结直肠癌细胞中NF-κB活化与TRAF6、CCR5及PTEN/PI3K通路的关系及作用
作者:
作者单位:

1.宁夏医科大学总医院,外科学研究室,宁夏 银川 750004;2.宁夏医科大学总医院,结直肠外科,宁夏 银川 750004;3.宁夏医科大学,宁夏 银川 750004

作者简介:

何亚琴,宁夏医科大学总医院助理研究员,主要从事肠道免疫方面的研究。

通信作者:

谢小亮,Email: xxl811001@126.com

基金项目:

宁夏回族自治区“十三五”重点研发计划基金资助项目(2016BZ02);宁夏回族自治区重点研发计划基金资助项目(2021BEG03086) 。


Association of NF-κB activation with TRAF6, CCR5 and PTEN/PI3K pathway and its role in colorectal cancer cells
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Affiliation:

1.Department of Surgical Research, General Hospital of Ningxia Medical University, Yinchuan 750004, China;2.Department of Colorectal Surgery, General Hospital of Ningxia Medical University, Yinchuan 750004, China;3.Ningxia Medical University, Yinchuan 750004, China

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    摘要:

    背景与目的 NF-κB的活化在多种恶性肿瘤的发生、发展中起了重要的作用。研究发现,趋化因子受体5(CCR5)、肿瘤坏死因子受体相关因子6(TRAF6)以及PTEN/PI3K通路相关蛋白均在恶性肿瘤中异常表达。因此,本研究探讨以上分子在结直肠癌细胞中的作用及相互关系。方法 分别用Western blot、CCK-8实验、Transwell法检测结直肠癌HT29和SW480细胞经Maraviroc(CCR5抑制剂)、MG132(TRAF6抑制剂)和NF-BAY(NF-κB抑制剂)处理后,各蛋白表达的变化,以及增殖、迁移、侵袭能力的变化。结果 在两种结直肠癌细胞中,抑制CCR5蛋白后,PI3K的表达降低,PTEN表达升高(均P<0.05),TRAF6和NF-κB表达无明显变化(均P>0.05);抑制TRAF6蛋白后,PI3K和CCR5表达降低,PTEN表达升高(均P<0.05),NF-κB的表达无明显变化(均P>0.05);抑制NF-κB表达后,CCR5、TRAF6和PI3K表达降低,PTEN表达升高(均P<0.05)。三种抑制剂均可明显降低两种结直肠癌细胞的增殖、迁移和侵袭能力(均P<0.05)。结论 结直肠癌细胞中存在NF-κB异常活化,后者可能通过上调TRAF6与CCR5的表达,抑制抑癌分子PTEN的活性,从而导致促癌分子PI3K及其通路的活性升高。

    Abstract:

    Background and Aims The activation of NF-κB plays a crucial role in development and progression of various malignant tumors. Studies demonstrated abnormal expressions in tumor necrosis factor receptor-associated factor 6 (TRAF6) and proteins associated with PTEN/PI3K signaling pathway in malignant tumors. Therefore, this study investigated the expressions of molecules mentioned above and their relations in colorectal cancer cells.Methods In colorectal cancer HT29 and SW480 cells after treatment with Maraviroc (CCR5 inhibitor), MG132 (TRAF6 inhibitor), or NF-BAY (NF-κB inhibitor), the changes in the expressions of those proteins, as well as in proliferation, migration and invasion abilities were detected by Western blot, CCK-8 assay and Transwell assay, respectively.Results In the two types of colorectal cancer cells, the PI3K expression was decreased and the PTEN expression was increased (all P<0.05), while the expressions of TRAF6 and NF-κB showed no significant changes (all P>0.05) after inhibition of CCR5 protein; the expressions of PI3K and CCR5 were decreased and PTEN expression was increased (all P<0.05), while the NF-κB expression did not significantly change (both P>0.05) after inhibition of TRAF6 protein; the expressions of CCR5, TRAF6 and PI3K were decreased and PTEN expression was increased after inhibition of NF-κB expression (all P<0.05). The proliferation, migration and invasion abilities in the two types of colorectal cancer cells were significantly suppressed by the treatment of any of the three inhibitors (all P<0.05).Conclusion There is constitutive activation of the NF-κB in colorectal cancer cells, which may inhibit the activity of tumor suppressor molecule PTEN, and thereby lead to the increased activity of tumor-promoting molecule PI3K as well as its pathway through up-regulating the expressions of TRAF6 and CCR5.

    图1 CCR5抑制剂处理细胞后蛋白表达变化 1)与0 h比较,P<0.05Fig.1 Changes in protein expressions after treatment of CCR5 inhibitor 1) P<0.05 vs. 0-h value
    图2 TRAF6抑制剂处理细胞后蛋白表达变化 1)与0 h比较,P<0.05Fig.2 Changes in protein expressions after treatment of TRAF6 inhibitor 1) P<0.05 vs. 0-h value
    图3 NF-κB抑制剂处理细胞后蛋白表达变化 1)与0 h比较,P<0.05Fig.3 Changes in protein expressions after treatment of NF-κB inhibitor 1) P<0.05 vs. 0-h value
    图4 CCK-8分析细胞增殖Fig.4 Proliferation analysis by CCK-8 assay
    图5 Transwell分析细胞迁移和侵袭能力 1)与对照组比较,P<0.05Fig.5 Determination of cell migration and invasion abilities by Transwell assay 1) P<0.05 vs. control group
    图1 CCR5抑制剂处理细胞后蛋白表达变化 1)与0 h比较,P<0.05Fig.1 Changes in protein expressions after treatment of CCR5 inhibitor 1) P<0.05 vs. 0-h value
    图2 TRAF6抑制剂处理细胞后蛋白表达变化 1)与0 h比较,P<0.05Fig.2 Changes in protein expressions after treatment of TRAF6 inhibitor 1) P<0.05 vs. 0-h value
    图3 NF-κB抑制剂处理细胞后蛋白表达变化 1)与0 h比较,P<0.05Fig.3 Changes in protein expressions after treatment of NF-κB inhibitor 1) P<0.05 vs. 0-h value
    图4 CCK-8分析细胞增殖Fig.4 Proliferation analysis by CCK-8 assay
    图5 Transwell分析细胞迁移和侵袭能力 1)与对照组比较,P<0.05Fig.5 Determination of cell migration and invasion abilities by Transwell assay 1) P<0.05 vs. control group
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何亚琴,苏进达,赵少辉,王健,谢小亮.结直肠癌细胞中NF-κB活化与TRAF6、CCR5及PTEN/PI3K通路的关系及作用[J].中国普通外科杂志,2022,31(10):1363-1372.
DOI:10.7659/j. issn.1005-6947.2022.10.012

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  • 收稿日期:2022-08-16
  • 最后修改日期:2022-09-21
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  • 在线发布日期: 2022-10-31