MFN1在肝癌转移中的调控作用研究
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摘要
目的:探讨线粒体融合蛋白MFN1(mito-fusion 1)在肝癌转移中的作用及其机制。方法:1).采用免疫组化实验检测15对肝癌转移灶组织与原发灶组织中MFN1的表达,以明确肝癌转移时是否伴有MFN1表达的改变。2).采用si RNA (small interference RNA)下调肝癌细胞中MFN1的表达后,提高Transwell迁移实验和Transwell侵袭实验分别检测其迁移和侵袭能力,通过实时荧光定量PCR (Quantitative Real-time PCR,qRT-PCR)和Western blot实验分别检测基质金属蛋白酶1 (matrix metalloproteinase 1,MMP1)、MMP2、MMP7及MMP9的m RNA和蛋白表达。结果:1)肝癌转移灶组织中MFN1表达显著低于原发灶组织(P<0.05)。2).下调MFN1表达后,肝癌细胞的迁移和侵袭能力显著升高,MMP7的表达显著增加,而MMP1、MMP2与MMP9的表达无明显变化。结论:线粒体融合蛋白MFN1在肝癌转移组织中表达显著降低,可能通过激活MMP7表达,促进肝癌细胞侵袭和转移。
Objective: To investigate the role of the mitochondrial fusion MFN1 in the metastasis of hepatocellular carcinoma(HCC) cells. Methods: 1. Immunohistochemistry assay was used for evaluating the expression of MFN1 in 15 paired metastatic and primary tissues of HCC to determine whether the expression of MFN1 was changed during metastasis. 2. After MFN1 was knocked-down in HCC cells by small interference RNA(si RNA), transwell migration and invasion assays were used to analyze the migration and invasion capabilities of HCC cells. qRT-PCR and western blot analysis were used for detection of expression levels of matrix metalloproteinase(MMPs) of MMP1, MMP2, MMP7 and MMP9. Results: The expression of MFN1 in metastatic tissues of HCC is significantly lower than their paired primary tissues. Knockdown of MFN1 upregulated the migration and invasion abilities, and the expression of MMP7 in HCC cells, while had no effect on the expressions of MMP1, MMP2 and MMP9. Conclusions: The expression of mitochondrial fission factor MFN1 is downregulated in metastatic tissues, which may promote the invasion and metastasis of HCC cells by upregulating of MMP7 expression.
Objective: To investigate the role of the mitochondrial fusion MFN1 in the metastasis of hepatocellular carcinoma(HCC) cells. Methods: 1. Immunohistochemistry assay was used for evaluating the expression of MFN1 in 15 paired metastatic and primary tissues of HCC to determine whether the expression of MFN1 was changed during metastasis. 2. After MFN1 was knocked-down in HCC cells by small interference RNA(si RNA), transwell migration and invasion assays were used to analyze the migration and invasion capabilities of HCC cells. qRT-PCR and western blot analysis were used for detection of expression levels of matrix metalloproteinase(MMPs) of MMP1, MMP2, MMP7 and MMP9. Results: The expression of MFN1 in metastatic tissues of HCC is significantly lower than their paired primary tissues. Knockdown of MFN1 upregulated the migration and invasion abilities, and the expression of MMP7 in HCC cells, while had no effect on the expressions of MMP1, MMP2 and MMP9. Conclusions: The expression of mitochondrial fission factor MFN1 is downregulated in metastatic tissues, which may promote the invasion and metastasis of HCC cells by upregulating of MMP7 expression.
引文
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[2]Lee MS.Role of mitochondrial function in cell death and body metabolism[J].Frontiers in bioscience,2016,21(1):1233-1244
[3]Picard M,Taivassalo T,Gouspillou G,et al.Mitochondria:isolation,structure and function[J].The Journal of physiology,2011,589(18):4413-4421
[4]Friedman JR,Nunnari J.Mitochondrial form and function[J].Nature,2014,505(7483):335-343
[5]Westermann B.Mitochondrial fusion and fission in cell life and death[J].Nature reviews Molecular cell biology,2010,11(12):872-884
[6]Devoto VMP,Falzone TL.Mitochondrial dynamics in Parkinson's disease:a role for alpha-synuclein[J].Dis Model Mech,2017,10(9):1075-1087
[7]Mishra P.Interfaces between mitochondrial dynamics and disease[J].Cell Calcium,2016,60(3):190-198
[8]Gao J,Wang L,Liu J,et al.Abnormalities of Mitochondrial Dynamics in Neurodegenerative Diseases[J].Antioxidants,2017,6(2):25-33
[9]Marin-Garcia J,Akhmedov AT.Mitochondrial dynamics and cell death in heart failure[J].Heart failure reviews,2016,21(2):123-136
[10]Mishra P.Interfaces between mitochondrial dynamics and disease[J].Cell Calcium,2016,60(3):190-198
[11]Rovira-Llopis S,Banuls C,Diaz-Morales N,et al.Mitochondrial dynamics in type 2 diabetes:Pathophysiological implications[J].Redox Biol,2017,11(1):637-645
