Recombinant HE4 protein promotes proliferation of pancreatic and endometrial cancer cell lines

Lu,Qinsheng; Chen,Haibin; Senkowski,Christopher; Wang,Jianhao; Wang,Xue; Brower,Steven; Glasgow,Wayne; Byck,David; Jiang,Shi-Wen; Li,Jinping

doi:10.3892/or.2015.4339

Recombinant HE4 protein promotes proliferation of pancreatic and endometrial cancer cell lines

Authors:
- Qinsheng Lu
- Haibin Chen
- Christopher Senkowski
- Jianhao Wang
- Xue Wang
- Steven Brower
- Wayne Glasgow
- David Byck
- Shi-Wen Jiang
- Jinping Li
View Affiliations

Affiliations: Department of Histology and Embryology, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China, Curtis and Elizabeth Anderson Cancer Institute, Department of Surgery, Memorial Health University Medical Center, Savannah, GA 31404, USA, School of Pharmaceutical Engineering and Life Science, Changzhou University, Changzhou, Jiangsu 213164, P.R. China, Department of Biomedical Science, Mercer University School of Medicine, Savannah, GA 31404, USA, Department of Surgery and Surgical Oncology, Beth Israel Medical Center, New York, NY 10003, USA, Department of Obstetrics and Gynecology, Memorial Health University Medical Center, Savannah, GA 31404, USA
Published online on: October 20, 2015 https://doi.org/10.3892/or.2015.4339
Pages: 163-170

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Pancreatic adenocarcinoma is one of the most deadly malignancies, and endometrial cancer represents the most common gynecologic cancer in the USA. Better understanding on the pathologic mechanisms and pathways is required for effective treatment of these malignancies. Recently, human epididymis protein 4 (HE4 or WFDC2), a secretory glycoprotein, was found to be overexpressed in pancreatic and endometrial cancers. In addition, studies have shown that HE4 overexpression in endometrial cancer cell lines led to faster cancer progression in a mouse subcutaneous model. These findings raise a question on the role(s) of secretory, extracellular HE4 in cancer development. In the present study, we found that treatment of pancreatic and endometrial cancer cell lines with purified, extracellular HE4 protein led to a significant increase in cell viability and proliferation. Moreover, extracellular HE4 protein was able to increase DNA synthesis, and modulate the mRNA and protein levels of cell cycle marker PCNA and cell cycle inhibitor p21. These effects appeared to be robust and sustainable and required a relatively low concentration of HE4 protein. The findings indicated the secreted, extracellular HE4 may carry some physiopathological functions. Via paracrine/endocrine actions, circulatory HE4 produced by malignant cells may contribute to pancreatic and endometrial cancer progression and/or metastasis.

