Betulinic Acid Inhibits the Metastasis of Colon Cancer Cells through Regulating TGF-β/Smad Signaling Pathway

Objective:To explore the effect of betulinic acid (BA) on the growth and metastasis of colon cancer cell line HCT116 and the underlying possible mechanism.Methods:HCT116 cells were cultured with McCoy's 5A medium containing 10%fetal bovine serum and 1%penicillin/streptomycin mixed solution in a...

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Main Authors: Xiaoying LIN, Aling SHEN, Meizhu WU, Ying CHENG, Xiaoping CHEN, Huixin LIU, Liya LIU, Zhiqing SHEN, Xiangyan WU, Nanhui XU, Jianfeng CHU, Youqin CHEN, Jun PENG
Format: Article
Language:English
Published: Editorial Office of Rehabilitation Medicine 2020-02-01
Series:康复学报
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Online Access:http://kfxb.publish.founderss.cn/thesisDetails#10.3724/SP.J.1329.2020.01012
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author Xiaoying LIN
Aling SHEN
Meizhu WU
Ying CHENG
Xiaoping CHEN
Huixin LIU
Liya LIU
Zhiqing SHEN
Xiangyan WU
Nanhui XU
Jianfeng CHU
Youqin CHEN
Jun PENG
author_facet Xiaoying LIN
Aling SHEN
Meizhu WU
Ying CHENG
Xiaoping CHEN
Huixin LIU
Liya LIU
Zhiqing SHEN
Xiangyan WU
Nanhui XU
Jianfeng CHU
Youqin CHEN
Jun PENG
author_sort Xiaoying LIN
collection DOAJ
description Objective:To explore the effect of betulinic acid (BA) on the growth and metastasis of colon cancer cell line HCT116 and the underlying possible mechanism.Methods:HCT116 cells were cultured with McCoy's 5A medium containing 10%fetal bovine serum and 1%penicillin/streptomycin mixed solution in a 37℃,5%CO<sub>2</sub>incubator. HCT116 cells were seeded at a density of 0.2×10<sup>5</sup>cells/mL in a 96-well plate and cultured overnight. Then the cells were treated with different concentrations (0,10,20, and 40 μmol/L) of BA for 24,48 and 72 h, respectively. And then CCK8 was added to detect its absorbance (<italic>A</italic> value) at 450 nm. HCT116 cells were seeded at a density of 0.4×10<sup>5</sup>cells/mL in a 12-well plate and cultured overnight. The cells were then collected by digestion and centrifugation, and stained with trypan blue after treating with different concentrations (0,10,20, and 40 μmol/L) of BA for 48 h. The total number of cells was calculated by using a Countstar BioMed automatic cell counter. HCT116 cells were seeded at a density of 0.4×10<sup>5</sup>cells/mL in a 12-well plate and cultured overnight. Then the cells were collected by digestion and centrifugation after treating with different concentrations (0,10,20, and 40 μmol/L) of BA for 48 h.1.0×10<sup>5</sup>cells/mL cell suspension was used for the Transwell experiment of cell migration and invasion, respectively. The effects of different concentrations of BA intervention for 48 h on the migration and invasion ability of HCT116 were analyzed. Western blot was used to detect the effect of BA on the protein expression of TGF-β and Smad2/3 in HCT116 cells after treated with BA for 48 h.Results:(1) CCK8 assay showed that the cell viability in each group was significantly reduced after treatment with different concentrations of BA for 24,48 and 72 h, and the differences were statistically significant compared with the 0 μmol/L group respectively (<italic>P</italic>&lt;0.05); The cell count results showed the number of cells in each group was remarkably decreased, and the differences were statistically significant compared with the 0 μmol/L group respectively (<italic>P</italic>&lt;0.05). (2) Transwell assay showed that the number of cells that migrated or invaded into the membrane was profoundly inhibited in each group after treatment with different concentrations of BA for 48 h, and the differences were statistically significant compared with the 0 μmol/L group respectively (<italic>P</italic>&lt;0.05). (3) Western blot assay showed that the protein expression of TGF-β and Smad2/3 was obviously decreased in each group after treated with different concentrations of BA for 48 h compared with the 0 μmol/L group.Conclusion:BA can inhibit the growth and metastasis of colon cancer cells, and its mechanism may be achived by regulating the TGF-β/Smad signaling pathway.
