Elevated expression of SLC34A2 inhibits the viability and invasion of A549 cells

Yang,Weihan; Wang,Yu; Pu,Qiang; Ye,Sujuan; Ma,Qingping; Ren,Jiang; Zhong,Guoxing; Liu,Lunxu; Zhu,Wen

doi:10.3892/mmr.2014.2376

Elevated expression of SLC34A2 inhibits the viability and invasion of A549 cells

Authors:
- Weihan Yang
- Yu Wang
- Qiang Pu
- Sujuan Ye
- Qingping Ma
- Jiang Ren
- Guoxing Zhong
- Lunxu Liu
- Wen Zhu
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Affiliations: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China, Department of Thoracic Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China
Published online on: July 14, 2014 https://doi.org/10.3892/mmr.2014.2376
Pages: 1205-1214
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Abnormal expression of solute carrier family 34 (sodium phosphate), member 2 (SLC34A2) in the lung may induce abnormal alveolar type II (AT II) cells to transform into lung adenocarcinoma cells, and may also be important in biological process of lung adenocarcinoma. However, at present, the effects and molecular mechanisms of SLC34A2 in the initiation and progression of lung cancer remain to be elucidated. To the best of our knowledge, the present study revealed for the first time that the expression levels of SLC34A2 were downregulated in the A549 and H1299 lung adenocarcinoma cell lines. Further investigation demonstrated that the elevated expression of SLC34A2 in A549 cells was able to significantly inhibit cell viability and invasion in vitro. In addition, 10 upregulated genes between the A549‑P‑S cell line stably expressing SLC34A2 and the control cell line A549‑P were identified by microarray analysis and quantitative polymerase chain reaction, including seven tumor suppressor genes and three complement genes. Furthermore, the upregulation of complement gene C3 and complement 4B preproprotein (C4b) in A549‑P‑S cells was confirmed by ELISA analysis and was identified to be correlated with recovering Pi absorption in A549 cells by the phosphomolybdic acid method by enhancing the expression of SLC34A2. Therefore, it was hypothesized that the mechanisms underlying the effect of SLC34A2 on A549 cells might be associated with the activation of the complement alternative pathway (C3 and C4b) and upregulation of the expression of selenium binding protein 1, thioredoxin‑interacting protein, PDZK1‑interacting protein 1 and dual specificity protein phosphatase 6. Downregulation of SLC34A2 may primarily cause abnormal AT II cells to escape from complement‑associated immunosurveillance and abnormally express certain tumor‑suppressor genes inducing AT II cells to develop into lung adenocarcinoma. The present study further elucidated the effects and mechanisms of SLC34A2 in the generation and development of lung cancer.

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1	Jemal A, Siegel R, Ward E, Hao Y, Xu J and Thun MJ: Cancer statistics, 2009. CA Cancer J Clin. 59:225–249. 2009. View Article : Google Scholar
2	Murer H, Forster I and Biber J: The sodium phosphate cotransporter family SLC34. Pfugers Arch. 447:763–767. 2004. View Article : Google Scholar : PubMed/NCBI
3	Zheng X, Kammerer CM, Cox LA, Morrison A, Turner ST and Ferrell RE: Association of SLC34A2 variation and sodium-lithium countertransport activity in humans and baboons. Am J Hypertens. 22:288–293. 2009. View Article : Google Scholar : PubMed/NCBI
4	Lituiev DS and Kiyamova RG: Mutations in the gene of human type IIb sodium-phosphate cotransporter SLC34A2. Biopolym Cell. 26:13–22. 2010. View Article : Google Scholar
5	Tenenhouse HS: Phosphate transport: Molecular basis, regulation and pathophysiology. J Steroid Biochem Mol Biol. 103:572–577. 2007. View Article : Google Scholar : PubMed/NCBI
6	Xu H, Bai L, Collins JF and Ghishan FK: Molecular cloning, functional characterization, tissue distribution, and chromosomal localization of a human, small intestinal sodium-phosphate (Na+-Pi) transporter (SLC34A2). Genomics. 62:281–284. 1999. View Article : Google Scholar
7	Corut A, Senyigit A, Ugur S, Altin S, Ozcelik U, Calisir H, Yildirim Z, Gocmen A and Tolun A: Mutations in SLC34A2 cause pulmonary alveolar microlithiasis and are possibly associated with testicular microlithiasis. Am J Hum Genet. 79:650–656. 2006. View Article : Google Scholar
8	Vachon L, Fareau GE, Wilson MG and Chan LS: Testicular microlithiasis in patients with Down syndrome. J Pediatr. 149:233–236. 2006. View Article : Google Scholar : PubMed/NCBI
9	Hernando N, Sheikh S, Karim-Jimenez Z, Galliker H, Forgo J and Biber J: Asymmetrical targeting of type II Na-P(i) cotransporters in renal and intestinal epithelial cell lines. Am J Physiol Renal Physiol. 278:F361–F368. 2000.PubMed/NCBI
10	Cerri MF, Rezende LC, Paes FM, Silva IV and Rangel LBA: The cotransporter NaPi-IIb: characteristics, regulation and its role in carcinogenesis. Applied Cancer Res. 30:197–203. 2010.
