Role of apolipoprotein M in the proliferation,migration and invasion of non-small cell lung cancer A549 cells

doi:10.3969/j.issn.1673-8640.2020.09.019

Abstract

Abstract:

Objective To investigate the influence of apolipoprotein M（apo M） on the proliferation,migration and invasion of non-small cell lung cancer（NSCLC） A549 cell line and its relationship with matrix metalloproteinase（MMP）-10. Methods A549 cells being transfected with apo M lentivirus were used as apo M-OE group,and those being transfected with A549 empty lentivirus were used as negative control group. CCK-8 method,scratch test and Transwell invasion test were used to determine the proliferation,migration and invasion abilities. The expression levels of apo M and MMP-10 were determined by real-time fluorescence quantitative polymerase chain reaction（qRT-PCR） and immunoblotting. Results Compared with negative control group,the expression level of apo M in apo M-OE group was up-regulated（P<0.01）,and the expression level was about 28 times higher than that of negative control group. The proliferation and invasion rates of apo M-OE group were higher than those of negative control group（P<0.001）. Compared with negative control group,the blank field of vision in apo M-OE group was reduced,the closing rate was accelerated,and the migration rate was greater（P<0.001）. The expression levels of MMP-10 mRNA,apo M protein and MMP-10 protein in apo M-OE group were higher than those in negative control group（P<0.05）. Conclusions Over-expressing apo M can promote the proliferation,migration and invasion of A549 cells,and can make the cells highly expressing MMP-10.

Key words: Apolipoprotein M, Matrix metalloproteinase-10, Non-small cell lung cancer

CLC Number:

R446.1

WANG Zijin, ZHANG Xiaoying. Role of apolipoprotein M in the proliferation,migration and invasion of non-small cell lung cancer A549 cells[J]. Laboratory Medicine, 2020, 35(9): 937-942.

