Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
Reproductive Biology and Endocrinology
Published in: Reproductive Biology and Endocrinology 1/2014

Open Access 01-12-2014 | Research

VEGF-D-induced draining lymphatic enlargement and tumor lymphangiogenesis promote lymph node metastasis in a xenograft model of ovarian carcinoma

Authors: Li-Cheng Du, Xian-Cheng Chen, Dong Wang, Yan-Jun Wen, Chun-Ting Wang, Xue-Mei Wang, Bing Kan, Yu-Quan Wei, Xia Zhao

Published in: Reproductive Biology and Endocrinology | Issue 1/2014

Login to get access

Abstract

Background

Vascular endothelial growth factor (VEGF)-D has been shown to promote lymph node metastasis in several cancers. Although generally overexpressed in ovarian carcinoma, its role in nodal dissemination of this cancer is unclear. To clarify the role of VEGF-D and the underlying molecular mechanisms, we investigated the function of VEGF-D using a mouse xenograft model of ovarian cancer.

Methods

Human ovarian serous adenocarcinoma SKOV3 cells were transfected with VEGF-D recombinant plasmid DNA, or with control vectors. The cells were injected subcutaneously into the footpads of nude mice. Tumor growth was evaluated weekly. Draining lymphatics were observed grossly with Evan’s blue lymphangiography. Tumoral lymphatics were delineated with both Evan’s blue and LYVE-1 immunostaining. Tumor metastases to lymph nodes were evaluated by H&E and CA125/CD40 staining. Expression of VEGF-D in primary tumors and levels of CA125 in involved lymph nodes were examined by immunohistochemistry. Tumor cell apoptosis was analyzed by Hoechst dyeing.

Results

Mice bearing VEGF-D overexpressing xenografts showed a significantly higher rate of lymph node metastasis and markedly greater tumor volume compared with the controls. The functional lymphatic vessels were denser and enlarged in marginal and central tumor portions. Additionally, higher CA125 expression was observed in the involved lymph nodes. Mice bearing VEGF-D overexpressing xenografts also exhibited a markedly lower apoptotic index compared with the controls.

