng, ZHANG Hui. Expression of KRT81 in breast cancer tissues and its inhibitory effect on proliferation of breast cancer cells by affecting glucose metabolism through regulating AKT/mTOR pathway[J]. Journal of Bengbu Medical University, 2023, 48(11): 1488-1494. DOI: 10.13898/j.cnki.issn.1000-2200.2023.11.002
    shu

    KRT81在乳腺癌组织中的表达及其调控AKT/mTOR通路影响糖代谢抑制乳腺癌细胞增殖的作用

    Expression of KRT81 in breast cancer tissues and its inhibitory effect on proliferation of breast cancer cells by affecting glucose metabolism through regulating AKT/mTOR pathway

      摘要
    • 摘要:
      目的探究KRT81在乳腺癌组织中的表达及其调控AKT/mTOR通路影响糖代谢抑制乳腺癌细胞增殖的作用。
      方法收集100例乳腺癌病人的病理切片，免疫组织化学检测KRT81在乳腺癌中的表达，分析KRT81表达量与临床病理特征相关性。同时在MCF-7细胞中分别转染过表达质粒、对照质粒、对照敲低质粒、敲低质粒，记作oeKRT81组、NC组、Control组及si-KRT81组。通过集落克隆形成实验检测细胞增殖，流式细胞术检测凋亡细胞，蛋白印迹实验检测AKT/mTOR通路相关蛋白和细胞凋亡相关蛋白的表达。
      结果免疫组织化学检测结果显示，KRT81蛋白在癌组织中阳性表达明显低于癌旁正常组织（P < 0.01）。KRT81表达水平与Her2、TNM分期、远处转移和组织分级相关（P < 0.05~P < 0.01）。细胞集落克隆形成实验结果显示，oeKRT81组MCF-7细胞集落克隆数明显低于NC组，siKRT81组MCF-7细胞集落克隆数明显高于Control组（P < 0.01）。流式细胞术结果显示，与NC组相比，oeKRT81组MCF-7细胞凋亡率明显增加（P < 0.01）；与Control组相比，si-KRT81组MCF-7细胞凋亡率明显减少（P < 0.01）。蛋白印迹实验结果显示，oeKRT81组MCF-7细胞中Bax蛋白水平明显高于NC组，Bcl2、GLUT1、LDH、p-AKT、p-mTOR蛋白水平明显低于NC组（P < 0.01）；si-KRT81组Bax蛋白水平明显低于Control组，Bcl2、GLUT1、LDH、p-AKT、p-mTOR蛋白水平明显Control组（P < 0.01）。
      结论KRT81低表达与乳腺癌的不良预后结局相关，其可能通过抑制AKT/mTOR通路影响糖代谢抑制乳腺癌细胞增殖。

       

      Abstract:
      ObjectiveTo investigate the expression of KRT81 in breast cancer tissues and its inhibitory effect on proliferation of breast cancer cells by affecting glucose metabolism through regulating AKT/mTOR pathway.
      MethodsThe pathological sections of 100 patients with breast cancer were collected, the expression of KRT81 in breast cancer was detected by immunohistochemistry, and the correlation between KRT81 expression level and clinical pathological characteristics was analyzed.Simultaneously, overexpression plasmids, control plasmids, control knockdown plasmids, and knockdown plasmids were transfected into MCF-7 cells, which were recorded as oeKRT81 group, NC group, Control group, and si-KRT81 group.The cell proliferation was detected by colony formation assay, apoptosis was determined by flow cytometry, and the expression of AKT/mTOR pathway-related proteins and apoptosis-related proteins were detected by Western blotting.
      ResultsThe results of immunohistochemistry showed that the positive expression of KRT81 protein in cancer tissues was significantly lower than that in normal adjacent tissues (P < 0.01).The expression level of KRT81 was correlated with Her2, TNM staging, distant metastasis, and tissue grading (P < 0.05 to P < 0.01).The results of the colony formation assay showed that the number of MCF-7 cell colonies in the oeKRT81 group was significantly lower than that in the NC group, while the number of MCF-7 cell colonies in the siKRT81 group was significantly higher than that in the Control group (P < 0.01).The results of flow cytometry showed that compared with the NC group, the apoptosis rate of MCF-7 cells in the oeKRT81 group was significantly increased (P < 0.01); compared with the Control group, the apoptosis rate of MCF-7 cells in the si-KRT81 group was significantly reduced (P < 0.01).The results of Western blotting indicated that the level of Bax protein in MCF-7 cells in the oeKRT81 group were significantly higher than that in the NC group, while the levels of Bcl2, GLUT1, LDH, p-AKT, and p-mTOR proteins were significantly lower than those in the NC group (P < 0.01);the level of Bax protein in the si-KRT81 group was significantly lower than that in the Control group, while the levels of Bcl2, GLUT1, LDH, p-AKT, and p-mTOR proteins were significantly higher than those in the Control group (P < 0.01).
      ConclusionsThe low expression of KRT81 is related to poor prognosis of breast cancer, which may inhibit the proliferation of breast cancer cells by affecting glucose metabolism through inhibiting AKT/mTOR pathway.

       

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