Figures/Tables 6

References 21

[1]	BRAY F,FERLAY J,SOERJOMATARAM I,et al.Global cancer statistics 2018:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin,2018,68(6):394-424.
[2]	FERLAY J,SOERJOMATARAM I,DIKSHIT R,et al.Cancer incidence and mortality worldwide:sources,methods and major patterns in GLOBOCAN 2012[J]. Int J Cancer,2015,136(5):E359-E386.
[3]	XU N,DAHLBÄCK B. A novel human apolipoprotein(apo M)[J]. J Biol Chem,1999,274(44):31286-31290.
[4]	TAKANO S,YOSHITOMI H,TOGAWA A,et al.Apolipoprotein C-1 maintains cell survival by preventing from apoptosis in pancreatic cancer cells[J]. Oncogene,2008,27(20):2810-2822.
[5]	SØILAND H,JANSSEN E A,KØRNER H,et al. Apolipoprotein D predicts adverse outcome in women >or=70 years with operable breast cancer[J]. Breast Cancer Res Treat,2009,113(3):519-528.
[6]	DIEPLINGER H,ANKERST D P,BURGES A,et al.Afamin and apolipoprotein A-IV:novel protein markers for ovarian cancer[J]. Cancer Epidemiol Biomarkers Prev,2009,18(4):1127-1133.
[7]	EGEBLAD M,WERB Z.New functions for the matrix metalloproteinases in cancer progression[J]. Nat Rev Cancer,2002,2(3):161-174.
[8]	BODEY B,BODEY B Jr,GRÖGER A M,et al. Invasion and metastasis:the expression and significance of matrix metalloproteinases in carcinomas of the lung[J]. In Vivo,2001,15(2):175-180.
[9]	JUSTILIEN V,REGALA R P,TSENG I C,et al.Matrix metalloproteinase-10 is required for lung cancer stem cell maintenance,tumor initiation and metastatic potential[J]. PLoS One,2012,7(4):e35040.
[10]	CHEN W,ZHENG R,BAADE P D,et al.Cancer statistics in China,2015[J]. CA Cancer J Clin,2016,66(2):115-132.
[11]	JIANG J,WU C,LUO G,et al.Expression of apolipoprotein M in human hepatocellular carcinoma tissues[J]. Acta Histochem,2011,113(1):53-57.
[12]	牟琴峰,罗光华,陈陆俊,等. 载脂蛋白M在结直肠癌组织中的表达及其临床病理相关性[J]. 中华胃肠外科杂志,2012,15(8):855-858.
[13]	REGALA R P,DAVIS R K,KUNZ A,et al.Atypical protein kinase C{iota} is required for bronchioalveolar stem cell expansion and lung tumorigenesis[J]. Cancer Res,2009,69(19):7603-7611.
[14]	ZHANG X,ZHU S,LUO G,et al.Expression of MMP-10 in lung cancer[J]. Anticancer Res,2007,27(4C):2791-2795.
[15]	ZHU Y,LUO G,JIANG B,et al.Apolipoprotein M promotes proliferation and invasion in non-small cell lung cancers via upregulating S1PR1 and activating the ERK1/2 and PI3K/AKT signaling pathways[J]. Biochem Biophys Res Commun,2018,501(2):520-526.
[16]	ZHOU Y,LI S,LI J,et al.Effect of microRNA-135a on cell proliferation,migration,invasion,apoptosis and tumor angiogenesis through the IGF-1/PI3K/Akt signaling pathway in non-small cell lung cancer[J]. Cell Physiol Biochem,2017,42(4):1431-1446.
[17]	ZHANG Y,HUANG L Z,YANG Q L,et al.Correlation analysis between ApoM gene-promoter polymorphisms and coronary heart disease[J]. Cardiovasc J Afr,2016,27(4):228-237.
[18]	MONTERO I,ORBE J,VARO N,et al.C-reactive protein induces matrix metalloproteinase-1 and -10 in human endothelial cells:implications for clinical and subclinical atherosclerosis[J]. J Am Coll Cardiol,2006,47(7):1369-1378.
[19]	ZHANG X,YIN P,DI D,et al.IL-6 regulates MMP-10 expression via JAK2/STAT3 signaling pathway in a human lung adenocarcinoma cell line[J]. Anticancer Res,2009,29(11):4497-4501.
[20]	黄波,罗光华,张晓膺,等. 载脂蛋白M对小鼠心肌炎症相关因子表达的影响[J]. 中华实验外科杂志,2017,34(4):641.
[21]	SRAMKOVA V,BEREND S,SIKLOVA M,et al.Apolipoprotein M:a novel adipokine decreasing with obesity and upregulated by calorie restriction[J]. Am J Clin Nutr,2019,109(6):1499-1510.

名称	序列（5´~3´）	长度/bp
apo M正义引物	TGCCCCGGAAATGGATCTA	60
apo M反义引物	CAGGGCGGCCTICAGTT	54
apo M探针	FAM-CACCTGACTGAAGGGAGCACAGATCTCA-TAMRA	87
MMP-10正义引物	TCGCCCAGTTCCGCCTT	54
MMP-10反义引物	GCACCAGGGGTTCCTCAGTAG	66
MMP-10探针	TGGCATTCAGTCTCTCTACGGACCTC	81
GAPDH正义引物	GGAAGGTGAAGGTCGGAGTC	63
GAPDH反义引物	CGTTCTCAGCCTTGACGGT	60
GAPDH探针	FAM-TTTGGTCGTATTGGGCGCCTG-TAMRA	66

名称	序列（5´~3´）	长度/bp
apo M正义引物	TGCCCCGGAAATGGATCTA	60
apo M反义引物	CAGGGCGGCCTICAGTT	54
apo M探针	FAM-CACCTGACTGAAGGGAGCACAGATCTCA-TAMRA	87
MMP-10正义引物	TCGCCCAGTTCCGCCTT	54
MMP-10反义引物	GCACCAGGGGTTCCTCAGTAG	66
MMP-10探针	TGGCATTCAGTCTCTCTACGGACCTC	81
GAPDH正义引物	GGAAGGTGAAGGTCGGAGTC	63
GAPDH反义引物	CGTTCTCAGCCTTGACGGT	60
GAPDH探针	FAM-TTTGGTCGTATTGGGCGCCTG-TAMRA	66