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中华肝脏外科手术学电子杂志 ›› 2018, Vol. 07 ›› Issue (05) : 420 -425. doi: 10.3877/cma.j.issn.2095-3232.2018.05.017

所属专题: 文献

基础研究

青蒿素抑制胆囊癌细胞的迁移侵袭及其机制
秦一雨1, 靳龙洋2, 葛安兴3, 黑振宇2,()   
  1. 1. 224005 江苏省盐城市,江苏医药职业学院博士后创新实践基地;200092 上海交通大学医学院附属新华医院普通外科
    2. 200092 上海交通大学医学院附属新华医院普通外科
    3. 224005 江苏省盐城市,江苏医药职业学院博士后创新实践基地
  • 收稿日期:2018-07-04 出版日期:2018-10-10
  • 通信作者: 黑振宇
  • 基金资助:
    国家自然科学青年基金(81702422); 江苏省高等学校自然科学研究面上项目(17KJB320024); 江苏省卫生和计划生育委员会面上项目(H2017086); 江苏省卫生职业技术教育研究室项目(J201606); 盐城市科学技术局项目(YK2016044)

Effect and mechanism of artemisinin on inhibiting metastasis and invasion of gallbladder carcinoma cells

Yiyu Qin1, Longyang Jin2, Anxing Ge3, Zhenyu Hei2,()   

  1. 1. Postdoctoral Innovation Practice Base of Jiangsu Vocational College of Medicine, Yancheng 224005, China; Department of General Surgery, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
    2. Department of General Surgery, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
    3. Postdoctoral Innovation Practice Base of Jiangsu Vocational College of Medicine, Yancheng 224005, China
  • Received:2018-07-04 Published:2018-10-10
  • Corresponding author: Zhenyu Hei
  • About author:
    Corresponding author: Hei Zhenyu, Email:
引用本文:

秦一雨, 靳龙洋, 葛安兴, 黑振宇. 青蒿素抑制胆囊癌细胞的迁移侵袭及其机制[J]. 中华肝脏外科手术学电子杂志, 2018, 07(05): 420-425.

Yiyu Qin, Longyang Jin, Anxing Ge, Zhenyu Hei. Effect and mechanism of artemisinin on inhibiting metastasis and invasion of gallbladder carcinoma cells[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2018, 07(05): 420-425.

目的

探讨青蒿素对胆囊癌细胞体外迁移、侵袭能力的影响及其机制。

方法

20 μmol/L青蒿素处理胆囊癌细胞GBC-SD(青蒿素组),同时设立未加青蒿素处理的对照组。采用划痕实验检测胆囊癌细胞迁移能力变化,Transwell实验检测细胞侵袭能力变化,免疫荧光技术检测细胞形态变化。采用qRT-PCR和Western blot检测细胞中E-cadherin、Slug、Vimentin、MMP-2、MMP-9等基因的mRNA和蛋白水平变化。两组表达量比较采用t检验。

结果

划痕24 h青蒿素组划痕宽(393±23)μm,明显大于对照组的(210±15) μm (t=13.415,P<0.05)。青蒿素组GBC-SD穿膜细胞数为(93±8)个,明显少于对照组的(202±14)个(t=-17.037,P<0.05)。青蒿素促使细胞形态从间质型向表皮型转化。青蒿素组GBC-SD细胞的Slug、Vimentin、MMP-2、MMP-9基因的mRNA分别为0.89±0.03、1.35±0.10、3.30±0.13、1.15±0.08,明显低于对照组的2.08±0.12、3.22±0.15、4.72±0.19、1.95±0.15(t=-19.812,-20.775,-12.259,-9.438;P<0.05);而E-cadherin mRNA为2.06±0.08,明显高于对照组的1.03±0.06 (t=20.956;P<0.05)。Western blot检测显示青蒿素下调Slug、Vimentin、MMP-2、MMP-9,上调E-cadherin蛋白。

结论

青蒿素可抑制胆囊癌细胞迁移和侵袭,其作用可能与抑制Slug基因表达逆转上皮间质转化,进而降低MMP表达有关。

Objective

To investigate the effect of artemisinin on the in vitro metastasis and invasion of gallbladder carcinoma cells and its mechanism.

Methods

Gallbladder carcinoma cells GBC-SD was treated with 20 μmol/L artemisinin (artemisinin group) and cells without artemisinin treatment was established as control group. The metastasis ability of gallbladder carcinoma cell was detected using scratch wound assay. The variation of cell invasion was evaluated using Transwell assay. The change of cell morphology was observed using immunofluorescent staining. The mRNA and protein level changes of E-cadherin, Slug, Vimentin, MMP-2 and MMP-9 were detected by qRT-PCR and Western blot. The expression levels between two groups were compared by t test.

Results

At 24 h, the scratch width was (393±23) μm in artemisinin group, significantly wider than (210±15) μm in control group (t=13.415, P<0.05). In artemisinin group, the quantity of transmembrane GBC-SD cells was 93±8, significantly less than 202±14 in control group (t=-17.037, P<0.05). Artemisinin promoted the epithelial-mesenchymal transition of cell morphology. In artemisinin group, the expression levels of Slug, Vimentin, MMP-2 and MMP-9 mRNA of GBC-SD cells were 0.89±0.03, 1.35±0.10, 3.30±0.13 and 1.15±0.08, significantly lower than 2.08±0.12, 3.22±0.15, 4.72±0.19 and 1.95±0.15 in control group (t=-19.812, -20.775, -12.259, -9.438; P<0.05). In artemisinin group, the expression of E-cadherin mRNA was 2.06±0.08, significantly higher than 1.03±0.06 in control group (t=20.956; P<0.05). Western blot showed that artemisinin could down-regulate the expression levels of Slug, Vimentin, MMP-2 and MMP-9 proteins, whereas up-regulate the expression of E-cadherin protein.

Conclusions

Artemisinin can inhibit the metastasis and invasion of gallbladder carcinoma cells. It is probably related with suppressing the expression of Slug gene, reversing the epithelial-mesenchymal transition, thereby down-regulating the expression level of MMP.

表1 引物序列
图1 划痕实验检测青蒿素对GBC-SD细胞迁移的影响
图2 Transwell实验检测青蒿素对GBC-SD细胞侵袭的影响
图3 免疫荧光实验检测青蒿素对GBC-SD细胞形态的影响
图4 青蒿素对GBC-SD细胞内Slug、E-cadherin等基因蛋白水平的影响
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