11	Shibasaki Y, Etoh N and Hayasaka M: Targeted deletion of the tybe IIb Na⁺dependent Pi-co-transporter, NaPi-IIb, results in early embryonic lethality. Biochem Biophys Res Commun. 381:482–486. 2009.PubMed/NCBI
12	Kopantzev EP, Monastyrskaya GS, Vinogradova TV, et al: Differences in gene expression levels between early and later stages of human lung development are opposite to those between normal lung tissue and non-small lung cell carcinoma. Lung Cancer. 62:23–34. 2008. View Article : Google Scholar
13	el-Deiry WS, Nelkin BD, Celano P, Yen RW, Falco JP, Hamilton SR and Baylin SB: High expression of the DNA methyltransferase gene characterizes human neoplastic cells and progression stages of colon cancer. Proc Natl Acad Sci USA. 88:3470–3474. 1991. View Article : Google Scholar : PubMed/NCBI
14	Lengauer C, Kinzler KW and Vogelstein B: DNA methylation and genetic instability in colorectal cancer cells. Proc Natl Acad Sci USA. 94:2545–2550. 1997. View Article : Google Scholar : PubMed/NCBI
15	Liu H, Kho AT, Kohane IS and Sun Y: Predicting survival within the lung cancer histopathological hierarchy using a multi-scale genomic model of development. PLoS Med. 3:e2322006. View Article : Google Scholar
16	Ramirez MI, Pollack L, Millien G, Cao YX, Hinds A and Williams MC: The alpha-isoform of caveolin-1 is a marker of vasculogenesis in early lung development. J Histochem Cytochem. 50:33–42. 2002. View Article : Google Scholar : PubMed/NCBI
17	Hnasko R and Ben-Jonathan N: Developmental regulation of PV-1 in rat lung: association with the nuclear envelope and limited colocalization with Cav-1. Am J Physiol Lung Cell Mol Physiol. 288:L275–L284. 2005. View Article : Google Scholar
18	Wiliams TM and Lisanti MP: Caveolin-1 in oncogenic transformation, cancer, and metastasis. Am J Physiol Cell Physiol. 288:C494–C506. 2005. View Article : Google Scholar : PubMed/NCBI
19	Sunaga N, Miyajima K, Suzuki M, Sato M, White MA, Ramirez RD, Shay JW, Gazdar AF and Minna JD: Different roles for caveolin-1 in the development of non-small cell lung cancer versus small cell lung cancer. Cancer Res. 64:4277–4285. 2004. View Article : Google Scholar : PubMed/NCBI
20	Bélanger MM, Gaudreau M, Roussel E and Couet J: Role of caveolin-1 in etoposide resistance development in A549 lung cancer cells. Cancer Biol Ther. 3:954–959. 2004.PubMed/NCBI
21	Traebert M: Expression of type II Na-P(i) cotransporterin alveolar type II cells. Am J Physiol. 277:L868–L873. 1999.PubMed/NCBI
