纳米化牛樟芝对人肝癌HepG2细胞迁移侵袭能力的影响
|
摘要
目的比较牛樟芝乙醇提取物(以下简称牛樟芝醇提物)及纳米化牛樟芝子实体(以下简称纳米牛樟芝)对肝癌细胞HepG2细胞迁移与侵袭能力的影响。方法以肝癌细胞HepG2为研究对象,分为牛樟芝醇提物组和纳米牛樟芝组,通过细胞划痕试验观察200μg·mL-1的浓度作用下,牛樟芝乙醇提取物与纳米化牛樟芝子实体对HepG2细胞迁移能力的影响;采用Transwell侵袭试验检测牛樟芝乙醇提取物与纳米化牛樟芝子实体对HepG2细胞侵袭能力的影响;最后采用蛋白免疫印迹检测E-cadherin、VEGF等细胞迁移侵袭相关蛋白的变化。结果 (1)细胞划痕试验结果显示,牛樟芝醇提物与纳米牛樟芝处理后,HepG2细胞迁移能力显著下降,且在划痕后24小时后,纳米化牛樟芝子实体组细胞迁移率低于牛樟芝乙醇提取物组,两者差异具有统计学意义(p<0.05)。(2)Transwell侵袭试验表明, 200μg·mL-1牛樟芝醇提物与纳米牛樟芝作用HepG2细胞24小时后穿过Matrigel胶数显著减少(p <0.001),且纳米化牛樟芝子实体组细胞数低于牛樟芝醇提物组。(3)牛樟芝醇提物与纳米牛樟芝可上调E-cadherin的表达,但对VEGF的表达无明显调节作用。结论牛樟芝乙醇提取物与纳米化牛樟芝子实体均有较好抑制肝癌细胞迁移侵袭的活性,且纳米化牛樟芝子实体抑制细胞迁移侵袭活性优于牛樟芝乙醇提取物。
Objective: To compare the effect of antrodia camphorata extract and Nano-antrodia camphorata fruiting body on migration and invasion of hepatocellular carcinoma HepG2 cells. Methods: The hepatocellular carcinoma HepG2 cells were selected as the objects of study divided into extract group of antrodia camphorate, and fruiting body group of Nano-antrodia camphorate. In two groups, the migration of HepG2 cells were observed by Cell Scratch Test, and the invasion of HepG2 cells were detected by Transwell Assay. In addition, the associated protein of HepG2 in E-cadherin and VEG were detected by Western Blotting. Results:(1) After 24 hours, the Cell Scratch Test showed that two groups were significant decreased, but fruiting body group of Nano-antrodia camphorate decreased more obviously(P<0.05).(2)After 24 hours, the Transwell Assay showed that the number of Matrigel in two groups had significantly decreased, but fruiting body group of Nano-antrodia camphorate was more obviously(p<0.001).(3) The expression of E-cadherin was increased in two groups. However, there was no significant effect on the expression of VEGF. Conclusions: Antrodia camphorate has significant effects on migration and invasion of hepatocellular carcinoma HepG2 cells. Moreover, Nano-antrodia camphorata fruiting body is superior to antrodia camphorata extract.
Objective: To compare the effect of antrodia camphorata extract and Nano-antrodia camphorata fruiting body on migration and invasion of hepatocellular carcinoma HepG2 cells. Methods: The hepatocellular carcinoma HepG2 cells were selected as the objects of study divided into extract group of antrodia camphorate, and fruiting body group of Nano-antrodia camphorate. In two groups, the migration of HepG2 cells were observed by Cell Scratch Test, and the invasion of HepG2 cells were detected by Transwell Assay. In addition, the associated protein of HepG2 in E-cadherin and VEG were detected by Western Blotting. Results:(1) After 24 hours, the Cell Scratch Test showed that two groups were significant decreased, but fruiting body group of Nano-antrodia camphorate decreased more obviously(P<0.05).(2)After 24 hours, the Transwell Assay showed that the number of Matrigel in two groups had significantly decreased, but fruiting body group of Nano-antrodia camphorate was more obviously(p<0.001).(3) The expression of E-cadherin was increased in two groups. However, there was no significant effect on the expression of VEGF. Conclusions: Antrodia camphorate has significant effects on migration and invasion of hepatocellular carcinoma HepG2 cells. Moreover, Nano-antrodia camphorata fruiting body is superior to antrodia camphorata extract.
引文
[1]Chang T T,Chou W N.Antrodia cinnamomea sp.nov.on Cinnamomum kanehirai in Taiwan.[J].Mycological Research,1995,99(6):756-758.
[2]张远腾,李晓波.牛樟芝化学成分及其药理作用研究进展[J].中草药,2016(06):1034-1042.
