Anticancer Effect of Isoliquiritigenin on Human Clear Cell Renal Cell Carcinoma 786-O Cells and Molecular Mechanism

Hong XIN; Peng-ran SUN; Peng SONG; Wei XU

doi:10.13422/j.cnki.syfjx.20191122

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Anticancer Effect of Isoliquiritigenin on Human Clear Cell Renal Cell Carcinoma 786-O Cells and Molecular Mechanism

Pharmacology | 更新时间：2021-02-09

- Anticancer Effect of Isoliquiritigenin on Human Clear Cell Renal Cell Carcinoma 786-O Cells and Molecular Mechanism
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 12, Pages: 83-89(2019)
- 作者机构：
  
  哈尔滨医科大学　附属第一医院，哈尔滨　 150001
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191122
  CLC： R22; R242; R2-031; R285.5
- Received：19 October 2018，
  
  Published Online：19 February 2019，
  
  Published：20 June 2019
- 稿件说明：
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Hong XIN, Peng-ran SUN, Peng SONG, et al. Anticancer Effect of Isoliquiritigenin on Human Clear Cell Renal Cell Carcinoma 786-O Cells and Molecular Mechanism[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(12): 83-89.
DOI：

Hong XIN, Peng-ran SUN, Peng SONG, et al. Anticancer Effect of Isoliquiritigenin on Human Clear Cell Renal Cell Carcinoma 786-O Cells and Molecular Mechanism[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(12): 83-89. DOI： 10.13422/j.cnki.syfjx.20191122.

摘要

目的：

研究异甘草素(isoliquiritigenin

ISL)对人肾透明细胞癌786-O细胞的抗癌作用，并探讨其可能存在的分子机制。

方法：

通过噻唑蓝(thiazolyl blue tetrazolium bromide，MTT)比色法检测ISL(0

25，50

100 μmol·L

－1

)对786-O细胞增殖的作用；细胞划痕和Transwell实验检测ISL对786-O细胞迁移、侵袭能力的影响；吖啶橙染色，Ad-GFP-LC3转染实验观察细胞自噬状态；蛋白免疫印迹法(Western blot)检测自噬相关蛋白的表达，并分析磷脂酰肌醇-3-激酶(phosphatidylinositol-3-kinase

PI3K)/蛋白激酶B(protein kinase B

Akt)/哺乳动物雷帕霉素蛋白(mammalian target of rapamycin

mTOR)信号通路变化，探讨其可能的作用机制。

结果：

MTT比色法结果显示，与空白组比较，ISL能够明显抑制786-O细胞的增殖，且呈时间-剂量依赖性(

0.05)；划痕和Transwell实验结果表明，与空白组比较，ISL能够抑制785-O细胞的迁移和侵袭(

0.05)；吖啶橙染色和Ad-GFP-LC3转染实验表明ISL能够诱导786-O细胞发生自噬。此外，与空白组比较，ISL能够诱导细胞内自噬标志物-Ⅱ(LC3-Ⅱ)，自噬相关蛋白Beclin1，Atg5蛋白表达(

0.05)，并显著下调p62蛋白表达(

0.05)，当加入自噬抑制剂3-甲基腺嘌呤(3-methyladenine

3-MA)可逆转上述现象(

0.05)；同时ISL能够抑制Akt，mTOR的磷酸化水平(

0.05)，加入抑制剂LY294002和雷帕霉素(rapamycin

Rap)，LC3-Ⅱ，Beclin1，Atg5蛋白表达明显上调(

0.05)，p62蛋白表达明显下调(

0.05)。

结论：

ISL能够抑制肾透明细胞癌786-O细胞的增殖、迁移和侵袭，并通过抑制PI3K/Akt/mTOR信号通路诱导细胞发生自噬。

Abstract

Objective:

To investigate the anticancer effect of isoliquiritigenin (ISL) on human clear cell renal cell carcinoma 786-O cells

and explore its possible molecular mechanism.

Method:

Thiazolyl blue tetrazolium bromide (MTT) assay was used to detect effect of ISL (0

100 μmol·L

-1

) on proliferation of 786-O cells. The effect of ISL on migration and invasion of 786-O cells was detected by cell scratch test and Transwell assay. The autophagy was observed under the fluorescence microscope through acridine orange staining and Ad-GFP-LC3 transfection experiment. Western blot was used to detect the expression of autophagy related protein and analyze the changes of phosphatidylinositol-3-kinase (PI3K)/protein kinase B(Akt)/mammalian target of rapamycin (mTOR) signaling pathway to explore the possible mechanism.

Result:

MTT results showed that ISL could significantly inhibit the proliferation of 786-O cells in a time-dose dependent manner (

0.05). The results of scratch and Transwell experiments indicated that ISL could inhibit the migration and invasion of 786-O cells (

0.05). Acridine orange staining and ad-GFP-LC3 transfection showed that ISL can induce autophagy in 786-O cells. Besides

ISL can induce LC3-Ⅱ

Beclin1

Atg5 protein expressions (

0.05)

and significantly reduce p62 protein expression (

0.05); autophagy inhibitor 3-methyl adenine (3-MA) can be added to reverse the above phenomena (

0.05). Meanwhile

ISL can inhibit phosphorylation of Akt

mTOR levels (

0.05)

inhibitor LY294002 and rapamycin (Rap) can be added to significantly re-regulate LC3-Ⅱ

Beclin1

Atg5 protein expressions (

0.05) and significantly down-regulate p62 protein expression (

0.05).

Conclusion:

ISL can inhibit the proliferation

migration and invasion of clear cell renal carcinoma 786-O cells

and induce autophagy by inhibiting the PI3K/Akt/mTOR signaling pathway.

关键词

Keywords

references

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