Role of long noncoding RNA-MALAT1 in human peritoneal mesothelial cells fibrosis induced with high glucose

doi:10.3877/cma.j.issn.2095-3216.2016.03.008

Abstract

Abstract:

Objective

To study the role of long noncoding RNA-MALAT1 in human peritoneal mesothelial cells (HPMCs) fibrosis induced with high glucose (HG).

Methods

The expression of MALAT1, E-cadherin, alpha-smooth muscle actin (ɑ-SMA), collagenⅠ, collagen Ⅲ, fibronectin, and connective tissue growth factor (CTGF) were tested by real-time PCR and Western blot in HPMCs incubated with HG medium (50 mmol/L glucose) or in control medium for different time (for 0, 1, 3, 5, and 7 days). The expression of MALAT1, E-cadherin, ɑ-SMA, collagenⅠ, collagenⅢ, fibronectin, and CTGF were also assayed by real-time PCR and Western blot in HPMCs after transfection with MALAT1 lentivirus, or control lentivirus. MALAT1 small interfering RNA (siRNA), or control siRNA was transfected into HPMCs that had been cultured with HG medium for 72 hours.

Results

The expression of MALAT1, ɑ-SMA, collagenⅠ, collagenⅢ, fibronectin, and CTGF in HPMCs cultured with HG medium for 72 hours were significantly increased (P<0.05), but E-cadherin decreased with the stimulation time of HG (P<0.05). Compared with control lentivirus, the expression of MALAT1, ɑ-SMA, collagenⅠ, collagenⅢ, fibronectin, and CTGF were increased, but E-cadherin was reduced after transfection with the MALAT1 lentivirus (P<0.01), suggesting that overexpression of MALAT1 might lead to an increase in epithelial-mesenchymal transition (EMT) and fibrosis of HPMCs. The expression of MALAT1, ɑ-SMA, collagen, collagenⅢ, fibronectin, and CTGF decreased (P<0.05), but E-cadherin increased (P<0.05) after silencing the expression of MALAT1 by siRNA compared with the control cells, demonstrating that down-regulation of MALAT1 could inhibit EMT and fibrosis in HG-induced HPMCs.

Conclusion

MALAT1 might participate in and promote the HG-induced fibrosis and damage of HPMCs.

Key words: Peritoneal fibrosis, Epithelial-mesenchymal transition, LncRNA, MALAT1

Tiantian Shi, Lijie He, Shiren Sun, Hanmin Wang. Role of long noncoding RNA-MALAT1 in human peritoneal mesothelial cells fibrosis induced with high glucose[J]. Chinese Journal of Kidney Disease Investigation(Electronic Edition), 2016, 05(03): 128-134.

