High SIPA-1 expression in proximal tubules of human kidneys under pathological conditions

Abstract

Systemic lupus erythematosus (SLE) and clear cell renal cell carcinoma (CC-RCC) are serious disorders and usually fatal, and always accompanied with pathological changes in the kidney. Signal-induced proliferation-associated protein 1 (SIPA-1) is a Rap1GTPase activating protein (Rap1GAP) expressed in the normal distal and collecting tubules of the murine kidney. Lupus-like autoimmune disease and leukemia have been observed in SIPA-1 deficient mice, suggesting a pathological relevance of SIPA-1 to SLE and carcinoma in human being. The expression pattern of SIPA-1 is as yet undefined and the pathogenesis of these diseases in humans remains elusive. In this study, we used both immunohistochemistry and quantum dot (QD)-based immunofluorescence staining to investigate the expression of SIPA-1 in renal specimens from SLE and CC-RCC patients. MTT assay and Western blotting were employed to evaluate the effects of SIPA-1 overexpression on the proliferation and apoptosis of renal cell lines. Semi-quantitative reverse transcriptase-PCR (RT-PCR) was applied to examine the changes of hypoxia-inducible factor-1α (HIF-1α) mRNA level. Results showed that SIPA-1 was highly expressed in the proximal and collecting tubules of nephrons in SLE patients compared to normal ones, and similar results were obtained in the specimens of CC-RCC patients. Although SIPA-1 overexpression did not affect cellular proliferation and apoptosis of both human 786-O renal cell carcinoma cells and rat NRK-52E renal epithelial cell lines, RT-PCR results showed that HIF-1α mRNA level was down-regulated by SIPA-1 overexpression in 786-O cells. These findings suggest that SIPA-1 may play critical roles in the pathological changes in kidney, and might provide a new biomarker to aid in the diagnosis of SLE and CC-RCC.

Keywords

immunohistochemistry / signal-induced proliferation-associated protein 1 / hypoxia-inducible factor-1α / systemic lupus erythematosus / clear cell renal cell carcinoma

References

[1] CrispinJC, LiossisSN, Kis-TothK, et al. . Pathogenesis of human systemic lupus erythematosus: recent advances. Trends Mol Med, 2010, 16(2): 47-57 PMCID: 2823952 PMID: 20138006

[2] ManoharanA, MadaioMP. Biomarkers in lupus nephritis. Rheum Dis Clin North Am, 2010, 36(1): 131-143 PMID: 20202595

[3] SuhCH, KimHA. Cytokines and their receptors as biomarkers of systemic lupus erythematosus. Expert Rev Mol Diagn, 2008, 8(2): 189-198 PMID: 18366305

[4] BadotV, LuijtenRK, van RoonJA, et al. . Serum soluble interleukin 7 receptor is strongly associated with lupus nephritis in patients with systemic lupus erythematosus. Ann Rheum Dis, 2013, 72(3): 453-456 PMID: 23264357

[5] PradhanV, MahantG, RajadhyakshaA, et al. . A study on anti-mannose binding lectin (anti-MBL) antibodies and serum MBL levels in Indian systemic lupus erythematosus patients. Rheumatol Int, 2013, 33(6): 1533-1539 PMID: 23255139

[6] ChamCM, KoK, NiewoldTB. Interferon regulatory factor 5 in the pathogenesis of systemic lupus erythematosus. Clin Dev Immunol, 2012, 2012: 780436 PMCID: 3509422 PMID: 23251221

[7] NodaY, HorikawaS, FurukawaT, et al. . Aquaporin-2 trafficking is regulated by PDZ-domain containing protein SPA-1. FEBS Lett, 2004, 568(1–3): 139-145 PMID: 15196935

[8] IshidaD, SuL, TamuraA, et al. . Rap1 signal controls B cell receptor repertoire and generation of self-reactive B1a cells. Immunity, 2006, 24(4): 417-427 PMID: 16618600

[9] IshidaD, KometaniK, YangH, et al. . Myeloproliferative stem cell disorders by deregulated Rap1 activation in SPA-1-deficient mice. Cancer Cell, 2003, 4(1): 55-65 PMID: 12892713

[10] ParkYG, ZhaoX, LesueurF, et al. . Sipa1 is a candidate for underlying the metastasis efficiency modifier locus Mtes1. Nat Genet, 2005, 37(10): 1055-1062 PMCID: 2140048 PMID: 16142231