View Figures

View References

1	Siegel R, Ma J, Zou Z and Jemal A: Cancer statistics, 2014. CA Cancer J Clin. 64:9–29. 2014. View Article : Google Scholar : PubMed/NCBI
2	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar : PubMed/NCBI
3	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 127:2893–2917. 2010. View Article : Google Scholar
4	Bardeesy N and DePinho RA: Pancreatic cancer biology and genetics. Nat Rev Cancer. 2:897–909. 2002. View Article : Google Scholar : PubMed/NCBI
5	Muniraj T, Jamidar PA and Aslanian HR: Pancreatic cancer: A comprehensive review and update. Dis Mon. 59:368–402. 2013. View Article : Google Scholar : PubMed/NCBI
6	Sakai T, Toguchida J, Ohtani N, Yandell DW, Rapaport JM and Dryja TP: Allele-specific hypermethylation of the retinoblastoma tumor-suppressor gene. Am J Hum Genet. 48:880–888. 1991.PubMed/NCBI
7	Esteller M: CpG island hypermethylation and tumor suppressor genes: A booming present, a brighter future. Oncogene. 21:5427–5440. 2002. View Article : Google Scholar : PubMed/NCBI
8	Kirchhoff C, Habben I, Ivell R and Krull N: A major human epididymis-specific cDNA encodes a protein with sequence homology to extracellular proteinase inhibitors. Biol Reprod. 45:350–357. 1991. View Article : Google Scholar : PubMed/NCBI
9	Clauss A, Lilja H and Lundwall A: A locus on human chromosome 20 contains several genes expressing protease inhibitor domains with homology to whey acidic protein. Biochem J. 368:233–242. 2002. View Article : Google Scholar : PubMed/NCBI
10	Bingle CD: Towards defining the complement of mammalian WFDC-domain-containing proteins. Biochem Soc Trans. 39:1393–1397. 2011. View Article : Google Scholar : PubMed/NCBI
11	Galgano MT, Hampton GM and Frierson HF Jr: Comprehensive analysis of HE4 expression in normal and malignant human tissues. Mod Pathol. 19:847–853. 2006.PubMed/NCBI
12	Jiang SW, Chen H, Dowdy S, Fu A, Attewell J, Kalogera E, Drapkin R, Podratz K, Broaddus R and Li J: HE4 transcription- and splice variants-specific expression in endometrial cancer and correlation with patient survival. Int J Mol Sci. 14:22655–22677. 2013. View Article : Google Scholar : PubMed/NCBI
13	Drapkin R, von Horsten HH, Lin Y, Mok SC, Crum CP, Welch WR and Hecht JL: Human epididymis protein 4 (HE4) is a secreted glycoprotein that is overexpressed by serous and endometrioid ovarian carcinomas. Cancer Res. 65:2162–2169. 2005. View Article : Google Scholar : PubMed/NCBI
14	Ryu B, Jones J, Blades NJ, Parmigiani G, Hollingsworth MA, Hruban RH and Kern SE: Relationships and differentially expressed genes among pancreatic cancers examined by large-scale serial analysis of gene expression. Cancer Res. 62:819–826. 2002.PubMed/NCBI
15	Huang T, Jiang SW, Qin L, Senkowski C, Lyle C, Terry K, Brower S, Chen H, Glasgow W, Wei Y, et al: Expression and diagnostic value of HE4 in pancreatic adenocarcinoma. Int J Mol Sci. 16:2956–2970. 2015. View Article : Google Scholar : PubMed/NCBI
16	O'Neal RL, Nam KT, LaFleur BJ, Barlow B, Nozaki K, Lee HJ, Kim WH, Yang HK, Shi C, Maitra A, et al: Human epididymis protein 4 is up-regulated in gastric and pancreatic adenocarcinomas. Hum Pathol. 44:734–742. 2013. View Article : Google Scholar
17	Angioli R, Miranda A, Aloisi A, Montera R, Capriglione S, De Cicco Nardone C, Terranova C and Plotti F: A critical review on HE4 performance in endometrial cancer: Where are we now? Tumour Biol. 35:881–887. 2014. View Article : Google Scholar
18	Moore RG, Miller CM, Brown AK, Robison K, Steinhoff M and Lambert-Messerlian G: Utility of tumor marker HE4 to predict depth of myometrial invasion in endometrioid adenocarcinoma of the uterus. Int J Gynecol Cancer. 21:1185–1190. 2011.PubMed/NCBI
19	Faca VM, Song KS, Wang H, Zhang Q, Krasnoselsky AL, Newcomb LF, Plentz RR, Gurumurthy S, Redston MS, Pitteri SJ, et al: A mouse to human search for plasma proteome changes associated with pancreatic tumor development. PLoS Med. 5:e1232008. View Article : Google Scholar : PubMed/NCBI
20	Brand RE, Nolen BM, Zeh HJ, Allen PJ, Eloubeidi MA, Goldberg M, Elton E, Arnoletti JP, Christein JD, Vickers SM, et al: Serum biomarker panels for the detection of pancreatic cancer. Clin Cancer Res. 17:805–816. 2011. View Article : Google Scholar : PubMed/NCBI
21	Bingle CD and Vyakarnam A: Novel innate immune functions of the whey acidic protein family. Trends Immunol. 29:444–453. 2008. View Article : Google Scholar : PubMed/NCBI
22	Scott A, Weldon S and Taggart CC: SLPI and elafin: Multifunctional antiproteases of the WFDC family. Biochem Soc Trans. 39:1437–1440. 2011. View Article : Google Scholar : PubMed/NCBI
23	Chhikara N, Saraswat M, Tomar AK, Dey S, Singh S and Yadav S: Human epididymis protein-4 (HE-4): A novel cross-class protease inhibitor. PLoS One. 7:e476722012. View Article : Google Scholar : PubMed/NCBI
24	LeBleu VS, Teng Y, O'Connell JT, Charytan D, Müller GA, Müller CA, Sugimoto H and Kalluri R: Identification of human epididymis protein-4 as a fibroblast-derived mediator of fibrosis. Nat Med. 19:227–231. 2013. View Article : Google Scholar : PubMed/NCBI
