Anti-colon cancer activity and effect on autophagy of active fraction from clove in vitro

WANG Jian; LIU Lin; YANG Xiaohui; ZHANG Hong; ZHAO Ge; LIU Minghua

doi:10.3969/j.issn.2096-3351.2023.02.016

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Anti-colon cancer activity and effect on autophagy of active fraction from clove in vitro

更新时间：2023-05-05

- Anti-colon cancer activity and effect on autophagy of active fraction from clove in vitro
- Journal of Southwest Medical University Vol. 46, Issue 2, Pages: 167-170(2023)
- 作者机构：
  
  1. 西南医科大学药学院药理教研室,泸州,646000
  2. 西南医科大学附属医院药学部,泸州,646000
- 作者简介：
- 基金信息：
- DOI：10.3969/j.issn.2096-3351.2023.02.016
  CLC：
- Published：2023，
  
  Published Online：20 March 2023，
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WANG Jian, LIU Lin, YANG Xiaohui, et al. Anti-colon cancer activity and effect on autophagy of active fraction from clove in vitro. [J]. Journal of Southwest Medical University 46(2):167-170(2023)
DOI：

WANG Jian, LIU Lin, YANG Xiaohui, et al. Anti-colon cancer activity and effect on autophagy of active fraction from clove in vitro. [J]. Journal of Southwest Medical University 46(2):167-170(2023) DOI： 10.3969/j.issn.2096-3351.2023.02.016.

摘要

目的体外实验评价丁香活性组分（active fraction from clove，AFC）的抗结肠癌活性，探讨AFC 对敏感结肠癌细胞自噬的影响。方法采用Cell Counting Kit-8 assay 检测AFC 对多株结肠癌细胞LOVO、HCT116、HCT8、SW620 增殖的抑制作用，体外筛选敏感的人结肠癌细胞。GFP-LC3 转染后荧光显微镜检测自噬水平的强弱，透射电镜观察自噬体和自噬溶酶体的形成，Western Blot 检测自噬相关蛋白LC3、Beclin-1 的表达。结果AFC 对LOVO、HCT116、HCT8、SW620 细胞的IC50分别是280.90 ± 12.61、113.50 ± 7.83、211.71 ± 7.29、174.92 ± 9.52μg/mL，最敏感的细胞是HCT116，最佳干预时间为48 h。AFC 处理细胞48 h 后，荧光显微镜观察到HCT116 细胞GFP-LC3 转染后出现明显自噬斑点，透射电镜观察到HCT116 细胞大量的自噬体和自噬溶酶体形成。Western blot 结果表明，HCT116 细胞AFC 处理组自噬相关蛋白LC3Ⅱ/LC3Ⅰ、Beclin-1 相对表达量明显增高，呈现一定的时间与浓度依赖性，与对照组细胞比较差异明显（P < 0.05）。结论AFC 抗结肠癌活性较强，对HCT116 细胞最敏感，可能与其上调自噬相关蛋白LC3Ⅱ/LC3Ⅰ、Beclin-1 的表达，诱导HCT116 细胞发生自噬有关。

Abstract

Objective To study the anti-colon cancer activity of active fraction from clove（AFC) and the effect of AFC on autophagy of sensitive colon cancer cells in vitro. Methods Cell Counting Kit-8 assay was used to detect the inhibitory effect of AFC on the proliferation of Human Colon Cancer Cells LOVO、HCT116、HCT8 and SW620，in order to screen the sensitive cells. The level of autophagy was detected by GFP-LC3 transfection，the formation of autophagy and autophagy lysosome were observed by transmission electron microscope，and the expression of LC3 and Beclin-1 were detected by western blotting. Results The IC50 values of Lovo cells， HCT116 cells，HCT8 cells and SW620 cells were 280.90 ± 12.61，113.50 ± 7.83，211.71 ± 7.29，174.92 ± 9.52 μg/mL，respectively. The most sensitive cells were HCT116 cells，and the optimal intervention time was 48 hours. After transfection with GFP-LC3，autophagy spots were observed in the HCT116 cell of AFC treatment by fluorescence microscope. A large number of autophagosomes and autophagolysosomes were observed by transmission electron microscope. Western blot showed that the relative expression levels of autophagy-related proteins LC3II/LC3I and Beclin-1 in the HCT116 cell of AFC treatment were increased in a concentrationdependent manner and time-dependent manner，compared with the control group cell，P < 0.05. Conclusion AFC showed strong anticolon cancer activity and it was most sensitive to HCT116 cells，which might be related to up-regulating autophagy-related protein LC3 ii/LC3 I，Beclin-1 expression and inducing autophagy in HCT116 cells.

关键词

丁香活性组分HCT116 细胞增殖自噬

Keywords

Active fraction from cloveHCT116 cellsProliferationAutophagy

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