[12]Srinivasan S,Guha M,Kashina A,et al.Mitochondrial dysfunction and mitochondrial dynamics-The cancer connection[J].Biochimica et biophysica acta,2017,1858(8):602-614
[13]Trotta AP,Chipuk JE.Mitochondrial dynamics as regulators of cancer biology[J].Cellular and molecular life sciences,2017,74(11):1999-2017
[14]Rehman J,Zhang HJ,Toth PT,et al.Inhibition of mitochondrial fission prevents cell cycle progression in lung cancer[J].FASEB journal,2012,26(5):2175-2186
[15]Kim YY,Yun SH,Yun J.Downregulation of Drp1,a fission regulator,is associated with human lung and colon cancers[J].Acta biochimica et biophysica Sinica,2018,50(2):209-215
[16]Jia Y,Zhou L,Tian C,et al.Dynamin-related protein 1 is involved in micheliolide-induced breast cancer cell death[J].Onco Targets Ther,2015,8(1):3371-3381
[17]Jablonska-Trypuc A,Matejczyk M,Rosochacki S.Matrix metalloproteinases(MMPs),the main extracellular matrix(ECM)enzymes in collagen degradation,as a target for anticancer drugs[J].Journal of enzyme inhibition and medicinal chemistry,2016,31(1):177-183
[18]Zhong Y,Lu YT,Sun Y,et al.Recent opportunities in matrix metalloproteinase inhibitor drug design for cancer[J].Expert Opin Drug Dis,2018,13(1):75-87
[19]Yadav L,Puri N,Rastogi V,et al.Matrix metalloproteinases and cancer-roles in threat and therapy[J].Asian Pacific journal of cancer prevention,2014,15(3):1085-1091
[20]Jafri MS,Kumar R.Modeling mitochondrial function and its role in disease[J].Progress in molecular biology and translational science,2014,123(1):103-125
[21]Craven L,Alston CL,Taylor RW,et al.Recent Advances in Mitochondrial Disease[J].Annual review of genomics and human genetics,2017,18(1):257-275
[22]Alston CL,Rocha MC,Lax NZ,et al.The genetics and pathology of mitochondrial disease[J].The Journal of pathology,2017,241(2):236-250
[23]Rolfo A,Pedra LL,Todros T,et al.Dynamic Role of Noxa in Regulating Mitochondrial Cell Death and Autophagy in the Human Placenta[J].Reprod Sci,2012,19(S3):85a-85a(下转第1612页)
[24]Haroon S,Vermulst M.Linking mitochondrial dynamics to mitochondrial protein quality control[J].Current opinion in genetics&development,2016,38(1):68-74
[25]Ni HM,Williams JA,Ding WX.Mitochondrial dynamics and mitochondrial quality control[J].Redox Biol,2015,4(1):6-13
[26]Trotta AP,Chipuk JE.Mitochondrial dynamics as regulators of cancer biology[J].Cellular and molecular life sciences,2017,74(11):1999-2017
[27]Inoue-Yamauchi A,Oda H.Depletion of mitochondrial fission factor DRP1 causes increased apoptosis in human colon cancer cells[J].Biochem Biophys Res Commun,2012,421(1):81-85
[28]Rehman J,Zhang HJ,Toth PT,et al.Inhibition of mitochondrial fission prevents cell cycle progression in lung cancer[J].Faseb Journal2012,26(5):2175-2186
[29]Kashatus JA,Nascimento A,Myers LJ,et al.Erk2 Phosphorylation of Drp1 Promotes Mitochondrial Fission and MAPK-Driven Tumor Growth[J].Mol Cell,2015,57(3):537-551
[30]Huang QC,Zhan L,Cao HY,et al.Increased mitochondrial fission promotes autophagy and hepatocellular carcinoma cell survival through the ROS-modulated coordinated regulation of the NFKB and TP53 pathways[J].Autophagy,2016,12(6):999-1014
[31]Zhan L,Cao HY,Wang G,et al.Drp1-mediated mitochondrial fission promotes cell proliferation through crosstalk of p53 and NF-kappa Bpathways in hepatocellular carcinoma[J].Oncotarget,2016,7(40):65001-65011
[32]Zhao J,Zhang J,Yu M,et al.Mitochondrial dynamics regulates migration and invasion of breast cancer cells[J].Oncogene,2013,32(40):4814-4824
[33]Che TF,Lin CW,Wu YY,et al.Mitochondrial translocation of EGFRregulates mitochondria dynamics and promotes metastasis in NSCLC[J].Oncotarget,2015,6(35):37349-37366
[34]Sun X,Cao H,Zhan L,et al.Mitochondrial fission promotes cell migration by Ca(2+)/Ca MKII/ERK/FAK pathway in hepatocellular carcinoma[J].Liver international,2018,38(7):1263-1272
[35]Kang KF,Zhang X,Chen XW.Expressions of MDM2 and MMP-7 in normal liver,cirrhosis,hepatocellular carcinoma tissues[J].Chinese journal of hepatology,2010,18(1):67-68
[36]Chen L,Li M,Li Q,et al.DKK1 promotes hepatocellular carcinoma cell migration and invasion through beta-catenin/MMP7 signaling pathway[J].Molecular cancer,2013,12:157
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