孤儿核受体SHP对人骨肉瘤细胞株143B的影响及机制
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摘要
目的分析孤儿核受体小异二聚体伴侣(small heterodimer partner,SHP)对人骨肉瘤细胞株143B增殖、凋亡、迁移和侵袭的影响,并初步探讨其可能机制。方法分别构建过表达SHP的腺病毒、以及SHP的RNA干扰(RNA interference,RNAi)腺病毒;结晶紫染色和克隆形成实验检测143B细胞增殖情况;Horchest33258染色检测143B细胞凋亡情况;Transwell实验检测143B细胞迁移和细胞侵袭情况;Western Blot检测143B细胞Wnt/β-catenin信号的活化情况。结果过表达SHP可抑制143B细胞增殖、迁移和侵袭(P<0.01),但促进细胞凋亡(P<0.01);反之,抑制SHP表达,可促进143B细胞增殖、迁移和侵袭(P<0.01),但对细胞凋亡无明显影响。过表达SHP可降低β-catenin及其下游靶分子Cyclin D1和c-Myc的蛋白水平(P<0.01);抑制SHP表达可增加β-catenin、Cyclin D1和c-Myc的蛋白水平(P<0.01)。结论 SHP可抑制143B细胞增殖、迁移和侵袭,并促进细胞凋亡,这可能与SHP抑制Wnt/β-catenin信号活性有关。
Objective To analyze the effect of orphan nuclear receptor small heterodimer partner(SHP) on the proliferation,apoptosis,migration and invasion of 143 B human osteosarcoma cell line,and to explore its possible mechanism of action.Methods The SHP-overexpressing adenovirus vectors and RNA interference-carrying adenovirus vectors were constructed in this study.Cell proliferation was determined by crystal violet staining and colony formation assay.Cell apoptosis was detected by Horchest 33258 staining.Cell migration and invasion were analyzed by transwell assay.The activation of Wnt/β-catenin signal was determined by Western blot.Results The overexpression of SHP inhibited the proliferation,migration and invasion but promoted the apoptosis in 143 B cells(P<0.01).On the contrary,the inhibition of SHP expression promoted the proliferation,migration and invasion,but had no obvious effect on the apoptosis in 143 B cells.Furthermore,the overexpression of SHP reduced the protein expression of β-catenin and its downstream target molecules cyclin D1 and c-Myc(P<0.01).The inhibition of SHP expression increased the protein levels of β-catenin,cyclin D1 and c-Myc(P<0.01).Conclusion SHP may suppress the proliferation,apoptosis and invasion and promote the apoptosis in 143 B cells partially through the inhibition of Wnt/β-catenin signal.
Objective To analyze the effect of orphan nuclear receptor small heterodimer partner(SHP) on the proliferation,apoptosis,migration and invasion of 143 B human osteosarcoma cell line,and to explore its possible mechanism of action.Methods The SHP-overexpressing adenovirus vectors and RNA interference-carrying adenovirus vectors were constructed in this study.Cell proliferation was determined by crystal violet staining and colony formation assay.Cell apoptosis was detected by Horchest 33258 staining.Cell migration and invasion were analyzed by transwell assay.The activation of Wnt/β-catenin signal was determined by Western blot.Results The overexpression of SHP inhibited the proliferation,migration and invasion but promoted the apoptosis in 143 B cells(P<0.01).On the contrary,the inhibition of SHP expression promoted the proliferation,migration and invasion,but had no obvious effect on the apoptosis in 143 B cells.Furthermore,the overexpression of SHP reduced the protein expression of β-catenin and its downstream target molecules cyclin D1 and c-Myc(P<0.01).The inhibition of SHP expression increased the protein levels of β-catenin,cyclin D1 and c-Myc(P<0.01).Conclusion SHP may suppress the proliferation,apoptosis and invasion and promote the apoptosis in 143 B cells partially through the inhibition of Wnt/β-catenin signal.
引文
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[3] HELMAN L J,MELTZER P.Mechanisms of sarcoma development[J].Nat Rev Cancer,2003,3(9):685-694.
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[5] DENDULURI S K,IDOWU O,WANG Z,et al.Insulin-like growth factor(IGF)signaling in tumorigenesis and the development of cancer drug resistance[J].Genes Dis,2015,2(1):13-25.
[6] HAYDON R C,DEYRUP A,ISHIKAWA A,et al.Cytoplasmic and/or nuclear accumulation of the beta-catenin protein is a frequent event in human osteosarcoma[J].Int J Cancer,2002,102(4):338-342.
[7] WANG R N,GREEN J,WANG Z L,et al.Bone morphogenetic protein(BMP) signaling in development and human diseases[J].Genes Dis,2014,1(1):87-105.
[8] MCEWAN I J.The nuclear receptor superfamily at thirty[J].Methods Mol Biol,2016,1443:3-9.
[9] ZHANG Y X,HAGEDORN C H,WANG L.Role of nuclear receptor SHP in metabolism and cancer[J].Biochim Biophys Acta,2011,1812(8):893-908.
[10] ZOU A,LEHN S,MAGEE N,et al.New insights into orphan nuclear receptor SHP in liver cancer [J].Nucl Receptor Res,2015,2.pii:101162.
[11] ZHANG Y X,SOTO J,PARK K,et al.Nuclear receptor SHP,a death receptor that targets mitochondria,induces apoptosis and inhibits tumor growth[J].Mol Cell Biol,2010,30(6):1341-1356.
[12] HEDRICK E,LEE S O,SAFE S.The nuclear orphan receptor NR4A1 regulates beta 1-integrin expression in pancreatic and colon cancer cells and can be targeted by NR4A1 antagonis-ts[J].Mol Carcinog,2017,56(9):2066-2075.
[13] JIANG H L,XU D,YU H,et al.DAX-1 inhibits hepatocellular carcinoma proliferation by inhibiting β-catenin transcriptional activity[J].Cell Physiol Biochem,2014,34(3):734-742.
[14] YANG Z H,KOEHLER A N,WANG Li.A novel small molecule activator of nuclear receptor SHP inhibits HCC cell migration via suppressing Ccl2[J].Mol Cancer Ther,2016,15(10):2294-2301.
[15] KHORASANI M,TEIMOORI TOOLABI L,FARIVAR T N,et al.Aberrant expression of miR-141 and nuclear receptor small heterodimer partner in clinical samples of prostate cancer[J].Cancer Biomark,2018,22(1):19-28.
[16] WU D,CHEUNG A,WANG Y,et al.The emerging roles of orphan nuclear receptors in prostate cancer[J].Biochim Biophys Acta,2016,1866(1):23-36.
[17] LI G D,KONG B,ZHU Y,et al.Small heterodimer partner overexpression partially protects against liver tumor development in farnesoid X receptor knockout mice[J].Toxicol Appl Pharmacol,2013,272(2):299-305.
[18] HANAHAN D,WEINBERG R A.Hallmarks of cancer:the next Generation[J].Cell,2011,144(5):646-674.
[19] YI X J,ZHAO Y H,QIAO L X,et al.Aberrant Wnt/β-catenin signaling and elevated expression of stem cellproteins are associated with osteosarcoma side population cells of high tumorigenicity[J].Mol Med Rep,2015,12(4):5042-5048.