22	Grifﬁths MJ, Bonnet D and Janes SM: Stem cells of the alveolar epithelium. Lancet. 366:249–260. 2005.
23	Kinnard WV, Tuder R, Papst P and Fisher JH: Regulation of alveolar type II cell differentiation and proliferation in adult rat lung explants. Am J Respir Cell Mol Biol. 11:416–425. 1994. View Article : Google Scholar : PubMed/NCBI
24	Ten Have-Opbroek AA, Benfield JR, Hammond WG and Dijkman JH: Alveolar stem cells in canine bronchial carcinogenesis. Cancer Lett. 101:211–217. 1996.PubMed/NCBI
25	Ten Have-Opbroek AA, Benfield JR, van Krieken JH and Dijkman JH: The alveolar type II cell is a pluripotential stem cell in the genesis of human adenocarcinomas and squamous cell carcinomas. Histol Histopathol. 12:319–336. 1997.PubMed/NCBI
26	Kitinya JN, Sueishi K, Tanaka K and Katsuda Y: Immunoreactivity of surfactant-apoprotein adenocarcinomas, large cell and small cell carcinomas of the lung. Acta Pathol Jpn. 36:1271–1278. 1986.PubMed/NCBI
27	Gazdar FA, Linnoila R, Kurita Y, et al: Peripheral airway cell differentiation in human lung cancer. Cancer Res. 50:5481–5487. 1990.PubMed/NCBI
28	Holm BA, Matalon S, Finkelstein JN and Notter RH: Type II pneumocyte changes during hyperoxic lung injury and recovery. J Appl Physiol. 1985.65:2672–2678. 1988.PubMed/NCBI
29	Kim CF, Jackson EL, Woolfenden AE, et al: Identification of bronchioalveolar stem cells in normal lung and lung cancer. Cell. 121:823–835. 2005. View Article : Google Scholar : PubMed/NCBI
30	Xu CX, Jin H, Chung YS, et al: Low dietary inorganic phosphate affects the lung growth of developing mice. J Vet Sci. 10:105–113. 2009. View Article : Google Scholar : PubMed/NCBI
31	Tsao MS, Zhu H and Viallet J: Autocrine growth loop of the epidermal growth factor receptor in normal and immortalized human bronchial epithelial cells. Exp Cell Res. 223:268–273. 1996. View Article : Google Scholar : PubMed/NCBI
32	Lieber M, Smith B, Szakal A, Nelson-Rees W and Todaro G: A continuous tumor-cell line from a human lung carcinoma with properties of type II alveolar epithelial cells. Int J Cancer. 17:62–70. 1976. View Article : Google Scholar : PubMed/NCBI
33	Xu Q, Lu A, Xiao G, et al: Transcriptional profiling of midgut immunity response and degeneration in the wandering silkworm, Bombyx mori. Plos one. 7:e437692012. View Article : Google Scholar : PubMed/NCBI
34	Hill LD, Sun L, Leuschen MP and Zach TL: C3 synthesis by A549 alveolar epithelial cells is increased by interferon-γ and dexamethasone. Immunology. 79:236–240. 1993.PubMed/NCBI
35	Neal C, Neal M and Wickham H: Phosphate measurement in natural waters: two examples of analytical problems associated with silica interference using phosphomolybdic acid methodologies. Sci Total Environ. 252:511–512. 2000. View Article : Google Scholar
36	Gałeza M, Zebracka J, Szpak-Ulczok S, et al: Expression of selected genes involved in transport of ions in papillary thyroid carcinoma. Endokrynol Pol. 57:26–31. 2006.(In Polish).
37	Blanchard A, Shiu R, Booth S, et al: Gene expression profiling of early involuting mammary gland reveals novel genes potentially relevant to human breast cancer. Front Biosci. 12:2221–2232. 2007. View Article : Google Scholar
38	Yin BW, Kiyamova R, Chua R, Caballero OL, et al: Monoclonal antibody MX35 detects the membrane transporter NaPi2b (SLC34A2) in human carcinomas. Cancer Immun. 8:32008.PubMed/NCBI
39	Cuzin M: DNA chips: a new tool for genetic analysis and diagnostics. Transfus Clin Biol. 8:291–296. 2001. View Article : Google Scholar : PubMed/NCBI
40	Strunk RC, Eidlen DM and Mason RJ: Pulmonary alveolar type II epithelial cells synthesize and secrete proteins of the classical and alternative complement pathways. J Clin Invest. 81:1419–1426. 1988. View Article : Google Scholar : PubMed/NCBI
41	Niculescu F, Rus HG, Retegan M and Vlaicu R: Persistent complement activation on tumor cells in breast cancer. Am J Pathol. 140:1039–1043. 1992.PubMed/NCBI
42	Magyarlaki T, Mosolits S, Baranyay F and Buzogany I: Immunohistochemistry of complement response on human renal cell carcinoma biopsies. Tumori. 82:473–479. 1996.PubMed/NCBI
43	Clynes RA, Towers TL, Presta LG and Ravetch JV: Inhibitory Fc receptors modulate in vivo cytoxicity against tumor targets. Nat Med. 6:443–446. 2000. View Article : Google Scholar : PubMed/NCBI
44	Kierszenbaum F and Budzko DB: Cytotoxic effects of normal sera on lymphoid cells. I. Antibody-independent killing of heterologous thymocytes by guinea pig, rabbit, and human sera: Role of the alternative pathway of complement activation. Cell Immunol. 29:137–146. 1977.