[3]Fitzmaurice C,Akinyemiju T F,Al L F,et al.Global,Regional,and National Cancer Incidence,Mortality,Years of Life Lost,Years Lived With Disability,and Disability-Adjusted Life-Years for 29 Cancer Groups,1990 to2016:A Systematic Analysis for the Global Burden of Disease Study[J].JAMAOncol,2018.
[4]李红玉,郭茂田,薛博瑜.牛樟芝提取物对人肝癌SMMC-7721细胞增殖的影响[J].中国实验方剂学杂志,2015(10):118-122.
[5]Hseu Y C,Chang G R,Pan J Y,et al.Antrodia camphorata inhibits epithelial-to-mesenchymal transition by targeting multiple pathways in triple-negative breast cancers[J].J Cell Physiol,2018.
[6]Wang G,Wan Y,Zhao J,et al.Ethanol extract of Antrodia camphorata inhibits proliferation of HCT-8 human colorectal cancer cells by arresting cell cycle progression and inducing apoptosis[J].Mol Med Rep,2017,16(4):4941-4947.
[7]Li Z W,Kuang Y,Tang S N,et al.Hepatoprotective activities of Antrodia camphorata and its triterpenoid compounds against CCl4-induced liver injury in mice[J].J Ethnopharmacol,2017,206:31-39.
[8]Wang J J,Wu C C,Lee C L,et al.Antimelanogenic,Antioxidant and Antiproliferative Effects of Antrodia camphorata Fruiting Bodies on B16-F0Melanoma Cells[J].PLoS One,2017,12(1):e170924.
[9]Chang J S,Kuo H P,Chang K L,et al.Apoptosis of Hepatocellular Carcinoma Cells Induced by Nanoencapsulated Polysaccharides Extracted from Antrodia Camphorata[J].PLoS One,2015,10(9):e136782.
[10]Lee C I,Wu C C,Hsieh S L,et al.Anticancer effects on human pancreatic cancer cells of triterpenoids,polysaccharides and 1,3-beta-D-glucan derived from the fruiting body of Antrodia camphorata[J].Food Funct,2014,5(12):3224-3232.
[11]Kuo P L,Hsu Y L,Cho C Y,et al.Apoptotic effects of Antrodia cinnamomea fruiting bodies extract are mediated through calcium and calpain-dependent pathways in Hep 3B cells[J].Food Chem Toxicol,2006,44(8):1316-1326.
[2]张远腾,李晓波.牛樟芝化学成分及其药理作用研究进展[J].中草药,2016(06):1034-1042.
[3]Fitzmaurice C,Akinyemiju T F,Al L F,et al.Global,Regional,and National Cancer Incidence,Mortality,Years of Life Lost,Years Lived With Disability,and Disability-Adjusted Life-Years for 29 Cancer Groups,1990 to2016:A Systematic Analysis for the Global Burden of Disease Study[J].JAMAOncol,2018.
[4]李红玉,郭茂田,薛博瑜.牛樟芝提取物对人肝癌SMMC-7721细胞增殖的影响[J].中国实验方剂学杂志,2015(10):118-122.
[5]Hseu Y C,Chang G R,Pan J Y,et al.Antrodia camphorata inhibits epithelial-to-mesenchymal transition by targeting multiple pathways in triple-negative breast cancers[J].J Cell Physiol,2018.
[6]Wang G,Wan Y,Zhao J,et al.Ethanol extract of Antrodia camphorata inhibits proliferation of HCT-8 human colorectal cancer cells by arresting cell cycle progression and inducing apoptosis[J].Mol Med Rep,2017,16(4):4941-4947.
[7]Li Z W,Kuang Y,Tang S N,et al.Hepatoprotective activities of Antrodia camphorata and its triterpenoid compounds against CCl4-induced liver injury in mice[J].J Ethnopharmacol,2017,206:31-39.
[8]Wang J J,Wu C C,Lee C L,et al.Antimelanogenic,Antioxidant and Antiproliferative Effects of Antrodia camphorata Fruiting Bodies on B16-F0Melanoma Cells[J].PLoS One,2017,12(1):e170924.
[9]Chang J S,Kuo H P,Chang K L,et al.Apoptosis of Hepatocellular Carcinoma Cells Induced by Nanoencapsulated Polysaccharides Extracted from Antrodia Camphorata[J].PLoS One,2015,10(9):e136782.
[10]Lee C I,Wu C C,Hsieh S L,et al.Anticancer effects on human pancreatic cancer cells of triterpenoids,polysaccharides and 1,3-beta-D-glucan derived from the fruiting body of Antrodia camphorata[J].Food Funct,2014,5(12):3224-3232.
[11]Kuo P L,Hsu Y L,Cho C Y,et al.Apoptotic effects of Antrodia cinnamomea fruiting bodies extract are mediated through calcium and calpain-dependent pathways in Hep 3B cells[J].Food Chem Toxicol,2006,44(8):1316-1326.