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Figures/Tables 4

References 31

[1]	Garcia-Lopez E,Lindholm B,Davies S. An update on peritoneal dialysis solutions[J]. Nat Rev Nephrol, 2012, 8(4): 224-233.
[2]	Liem YS,Wong JB,Hunink MG, et al. Comparison of hemodialysis and peritoneal dialysis survival in The Netherlands[J]. Kidney Int, 2007, 71(2): 153-158.
[3]	Higuchi C,Nishimura H,Sanaka T. Biocompatibility of peritoneal dialysis fluid and influence of compositions on peritoneal fibrosis[J]. Ther Apher Dial, 2006, 10(4): 372-379.
[4]	Williams JD,Craig KJ,Topley N, et al. Morphologic changes in the peritoneal membrane of patients with renal disease[J]. J Am Soc Nephrol, 2002, 13(2): 470-479.
[5]	Witowski J,Wisniewska J,Korybalska K, et al. Prolonged exposure to glucose degradation products impairs viability and function of human peritoneal mesothelial cells[J]. J Am Soc Nephrol, 2001, 12(11): 2434-2441.
[6]	Morgan LW,Wieslander A,Davies M, et al. Glucose degradation products (GDP) retard remesothelialization independently of D-glucose concentration[J]. Kidney Int, 2003, 64(5): 1854-1866.
[7]	Mittelmaier S,Niwa T,Pischetsrieder M. Chemical and physiological relevance of glucose degradation products in peritoneal dialysis[J]. J Ren Nutr, 2012, 22(1): 181-185.
[8]	Yanez-Mo M,Lara-Pezzi E,Selgas R, et al. Peritoneal dialysis and epithelial-to-mesenchymal transition of mesothelial cells[J]. N Engl J Med, 2003, 348(5): 403-413.
[9]	Aroeira LS,Aguilera A,Sanchez-Tomero JA, et al. Epithelial to mesenchymal transition and peritoneal membrane failure in peritoneal dialysis patients: pathologic significance and potential therapeutic interventions[J]. J Am Soc Nephrol, 2007, 18(7): 2004-2013.
[10]	Krediet RT,Struijk DG. Peritoneal changes in patients on long-term peritoneal dialysis[J]. Nat Rev Nephrol, 2013, 9(7): 419-429.
[11]	Veneziano D,Nigita G,Ferro A. Computational approaches for the analysis of ncRNA through deep sequencing techniques[J]. Front Bioeng Biotechnol, 2015, 3: 77.
[12]	Mercer TR,Dinger ME,Mattick JS. Long non-coding RNAs: insights into functions[J]. Nat Rev Genet, 2009, 10(3): 155-159.
[13]	Batista PJ,Chang HY. Long noncoding RNAs: cellular address codes in development and disease[J]. Cell, 2013, 152(6): 1298-1307.
[14]	Yang L,Froberg JE,Lee JT. Long noncoding RNAs: fresh perspectives into the RNA world[J]. Trends Biochem Sci, 2014, 39(1): 35-43.
[15]	Hu L,Wu Y,Tan D, et al. Up-regulation of long noncoding RNA MALAT1 contributes to proliferation and metastasis in esophageal squamous cell carcinoma[J]. J Exp Clin Cancer Res, 2015, 34: 7.
[16]	Zhou S,Wang J,Zhang Z. An emerging understanding of long noncoding RNAs in kidney cancer[J]. J Cancer Res Clin Oncol, 2014, 140(12): 1989-1995.
[17]	Bernard D,Prasanth KV,Tripathi V, et al. A long nuclear-retained non-coding RNA regulates synaptogenesis by modulating gene expression[J]. EMBO J, 2010, 29(18): 3082-3093.
[18]	Ji P,Diederichs S,Wang W, et al. MALAT-1, a novel noncoding RNA, and thymosin beta4 predict metastasis and survival in early-stage non-small cell lung cancer[J]. Oncogene, 2003, 22(39): 8031-8041.
[19]	Hutchinson JN,Ensminger AW,Clemson CM, et al. A screen for nuclear transcripts identifies two linked noncoding RNAs associated with SC35 splicing domains[J]. BMC Genomics, 2007, 8: 39.
[20]	Lin R,Maeda S,Liu C, et al. A large noncoding RNA is a marker for murine hepatocellular carcinomas and a spectrum of human carcinomas[J]. Oncogene, 2007, 26(6): 851-858.
[21]	Sun R,Qin C,Jiang B, et al. Down-regulation of MALAT1 inhibits cervical cancer cell invasion and metastasis by inhibition of epithelial-mesenchymal transition[J]. Mol Biosyst, 2016, 12(3): 952-962.
[22]	Grassmann A,Gioberge S,Moeller S, et al. ESRD patients in 2004: global overview of patient numbers, treatment modalities and associated trends[J]. Nephrol Dial Transplant, 2005, 20(12): 2587-2593.
[23]	Moresco RN,Speeckaert MM,Delanghe JR. Diagnosis and monitoring of IgA nephropathy: the role of biomarkers as an alternative to renal biopsy[J]. Autoimmun Rev, 2015, 14(10): 847-853.
[24]	Fang XY,Pan HF,Leng RX, et al. Long noncoding RNAs: novel insights into gastric cancer[J]. Cancer Lett, 2015, 356(2 Pt B): 357-366.
[25]	Nakagawa S,Ip JY,Shioi G, et al. Malat1 is not an essential component of nuclear speckles in mice[J]. RNA, 2012, 18(8): 1487-1499.
[26]	Michalik KM,You X,Manavski Y, et al. Long noncoding RNA MALAT1 regulates endothelial cell function and vessel growth[J]. Circ Res, 2014, 114(9): 1389-1397.
[27]	Bamodu OA,Huang WC,Lee WH, et al. Aberrant KDM5B expression promotes aggressive breast cancer through MALAT1 overexpression and downregulation of hsa-miR-448[J]. BMC Cancer, 2016, 16: 160.
[28]	Xiao H,Tang K,Liu P, et al. LncRNA MALAT1 functions as a competing endogenous RNA to regulate ZEB2 expression by sponging miR-200s in clear cell kidney carcinoma[J]. Oncotarget, 2015, 6(35): 38005-38015.
[29]	Feng T,Shao F,Wu Q, et al. miR-124 downregulation leads to breast cancer progression via LncRNA-MALAT1 regulation and CDK4/E2F1 signal activation[J]. Oncotarget, 2016, 7(13): 16205-16216.
[30]	Ma J,Wang P,Yao Y, et al. Knockdown of long non-coding RNA MALAT1 increases the blood-tumor barrier permeability by up-regulating miR-140[J]. Biochim Biophys Acta, 2016, 1859(2): 324-338.
[31]	Salmena L,Poliseno L,Tay Y, et al. A ceRNA hypothesis: the Rosetta Stone of a hidden RNA language?[J]. Cell, 2011, 146(3): 353-358.