45	Rothman BL, Despins AW and Kreutzer DL: Cytokine regulation of C3 and C5 production by the human type II pneumocyte cell line, A549. J Immunol. 145:592–598. 1990.PubMed/NCBI
46	Rothman BL, Merrow M, Despins A, Kennedy T and Kreutzer DL: Effect of lipopolysaccharide on C3 and C5 production by human lung cells. J Immunol. 143:196–202. 1989.PubMed/NCBI
47	Duerst RE, Ryan DH and Frantz CN: Variables affecting the killing of cultured human neuroblastoma cells with monoclonal antibody and complement. Cancer Res. 46:3420–3425. 1986.PubMed/NCBI
48	Jima DD, Shaha RN and Orcutta TM: Enhanced transcription of complement and coagulation genes in the absence of adaptive immunity. Mol Immunol. 46:1505–1516. 2009. View Article : Google Scholar : PubMed/NCBI
49	Hickman-Davis JM, Fang FC and Nathan C: Lung surfactant and reactive oxygen-nitrogen species: antimicrobial activity and host-pathogen interactions. Am J Physiol Lung Cell Mol Physiol. 281:L517–L523. 2001.PubMed/NCBI
50	Malmsten M, Lassen B, James MV and Nilsson UR: Adsorption of complement proteins C3 and C1q. J Colloid Interface Sci. 178:123–134. 1996. View Article : Google Scholar
51	Mold C: Effect of membrane phospholipids on activation of the alternative complement pathway. J Immunol. 143:1663–1668. 1989.PubMed/NCBI
52	Takeda E, Yamamoto H, Nashiki K, Sato T, Arai H and Taketani Y: Inorganic phosphate homeostasis and the role of dietary phosphorus. J Cell Mol Med. 8:191–200. 2004. View Article : Google Scholar : PubMed/NCBI
53	Furukawa T, Sunamura M, Motoi F, Matsuno S and Horii A: Potential tumor suppressive pathway involving DUSP6/MKP-3 in pancreatic cancer. Am J Pathol. 162:1807–1815. 2003. View Article : Google Scholar : PubMed/NCBI
54	Zhang Z, Kobayashi S, Borczuk AC, Leidner RS, Laframboise T, Levine AD and Halmos B: Dual speciﬁcity phosphatase 6 (DUSP6) is an ETS-regulated negative feedback mediator of oncogenic ERK signaling in lung cancer cells. Carcinogenesis. 31:577–586. 2010.
55	Li T, Yang W and Li M: Expression of selenium binding protein 1 characterizes intestinal cell maturation and predicts survival for patients with colorectal cancer. Mol Nutr Food Res. 52:1289–1299. 2008. View Article : Google Scholar : PubMed/NCBI
56	Huang KC, Park DC, Ng SK, et al: Selenium binding protein 1 in ovarian cancer. Int J Cancer. 118:2433–2440. 2006. View Article : Google Scholar : PubMed/NCBI
57	Chen G, Wang H and Miller CT: Reduced selenium-binding protein 1 expression is associated with poor outcome in lung adenocarcinomas. J Pathol. 202:321–329. 2004. View Article : Google Scholar : PubMed/NCBI
58	Yang M and Sytkowski AJ: Differential expression and androgen regulation of the human selenium-binding protein gene hSP56 in prostate cancer cells. Cancer Res. 58:3150–3153. 1998.PubMed/NCBI
59	Parikh H, Carlsson E, Chutkow WA and Johansson LE: TXNIP regulates peripheral glucose metabolism in humans. PLoS Med. 4:e1582007. View Article : Google Scholar : PubMed/NCBI
60	Capuano P, Bacic D and Stange G: Expression and regulation of the renal Na/phosphate cotransporter NaPi-IIa in a mouse model deficient for the PDZ protein PDZK1. Pflugers Arch. 449:392–402. 2005. View Article : Google Scholar : PubMed/NCBI