肝癌HepG2细胞分泌的Exosome对间充质干细胞分化的影响及其相互作用机制
Effect of Human Hepatocellular Carcinoma HepG2 Cell-derived Exosome on the Differentiation of Mesenchymal Stem Cells and Their Interaction
查看参考文献15篇
|
文摘
|
目的评估肝癌HepG2细胞分泌的Exosome对间充质干细胞(MSC)分化为肿瘤相关成纤维细胞(CAF)的影响及其相互作用机制。方法在人脂肪MSC培养体系中加入HepG2细胞分泌的Exosome进行培养,采用Western blot法检测CAF特征性蛋白的表达情况。将已鉴定为CAF样条件培养基(CAF-CM)和牛血清白蛋白对照组分别加至HepG2培养基中,采用Western blot法和定量PCR检测上皮间质转化相关基因的表达,MTS法检测细胞增殖,Transwell法检测HepG2细胞的迁移和侵袭。结果HepG2细胞分泌的Exosome表达CD63、热休克蛋白(HSP)70和HSP90。将Exosome加入MSC培养体系后14 d,可检测到α平滑肌肌动蛋白、成纤维细胞激活蛋白α、白细胞介素(IL)-6、IL-8和IL-1β等CAF特征性蛋白的表达。MTS法检测CAF-CM组的细胞增殖OD值为1.075±0.104,明显高于对照组的0.874±0.066(P=0.023)和MSC条件培养基组的0.649±0.034(P=0.0005)。CAF-CM组迁移的HepG2细胞数为(42.5±9.1)个,明显高于对照组的(18.5±3.1)个(P=0.001);CAF-CM组侵袭细胞数为(29.0±3.5)个,明显高于对照组的(13.1±3.7)个(P=0.009)。CAF-CM组Smad交互蛋白1(P=0.040)、β-catenin(P=0.038)、纤连蛋白(P=0.029)和Vimentin(P=0.013)的表达较对照组显著上调;而CAF-CM组的紧密连接蛋白表达显著低于对照组(P=0.010)。结论肝癌HepG2细胞系分泌的Exosome可诱导脂肪组织来源的MSC分化为CAF,后者又可以促进HepG2细胞的增殖、迁移和侵袭。 |
|
其他语种文摘
|
To investigate the effect of human hepatocellular carcinoma HepG2 cell-derived Exosome on the differentiation of mesenchymal stem cells(MSC)into cancer-associated myofibroblasts(CAF)and the impacts of CAF on liver cancer cell proliferation,migration,and invasion. Methods The protein expression of HepG2 cell-derived Exosome was detected by Western blotting. MSCs were separated from human adipose tissue and cultured with HepG2 cell-derived Exosome(100 ng/nl)to initiate differentiation. The expressions of mesenchymal markers and several interleukins were also detected by Western blotting. HepG2 cells were co-cultured with the conditioned media(CM),in which HepG2 Exosome induced the differentiation of MSC into CAF. The expressions of epithelial and mesenchymal markers were detected by real-time polymerase chain reaction(PCR)and Western blotting. Cell proliferation was assessed using MTS assay. Transwell chambers were used in the in vitro migration and invasion assay. Results HepG2 cell-derived particles expressed CD63,70 kilodalton heat shock proteins,and 90 kilodalton heat shock proteins. With the treatment of HepG2 cell-derived Exosome,the expressions of mesenchymal marker α-smooth muscle actin,fibroblast activation protein α,interleukin(IL)-6,IL-8,and IL-1β were up-regulated,while vascular endothelial growth factor had no significant change. The conditioned media which HepG2 Exosome induced MSC differentiation CAF(CAF-CM)could significantly promote HepG2 cells proliferation(1.075±0.104),compared to BSA control(0.874±0.066,P=0.023)and MSC-CM(0.649±0.034,P=0.0005). CAF-CM could significantly enhance cell migration [(42.5±9.1) cells vs.(18.5±3.1) cells,P=0.001] and invasion [(29.0±3.5) cells vs.(13.1±3.7) cells,P=0.009] compared to its control group. Moreover the conditioned medium which HepG2 Exosome induced MSC to differentiate into CAF could also promote the expressions of mesenchyme-related genes Smad interacting protein 1(P=0.040),β-catenin(P=0.038),fibronectin(P=0.029),and Vimentin(P=0.013)and inhibit the expression of epithelial related genes zonula ocdudens-1(P=0.010).Conclusions Exosome extracted from HepG2 cells can induce human adipose-derived MSC to differentiate into cancer-associated myofibroblasts. CAF-like cells can promote the migration of the liver cancer cell line HepG2. |
|
来源
|
中国医学科学院学报
,2017,39(3):312-317 【核心库】
|
|
DOI
|
10.3881/j.issn.1000-503x.2017.03.003
|
|
关键词
|
Exosome
;
间充质干细胞
;
肝癌
;
肿瘤相关成纤维细胞
|
|
地址
|
1.