25	Hua L, Liu Y, Zhen S, Wan D, Cao J and Gao X: Expression and biochemical characterization of recombinant human epididymis protein 4. Protein Expr Purif. 102:52–62. 2014. View Article : Google Scholar : PubMed/NCBI
26	Bignotti E, Ragnoli M, Zanotti L, Calza S, Falchetti M, Lonardi S, Bergamelli S, Bandiera E, Tassi RA, Romani C, et al: Diagnostic and prognostic impact of serum HE4 detection in endometrial carcinoma patients. Br J Cancer. 104:1418–1425. 2011. View Article : Google Scholar : PubMed/NCBI
27	Kalogera E, Scholler N, Powless C, Weaver A, Drapkin R, Li J, Jiang SW, Podratz K, Urban N and Dowdy SC: Correlation of serum HE4 with tumor size and myometrial invasion in endometrial cancer. Gynecol Oncol. 124:270–275. 2012. View Article : Google Scholar
28	Zanotti L, Bignotti E, Calza S, Bandiera E, Ruggeri G, Galli C, Tognon G, Ragnoli M, Romani C, Tassi RA, et al: Human epididymis protein 4 as a serum marker for diagnosis of endometrial carcinoma and prediction of clinical outcome. Clin Chem Lab Med. 50:2189–2198. 2012. View Article : Google Scholar : PubMed/NCBI
29	Angioli R, Plotti F, Capriglione S, Montera R, Damiani P, Ricciardi R, Aloisi A, Luvero D, Cafà EV, Dugo N, et al: The role of novel biomarker HE4 in endometrial cancer: A case control prospective study. Tumour Biol. 34:571–576. 2013. View Article : Google Scholar
30	Lu R, Sun X, Xiao R, Zhou L and Gaoxand Guo L: Human epididymis protein 4 (HE4) plays a key role in ovarian cancer cell adhesion and motility. Biochem Biophys Res Commun. 419:274–280. 2012. View Article : Google Scholar : PubMed/NCBI
31	Li J, Chen H, Mariani A, Chen D, Klatt E, Podratz K, Drapkin R, Broaddus R, Dowdy S and Jiang SW: HE4 (WFDC2) promotes tumor growth in endometrial cancer cell lines. Int J Mol Sci. 14:6026–6043. 2013. View Article : Google Scholar : PubMed/NCBI
32	Zhu YF, Gao GL, Tang SB, Zhang ZD and Huang QS: Effect of WFDC 2 silencing on the proliferation, motility and invasion of human serous ovarian cancer cells in vitro. Asian Pac J Trop Med. 6:265–272. 2013. View Article : Google Scholar : PubMed/NCBI
33	Chen Y, Mu X, Wang S, Zhao L, Wu Y, Li J and Li M: WAP four-disulfide core domain protein 2 mediates the proliferation of human ovarian cancer cells through the regulation of growth-and apoptosis-associated genes. Oncol Rep. 29:288–296. 2013.
34	Wang H, Zhu L, Gao J, Hu Z and Lin B: Promotive role of recombinant HE4 protein in proliferation and carboplatin resistance in ovarian cancer cells. Oncol Rep. 33:403–412. 2015.
35	Schmittgen TD and Livak KJ: Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 3:1101–1108. 2008. View Article : Google Scholar : PubMed/NCBI
36	Ahrendt SA and Pitt HA: Surgical management of pancreatic cancer. Oncology. 16:725–743. 2002.PubMed/NCBI
37	Missiaglia E, Blaveri E, Terris B, Wang YH, Costello E, Neoptolemos JP, Crnogorac-Jurcevic T and Lemoine NR: Analysis of gene expression in cancer cell lines identifies candidate markers for pancreatic tumorigenesis and metastasis. Int J Cancer. 112:100–112. 2004. View Article : Google Scholar : PubMed/NCBI
38	Hamilton CA, Cheung MK, Osann K, Chen L, Teng NN, Longacre TA, Powell MA, Hendrickson MR, Kapp DS and Chan JK: Uterine papillary serous and clear cell carcinomas predict for poorer survival compared to grade 3 endometrioid corpus cancers. Br J Cancer. 94:642–646. 2006.PubMed/NCBI
39	Zou SL, Chang XH, Ye X, Cheng HY, Cheng YX, Tang ZJ, Zhang ZJ, Gao L, Chen XH and Cui H: Effect of human epididymis protein 4 gene silencing on the malignant phenotype in ovarian cancer. Chin Med J. 124:3133–3140. 2011.PubMed/NCBI
40	Moore RG, Hill EK, Horan T, Yano N, Kim K, MacLaughlan S, Lambert-Messerlian G, Tseng YD, Padbury JF, Miller MC, et al: HE4 (WFDC2) gene overexpression promotes ovarian tumor growth. Sci Rep. 4:35742014. View Article : Google Scholar : PubMed/NCBI
41	Kelman Z: PCNA: Structure, functions and interactions. Oncogene. 14:629–640. 1997. View Article : Google Scholar : PubMed/NCBI
42	Moldovan GL, Pfander B and Jentsch S: PCNA controls establishment of sister chromatid cohesion during S phase. Mol Cell. 23:723–732. 2006. View Article : Google Scholar : PubMed/NCBI
43	Harper JW, Adami GR, Wei N, Keyomarsi K and Elledge SJ: The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. Cell. 75:805–816. 1993. View Article : Google Scholar : PubMed/NCBI
44	Xiong Y, Hannon GJ, Zhang H, Casso D, Kobayashi R and Beach D: p21 is a universal inhibitor of cyclin kinases. Nature. 366:701–704. 1993. View Article : Google Scholar : PubMed/NCBI
45	Waga S, Hannon GJ, Beach D and Stillman B: The p21 inhibitor of cyclin-dependent kinases controls DNA replication by interaction with PCNA. Nature. 369:574–578. 1994. View Article : Google Scholar : PubMed/NCBI
46	Chen J, Jackson PK, Kirschner MW and Dutta A: Separate domains of p21 involved in the inhibition of Cdk kinase and PCNA. Nature. 374:386–388. 1995. View Article : Google Scholar : PubMed/NCBI