黄芪-莪术-重楼配伍激活PERK/eIF2α/ATF4信号通路介导抑制结直肠癌生长转移作用研究

Astragalus Mongholicus Bunge-Curcuma Aromatica-Paridis Rhizoma Inhibits Metastasis of Colon Cancer via the PERK/eIF2α/ATF4 Signaling Pathway

    摘要
  • 摘要:
      目的  探讨黄芪-莪术-重楼(芪-术-蚤)配伍通过激活PERK/eIF2α/ATF4信号通路介导内质网应激(ER stress)抗结直肠癌生长、转移相关作用。
      方法  BALB/c雄性小鼠30只随机分为假手术组、模型组、5-氟尿嘧啶(5-FU)组(25 mg·kg-1)、芪-术-蚤高剂量组(5.85 g·kg-1)、芪-术-蚤低剂量组(2.925 g·kg-1)，每组6只, 构建结直肠癌原位移植瘤小鼠模型, 给药15 d后, 通过肿瘤体积大小变化评估芪-术-蚤配伍对肿瘤生长的干预效果; 通过HE染色评估芪-术-蚤角药配伍干预后小鼠肝脏组织、肿瘤组织变化情况。采用ELISA法检测丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)水平变化; Western blot检测PERK/eIF2α/ATF4信号通路和上皮细胞-间充质转化(EMT)相关蛋白表达。
      结果  与模型组相比, 芪-术-蚤高剂量组肿瘤体积明显减小(P＜0.000 1)、肝脾转移灶较不明显, 肝组织和肿瘤组织病理染色结果产生变化, 下调氧化应激相关指标GSH-Px、SOD水平, 上调MDA水平, 上调ER stress相关蛋白磷酸化蛋白激酶R样内质网激酶(p-PERK)、磷酸化真核启动因子2α(p-eIF2α)、转录激活因子4(ATF4)等的表达, 上调上皮型钙黏蛋白(E-cadherin)、下调神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、锌指转录因子(Snail)的蛋白表达水平, 抑制EMT过程。
      结论  芪-术-蚤角药配伍可能通过激活PERK/eIF2α/ATF4通路诱导持续的ER stress, 影响结直肠癌EMT过程, 从而抑制结直肠癌的生长转移。

     

    Abstract:
      OBJECTIVE  To investigate the mechanism of anti-colorectal cancer growth and metastasis-related effects of Astragalus mongholicus Bunge-Curcuma aromatica-Paridis Rhizoma (Qi-Zhu-Zao) pairing through PERK/eIF2α/ATF4 signaling pathway mediating endoplasmic reticulum stress.
      METHODS  Twenty-four BALB/c male mice were randomly divided into sham-operated group, model group, 5-FU (5-fluorouracil) group (25 mg·kg-1), and Qi-Zhu-Zao high dose group (5.85 g·kg-1), Qi-Zhu-Zao low dose group(2.925 g·kg-1) (n=6) to construct a mouse model of colorectal cancer in situ transplantation tumor, and the intervention effect of Qi-Zhu-Zao combination on tumor growth was assessed by the change of tumor volume size after 15 days of administration; the intervention effect of Qi-Zhu-Zao combination on tumor growth was assessed by H&E. Pathological staining was used to evaluate the effect of Qi-Zhu-Zao combination on the liver and tumor tissues of mice. The changes of MDA, SOD and GSH-Px levels were detected by enzyme-linked immunosorbent assay (ELISA); the expression of PERK/eIF2α/ATF4 signaling pathway and EMT-related proteins were detected by protein immunoblotting (Western blot).
      RESULTS  Compared with the model group, the tumor volume was significantly reduced (P < 0.000 1), liver and spleen metastases were less pronounced in the Qi-Zhu-Zao high-dose group, and histopathological staining results of liver tissue and tumor produced changes in oxidative stress indicators SOD, MDA, and GSH-Px, upregulation of ER stress-related proteins p-PERK, p-IF2α, and ATF4, etc., upregulated the protein expression levels of E-Cadherin, downregulated N-Cadherin, Vimentin, and Snail, and inhibited the EMT process (P < 0.01 or P < 0.05).
      CONCLUSION  In this paper, we investigated the regulatory mechanism related to the inhibition of colorectal cancer growth and metastasis by the combination of Qi-Zhu-Zao trigonal medicine, and demonstrated that it may inhibit the growth and metastasis of colorectal cancer by activating the PERK/eIF2α/ATF4 pathway to induce sustained ER stress and affect the EMT process of colorectal cancer.

     

    • HTML全文
    • 参考文献(24)
    • 相关文章
    • 施引文献
  • 资源附件(0)

/

返回文章
返回