中国医学科学院北京协和医学院北京协和医院肿瘤内科, 北京, 100052
2.
中国医学科学院北京协和医学院基础医学研究所组织工程中心, 北京, 100005
|
|
语种
|
中文 |
|
文献类型
|
研究性论文 |
|
ISSN
|
1000-503X |
|
学科
|
肿瘤学 |
|
基金
|
国家自然科学基金
|
|
文献收藏号
|
CSCD:6021052
|
参考文献 共
15
共1页
|
|
1.
D'souza N. MSC and tumors:homing,differentiation,and secretion influence therapeutic potential.
Adv Biochem Eng Biotechnol,2013,130:209-266
|
CSCD被引
2
次
|
|
|
|
|
2.
Polanska U M. Carcinoma-associated fibroblasts:non-neoplastic tumour-promoting mesenchymal cells.
J Cell Physiol,2013,228(8):1651-1657
|
CSCD被引
10
次
|
|
|
|
|
3.
Paunescu V. Tumour-associated fibroblasts and mesenchymal stem cells:more similarities than differences.
J Cell Mol Med,2011,15(3):635-646
|
CSCD被引
8
次
|
|
|
|
|
4.
熊志勇. 肝癌细胞促进骨髓间充质干细胞向肿瘤相关成纤维细胞转化.
中华实验外科杂志,2015,32(8):1884-1887
|
CSCD被引
1
次
|
|
|
|
|
5.
Lee H K. Mesenchymal stem cells deliver exogenous miRNAs to neural cells and induce their differentiation and glutamate transporter expression.
Stem Cells Dev,2014,23(23):2851-2861
|
CSCD被引
11
次
|
|
|
|
|
6.
Chen T S. Measurement of precursor miRNA in Exosome from human ESC-derived mesenchymal stem cells.
Methods Mol Biol,2013,1024:69-86
|
CSCD被引
2
次
|
|
|
|
|
7.
Busch S. TGF-beta receptor type-2 expression in cancer-associated fibroblasts regulates breast cancer cell growth and survival and is a prognostic marker in pre-menopausal breast cancer.
Oncogene,2015,34(1):27-38
|
CSCD被引
5
次
|
|
|
|
|
8.
Haga H. Tumour cell-derived extracellular vesicles interact with mesenchymal stem cells to modulate the microenvironment and enhance cholangiocarcinoma growth.
J Extracell Vesicles,2015,4:24900
|
CSCD被引
9
次
|
|
|
|
|
9.
Chowdhury R. Cancer Exosome trigger mesenchymal stem cell differentiation into pro-angiogenic and pro-invasive myofibroblasts.
Oncotarget,2015,6(2):715-731
|
CSCD被引
10
次
|
|
|
|
|
10.
Ono M. Exosome from bone marrow mesenchymal stem cells contain a microRNA that promotes dormancy in metastatic breast cancer cells.
Sci Signal,2014,7(332):ra63
|
CSCD被引
25
次
|
|
|
|
|
11.
Gu J. Gastric cancer Exosome trigger differentiation of umbilical cord derived mesenchymal stem cells to carcinoma-associated fibroblasts through TGF-β/Smad pathway.
PLoS One,2012,7(12):e52465
|
CSCD被引
16
次
|
|
|
|
|
12.
Cho J A. Exosome from ovarian cancer cells induce adipose tissue-derived mesenchymal stem cells to acquire the physical and functional characteristics of tumor-supporting myofibroblasts.
Gynecol Oncol,2011,123(2):379-386
|
CSCD被引
15
次
|
|
|
|
|
13.
Kubo N. Cancer-associated fibroblasts in hepatocellular carcinoma.
World J Gastroenterol,2016,22(30):6841-6850
|
CSCD被引
10
次
|
|
|
|
|
14.
Ostman A. Cancer-associated fibroblasts and tumor growth-bystanders turning into key players.
Curr Opin Genet Dev,2009,19(1):67-73
|
CSCD被引
19
次
|
|
|
|
|
15.
Cirri P. Cancer-associated-fibroblasts and tumour cells:a diabolic liaison driving cancer progression.
Cancer Metastasis Rev,2012,31(1/2):195-208
|
CSCD被引
17
次
|
|
|
|
|
引证文献
1
篇
|
|
1
Tan Liping
Characteristics and regulation of mesenchymal stem cell plasticity by the microenvironment -specific factors involved in the regulation of MSC plasticity
Genes & Diseases,2022,9(2):296-309
CSCD被引
0
次
显示所有1篇文献
|
|
了解气象卫星更多信息，请访问