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record_format Article
series 康复学报
spelling doaj-art-b0bcac21d24d46d9b078e902d18031d02025-01-14T10:04:08ZengEditorial Office of Rehabilitation Medicine康复学报2096-03282020-02-0130586323131203Betulinic Acid Inhibits the Metastasis of Colon Cancer Cells through Regulating TGF-β/Smad Signaling PathwayXiaoying LINAling SHENMeizhu WUYing CHENGXiaoping CHENHuixin LIULiya LIUZhiqing SHENXiangyan WUNanhui XUJianfeng CHUYouqin CHENJun PENGObjective:To explore the effect of betulinic acid (BA) on the growth and metastasis of colon cancer cell line HCT116 and the underlying possible mechanism.Methods:HCT116 cells were cultured with McCoy's 5A medium containing 10%fetal bovine serum and 1%penicillin/streptomycin mixed solution in a 37℃,5%CO<sub>2</sub>incubator. HCT116 cells were seeded at a density of 0.2×10<sup>5</sup>cells/mL in a 96-well plate and cultured overnight. Then the cells were treated with different concentrations (0,10,20, and 40 μmol/L) of BA for 24,48 and 72 h, respectively. And then CCK8 was added to detect its absorbance (<italic>A</italic> value) at 450 nm. HCT116 cells were seeded at a density of 0.4×10<sup>5</sup>cells/mL in a 12-well plate and cultured overnight. The cells were then collected by digestion and centrifugation, and stained with trypan blue after treating with different concentrations (0,10,20, and 40 μmol/L) of BA for 48 h. The total number of cells was calculated by using a Countstar BioMed automatic cell counter. HCT116 cells were seeded at a density of 0.4×10<sup>5</sup>cells/mL in a 12-well plate and cultured overnight. Then the cells were collected by digestion and centrifugation after treating with different concentrations (0,10,20, and 40 μmol/L) of BA for 48 h.1.0×10<sup>5</sup>cells/mL cell suspension was used for the Transwell experiment of cell migration and invasion, respectively. The effects of different concentrations of BA intervention for 48 h on the migration and invasion ability of HCT116 were analyzed. Western blot was used to detect the effect of BA on the protein expression of TGF-β and Smad2/3 in HCT116 cells after treated with BA for 48 h.Results:(1) CCK8 assay showed that the cell viability in each group was significantly reduced after treatment with different concentrations of BA for 24,48 and 72 h, and the differences were statistically significant compared with the 0 μmol/L group respectively (<italic>P</italic>&lt;0.05); The cell count results showed the number of cells in each group was remarkably decreased, and the differences were statistically significant compared with the 0 μmol/L group respectively (<italic>P</italic>&lt;0.05). (2) Transwell assay showed that the number of cells that migrated or invaded into the membrane was profoundly inhibited in each group after treatment with different concentrations of BA for 48 h, and the differences were statistically significant compared with the 0 μmol/L group respectively (<italic>P</italic>&lt;0.05). (3) Western blot assay showed that the protein expression of TGF-β and Smad2/3 was obviously decreased in each group after treated with different concentrations of BA for 48 h compared with the 0 μmol/L group.Conclusion:BA can inhibit the growth and metastasis of colon cancer cells, and its mechanism may be achived by regulating the TGF-β/Smad signaling pathway.http://kfxb.publish.founderss.cn/thesisDetails#10.3724/SP.J.1329.2020.01012colon cancerbetulinic acidtransferTGF-β/Smad signaling pathway
spellingShingle Xiaoying LIN
Aling SHEN
Meizhu WU
Ying CHENG
Xiaoping CHEN
Huixin LIU
Liya LIU
Zhiqing SHEN
Xiangyan WU
Nanhui XU
Jianfeng CHU
Youqin CHEN
Jun PENG
Betulinic Acid Inhibits the Metastasis of Colon Cancer Cells through Regulating TGF-β/Smad Signaling Pathway
康复学报
colon cancer
betulinic acid
transfer
TGF-β/Smad signaling pathway
title Betulinic Acid Inhibits the Metastasis of Colon Cancer Cells through Regulating TGF-β/Smad Signaling Pathway
title_full Betulinic Acid Inhibits the Metastasis of Colon Cancer Cells through Regulating TGF-β/Smad Signaling Pathway
title_fullStr Betulinic Acid Inhibits the Metastasis of Colon Cancer Cells through Regulating TGF-β/Smad Signaling Pathway
title_full_unstemmed Betulinic Acid Inhibits the Metastasis of Colon Cancer Cells through Regulating TGF-β/Smad Signaling Pathway
title_short Betulinic Acid Inhibits the Metastasis of Colon Cancer Cells through Regulating TGF-β/Smad Signaling Pathway
title_sort betulinic acid inhibits the metastasis of colon cancer cells through regulating tgf β smad signaling pathway
topic colon cancer
betulinic acid
transfer
TGF-β/Smad signaling pathway
url http://kfxb.publish.founderss.cn/thesisDetails#10.3724/SP.J.1329.2020.01012
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