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中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (4): 350-353.doi: 10.12047/j.cjap.5925.2020.075

• 研究论文 • 上一篇    下一篇

苦参素复合长春新碱对HCT-8/VCR细胞耐药性的影响及其机制*

苏建伟1, 周喜汉1△, 叶颖霞2, 蒋旗1   

  1. 1.右江民族医学院附属医院消化内科,
    2.右江民族医学院附属医院内科, 广西 百色 533000
  • 收稿日期:2019-08-13 修回日期:2020-05-27 发布日期:2020-11-09
  • 通讯作者: Tel: 13977698675; E-mail: 13977698675@163.com
  • 基金资助:
    *国家自然科学基金(81760739);2017年度广西高校中青年教师基础能力提升项目(2017KY0519);广西高校桂西地区高发病防治研究重点实验室开放课题(kfkt2016010)

Effects of oxymatrine and vincristine on drug resistance in HCT-8/VCR cells and its mechanism

SU Jian-wei1, ZHOU Xi-han1, YE Ying-xia2, JIANG Qi1   

  1. 1. Department of Gastroenterology,
    2. Department of Internal Medicine, Affiliated Hospital of Youjiang Medical College for nationalities, Baise 533000, China
  • Received:2019-08-13 Revised:2020-05-27 Published:2020-11-09

摘要: 目的: 探究苦参素复合长春新碱对HCT-8/VCR细胞耐药性的影响及其机制。方法: HCT-8/VCR体外培养,分为空白组、苦参素单药组,长春新碱单药组、苦参素和长春新碱联合组,每组6个复孔。CCK-8法检测各组HCT-8/VCR细胞的耐药性;MDC法检测细胞内自噬小泡的变化;ELISA法检测IL-6的含量;Western blot检测自噬相关基因与免疫因子TLR4蛋白的表达水平。结果: 苦参素复合长春新碱可降低HCT-8/VCR细胞的耐药性,逆转倍数为3.23;与空白组比较,苦参素组和联合组自噬体含量降低、IL-6含量也明显降低(P<0.01),长春新碱组自噬体含量升高,IL-6含量也明显升高(P<0.01);联合组与苦参素组比较,自噬体含量升高,而IL-6含量降低(P< 0.01);Western blot实验表明,与空白组比较,苦参素组P62的表达降低(P<0.05),LC3-Ⅱ/LC3-Ⅰ、Beclin-1、TLR4均升高(P<0.05),长春新碱组和联合组P62的表达升高(P<0.05),LC3-Ⅱ/LC3-Ⅰ、Beclin-1、TLR4均降低(P< 0.05);联合组与长春新碱组比较,P62的表达升高(P<0.05),LC3-Ⅱ/LC3-Ⅰ、Beclin-1、TLR4均降低(P<0.05)。结论: 苦参素与长春新碱联合可显著降低HCT-8/VCR的耐药性,其机制与抑制自噬活动和TLR4信号活化有关。

关键词: 苦参素, HCT-8/VCR细胞, 自噬基因, TLR4, 结肠癌, 长春新碱, 耐药

Abstract: Objective: To study the effects of oxymatrine and vincristine on resistance in HCT-8/VCR cells and its mechanism. Methods: HCT-8 / VCR cells were cultured in vitro and were divided into blank control group, oxymatrine group, vincristine group, oxymatrine and vincristine combined group, each group had 6 complexes. The drug resistance of HCT-8/VCR cells was investigated by CCK-8 when treated with vincristine alone or in combination with oxymatrine. The autophagy was determined by monodansylcadaverine (MDC) staining. The level of IL-6 was detected by ELISA. The expressions of autophagy-related gene P62, LC3-Ⅱ / LC3-Ⅰ, Beclin-1 protein and TLR4 were detected by Western blot assay. Results: Oxymatrine combined with vincristine could reduce the drug resistance of HCT-8 / VCR cells by the reversal multiple of 3.23. Compared with the blank control group, the content of autophagosome and the content of IL-6 in the oxymatrine group and the combination group were also decreased significantly (P<0.01). The content of autophagosome in the vincristine group was increased and the content of IL-6 was also significantly increased (P<0.01). Compared with the oxymatrine group, the combination group had higher autophagosome content, while IL-6 content was decreased (P<0.01); Western blot experiments showed that compared with the blank control group, the expression of P62 in the oxymatrine group was decreased (P<0.05), while the expressions of LC3-Ⅱ / LC3-Ⅰ, Beclin-1 and TLR4 were all increased (P<0.05). The expression of P62 in the vincristine group and the combined group was increased (P<0.05), and the expressions of LC3-Ⅱ / LC3-Ⅰ, Beclin-1, and TLR4 were all decreased (P<0.05). Compared with the vincristine group, the expression of P62 was increased in the combination group (P<0.05), and the expressions of LC3-Ⅱ / LC3-Ⅰ, Beclin-1, and TLR4 were all decreased (P<0.05). Conclusion: Oxymatrine combined with vincristine can reduce the drug resistance of HCT-8/VCR cells, which may be related to the regulation of autophagy activity and TLR4 signal activation.

Key words: oxymatrine, HCT-8/VCR cells, autophagy, TLR4, colorectal cancer, vincristine, reversal

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