基因	引物序列
β-actin	正义引物：5′-CTCCATCCTGGCCTCGCTGT-3′
β-actin	反义引物：5′-GCTGTCACCTTCACCGTTCC-3′
α-SMA	正义引物：5′-TCCGGAGCGCAAATACTCTG-3′
α-SMA	反义引物：5′-CCGGCTTCATCGTATTC-3′
CTGF	正义引物：5′-CTCTGCTCTCGCAGCTTACC-3′
CTGF	反义引物：5′-CATCCCACAGGTCTTGGAAC-3′
ColⅠ	正义引物：5′-TGACCTCAAGATGTGCCACT-3′
ColⅠ	反义引物：5′-CCTCTTGTCCTTGGGGTTCT-3′
ColⅢ	正义引物：5′-CGAAACACACTGGGGAATGG-3′
ColⅢ	反义引物：5′-TTTGTCGGTCACTTGCACTG-3′
Fn	正义引物：5′-GGACATGCATTGCCTACTCG-3′
Fn	反义引物：5′-GAATCCTGGCATTGGTCGAC-3′
E-cad	正义引物：5′-CGCATTGCCACATACACTCT-3′
E-cad	反义引物：5′-TTGGCTGAGGATGGTGTAAG-3′
MALAT1	正义引物：5′- CTAGCTAGCGCTTACAATCTTAGAGTGGTAGGCA -3′
MALAT1	反义引物：5′- CCGCTCGAGTCTAACATAGTTCAACCCACCAAAG-3′

基因	引物序列
β-actin	正义引物：5′-CTCCATCCTGGCCTCGCTGT-3′
β-actin	反义引物：5′-GCTGTCACCTTCACCGTTCC-3′
α-SMA	正义引物：5′-TCCGGAGCGCAAATACTCTG-3′
α-SMA	反义引物：5′-CCGGCTTCATCGTATTC-3′
CTGF	正义引物：5′-CTCTGCTCTCGCAGCTTACC-3′
CTGF	反义引物：5′-CATCCCACAGGTCTTGGAAC-3′
ColⅠ	正义引物：5′-TGACCTCAAGATGTGCCACT-3′
ColⅠ	反义引物：5′-CCTCTTGTCCTTGGGGTTCT-3′
ColⅢ	正义引物：5′-CGAAACACACTGGGGAATGG-3′
ColⅢ	反义引物：5′-TTTGTCGGTCACTTGCACTG-3′
Fn	正义引物：5′-GGACATGCATTGCCTACTCG-3′
Fn	反义引物：5′-GAATCCTGGCATTGGTCGAC-3′
E-cad	正义引物：5′-CGCATTGCCACATACACTCT-3′
E-cad	反义引物：5′-TTGGCTGAGGATGGTGTAAG-3′
MALAT1	正义引物：5′- CTAGCTAGCGCTTACAATCTTAGAGTGGTAGGCA -3′
MALAT1	反义引物：5′- CCGCTCGAGTCTAACATAGTTCAACCCACCAAAG-3′

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