[11] CrawfordNP, ZiogasA, PeelDJ, et al. . Germline polymorphisms in SIPA1 are associated with metastasis and other indicators of poor prognosis in breast cancer. Breast Cancer Res, 2006, 8(2): R16 PMCID: 1483843 PMID: 16563182

[12] ShimizuY, HamazakiY, HattoriM, et al. . SPA-1 controls the invasion and metastasis of human prostate cancer. Cancer Sci, 2011, 102(4): 828-836 PMID: 21251160

[13] VirtanenI, LehtoVP. Progression of malignancy in clear cell renal cell carcinomas. Scand J Surg, 2004, 93(2): 112-117 PMID: 15285562

[14] YeungRS, XiaoGH, JinF, et al. . Predisposition to renal carcinoma in the Eker rat is determined by germ-line mutation of the tuberous sclerosis 2 (TSC2) gene. Proc Natl Acad Sci USA, 1994, 91(24): 11 413-11 416

[15] KimWJ, GerseyZ, DaakaY. Rap1GAP regulates renal cell carcinoma invasion. Cancer Lett, 2012, 320(1): 65-71 PMCID: 3319804 PMID: 22266190

[16] WuJ, ZhangY, FrilotN, et al. . Prostaglandin E2 regulates renal cell carcinoma invasion through the EP4 receptor-Rap GTPase signal transduction pathway. J Biol Chem, 2011, 286(39): 33 954-33 962

[17] HaaseVH. The VHL/HIF oxygen-sensing pathway and its relevance to kidney disease. Kidney Int, 2006, 69(8): 1302-1307 PMID: 16531988

[18] ZhouL, YangH. The von Hippel-Lindau tumor suppressor protein promotes c-Cbl-independent poly-ubiquitylation and degradation of the activated EGFR. PLoS One, 2011, 6(9): e23 936

[19] LinehanWM, BratslavskyG, PintoPA, et al. . Molecular diagnosis and therapy of kidney cancer. Annu Rev Med, 2010, 61: 329-343 PMCID: 2921612 PMID: 20059341

[20] TanEM, CohenAS, FriesJF, et al. . The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum, 1982, 25(11): 1271-1277 PMID: 7138600

[21] ChenC, PengJ, XiaHS, et al. . Quantum dots-based immunofluorescence technology for the quantitative determination of HER2 expression in breast cancer. Biomaterials, 2009, 30(15): 2912-2918 PMID: 19251316

[22] TholouliE, SweeneyE, BarrowE, et al. . Quantum dots light up pathology. J Pathol, 2008, 216(3): 275-285 PMID: 18814189

[23] ChetramMA, HintonCV. PTEN regulation of ERK1/2 signaling in cancer. J Recept Signal Transduct Res, 2012, 32(4): 190-195 PMCID: 3503241 PMID: 22737980

[24] KaelinWJ. The von Hippel-Lindau tumour suppressor protein: O2 sensing and cancer. Nat Rev Cancer, 2008, 8(11): 865-873 PMID: 18923434

[25] MathieuV, PirkerC, SchmidtWM, et al. . Aggressiveness of human melanoma xenograft models is promoted by aneuploidy-driven gene expression deregulation. Oncotarget, 2012, 3(4): 399-413 PMCID: 3380575 PMID: 22535842

[26] CarmelietP, JainRK. Angiogenesis in cancer and other diseases. Nature, 2000, 407(6801): 249-257 PMID: 11001068

[27] BertoutJA, PatelSA, SimonMC. The impact of O2 availability on human cancer. Nat Rev Cancer, 2008, 8(12): 967-975 PMCID: 3140692 PMID: 18987634

[28] CockmanME, MassonN, MoleDR, et al. . Hypoxia inducible factor-alpha binding and ubiquitylation by the von Hippel-Lindau tumor suppressor protein. J Biol Chem, 2000, 275(33): 25 733-25 741

[29] TalksKL, TurleyH, GatterKC, et al. . The expression and distribution of the hypoxia-inducible factors HIF-1alpha and HIF-2alpha in normal human tissues, cancers, and tumor-associated macrophages. Am J Pathol, 2000, 157(2): 411-421 PMCID: 1850121 PMID: 10934146