Conclusions

Our data demonstrate the important role of VEGF-D in promoting lymph node metastasis by increasing tumor lymphangiogenesis, stimulating draining lymphatic vessel formation, and enhancing tumor invasiveness. Our findings show that VEGF-D can be a promising therapeutic target for ovarian cancer.
Appendix
Available only for authorised users
Literature
1.
di Re F, Baiocchi G, Fontanelli R, Grosso G, Cobellis L, Raspagliesi F, di Re E: Systematic pelvic and paraaortic lymphadenectomy for advanced ovarian cancer: prognostic significance of node metastases. Gynecol Oncol. 1996, 62 (3): 360-365. 10.1006/gyno.1996.0249.CrossRefPubMed
2.
Suzuki M, Ohwada M, Yamada T, Kohno T, Sekiguchi I, Sato I: Lymph node metastasis in stage I epithelial ovarian cancer. Gynecol Oncol. 2000, 79 (2): 305-308. 10.1006/gyno.2000.5951.CrossRefPubMed
3.
Li L, Liu B, Li X, Yang S, Xiao J, Chen M, Zhang Y, Ma J: Vascular endothelial growth factor D and intratumoral lymphatics as independent prognostic factors in epithelial ovarian carcinoma. Anat Rec (Hoboken). 2009, 292 (4): 562-569. 10.1002/ar.20845.CrossRef
4.
Yokoyama Y, Charnock-Jones DS, Licence D, Yanaihara A, Hastings JM, Holland CM, Emoto M, Umemoto M, Sakamoto T, Sato S, Mizunuma H, Smith SK: Vascular endothelial growth factor-D is an independent prognostic factor in epithelial ovarian carcinoma. Br J Cancer. 2003, 88 (2): 237-244. 10.1038/sj.bjc.6600701.PubMedCentralCrossRefPubMed
5.
Morice P, Joulie F, Camatte S, Atallah D, Rouzier R, Pautier P, Pomel C, Lhommé C, Duvillard P, Castaigne D: Lymph node involvement in epithelial ovarian cancer: analysis of 276 pelvic and paraaortic lymphadenectomies and surgical implications. J Am Coll Surg. 2003, 197 (2): 198-205. 10.1016/S1072-7515(03)00234-5.CrossRefPubMed
6.
7.
de Poncheville L, Perrotin F, Lefrancq T, Lansac J, Body G: Does paraaortic lymphadenectomy have a benefit in the treatment of ovarian cancer that is apparently confined to the ovaries?. Eur J Cancer. 2001, 37 (2): 210-215. 10.1016/S0959-8049(00)00377-4.CrossRefPubMed
8.
Shirai Y, Wakai T, Hatakeyama K: Radical lymph node dissection for gallbladder cancer: indications and limitations. Surg Oncol Clin N Am. 2007, 16 (1): 221-232. 10.1016/j.soc.2006.10.011.CrossRefPubMed
9.
Stacker SA, Stenvers K, Caesar C, Vitali A, Domagala T, Nice E, Roufail S, Simpson RJ, Moritz R, Karpanen T, Alitalo K, Achen MG: Biosynthesis of vascular endothelial growth factor-D involves proteolytic processing which generates non-covalent homodimers. J Biol Chem. 1999, 274 (45): 32127-32136. 10.1074/jbc.274.45.32127.CrossRefPubMed
10.
Harris NC, Paavonen K, Davydova N, Roufail S, Sato T, Zhang YF, Karnezis T, Stacker SA, Achen MG: Proteolytic processing of vascular endothelial growth factor-D is essential for its capacity to promote the growth and spread of cancer. FASEB J. 2011, 25 (8): 2615-2625. 10.1096/fj.10-179788.CrossRefPubMed
11.
Kärpänen T, Heckman CA, Keskitalo S, Jeltsch M, Ollila H, Neufeld G, Tamagnone L, Alitalo K: Functional interaction of VEGF-C and VEGF-D with neuropilin receptors. FASEB J. 2006, 20 (9): 1462-1472. 10.1096/fj.05-5646com.CrossRefPubMed
12.
Thelen A, Scholz A, Benckert C, von Marschall Z, Schröder M, Wiedenmann B, Neuhaus P, Rosewicz S, Jonas S: VEGF-D promotes tumor growth and lymphatic spread in a mouse model of hepatocellular carcinoma. Int J Cancer. 2008, 122 (11): 2471-2481. 10.1002/ijc.23439.CrossRefPubMed
13.
Lin W, Jiang L, Chen Y, She F, Han S, Zhu J, Zhou L, Tang N, Wang X, Li X: Vascular endothelial growth factor-D promotes growth, lymphangiogenesis and lymphatic metastasis in gallbladder cancer. Cancer Lett. 2012, 314 (2): 127-136. 10.1016/j.canlet.2011.09.004.CrossRefPubMed
14.
Kozlowski M, Naumnik W, Niklinski J, Milewski R, Dziegielewski P, Laudanski J: Vascular endothelial growth factor C and D expression correlates with lymph node metastasis and poor prognosis in patients with resected esophageal cancer. Neoplasma. 2011, 58 (4): 311-319. 10.4149/neo_2011_04_311.CrossRefPubMed
15.
Herrmann E, Eltze E, Bierer S, Kopke T, Gorge T, Neumann J, Hertle L, Wulfing C: VEGF-C, VEGF-D and Flt-4 in transitional bladder cancer: relationships to clinicopathological parameters and long-term survival. Anticancer Res. 2007, 27 (5A): 3127-3133.PubMed
16.
Niki T, Iba S, Tokunou M, Yamada T, Matsuno Y, Hirohashi S: Expression of vascular endothelial growth factors A, B, C, and D and their relationships to lymph node status in lung adenocarcinoma. Clin Cancer Res. 2000, 6 (6): 2431-2439.PubMed
17.
Girling JE, Donoghue JF, Lederman FL, Cann LM, Achen MG, Stacker SA, Rogers PA: Vascular endothelial growth factor-D over-expressing tumor cells induce differential effects on uterine vasculature in a mouse model of endometrial cancer. Reprod Biol Endocrinol. 2010, 8: 84-10.1186/1477-7827-8-84.PubMedCentralCrossRefPubMed
18.
Mohammed RA, Green A, El-Shikh S, Paish EC, Ellis IO, Martin SG: Prognostic significance of vascular endothelial cell growth factors -A, -C and -D in breast cancer and their relationship with angio- and lymphangiogenesis. Br J Cancer. 2007, 96 (7): 1092-1100. 10.1038/sj.bjc.6603678.PubMedCentralCrossRefPubMed
19.
Thériault C, Pinard M, Comamala M, Migneault M, Beaudin J, Matte I, Boivin M, Piché A, Rancourt C: MUC16 (CA125) regulates epithelial ovarian cancer cell growth, tumorigenesis and metastasis. Gynecol Oncol. 2011, 121 (3): 434-443. 10.1016/j.ygyno.2011.02.020.CrossRefPubMed
20.
Hakkarainen T, Hemminki A, Pereboev A, Barker S, Asiedu C, Strong T, Kanerva A, Wahlfors J, Curiel D: CD40 is expressed on ovarian cancer cells and can be utilized for targeting adenoviruses. Clin Cancer Res. 2003, 9 (2): 619-624.PubMed
21.
Gallagher NJ, Eliopoulos AG, Agathangelo A, Oates J, Crocker J, Young LS: CD40 activation in epithelial ovarian carcinoma cells modulates growth, apoptosis, and cytokine secretion. Mol Pathol. 2002, 55 (2): 110-120. 10.1136/mp.55.2.110.PubMedCentralCrossRefPubMed
22.
Wissmann C, Detmar M: Pathways targeting tumor lymphangiogenesis. Clin Cancer Res. 2006, 12 (23): 6865-6868. 10.1158/1078-0432.CCR-06-1800.CrossRefPubMed
23.
Tobler NE, Detmar M: Tumor and lymph node lymphangiogenesis–impact on cancer metastasis. J Leukoc Biol. 2006, 80 (4): 691-696. 10.1189/jlb.1105653.CrossRefPubMed
24.
25.
Skobe M, Hawighorst T, Jackson DG, Prevo R, Janes L, Velasco P, Riccardi L, Alitalo K, Claffey K, Detmar M: Induction of tumor lymphangiogenesis by VEGF-C promotes breast cancer metastasis. Nat Med. 2001, 7 (2): 192-198. 10.1038/84643.CrossRefPubMed
26.
Mandriota SJ, Jussila L, Jeltsch M, Compagni A, Baetens D, Prevo R, Banerji S, Huarte J, Montesano R, Jackson DG, Orci L, Alitalo K, Christofori G, Pepper MS: Vascular endothelial growth factor-C-mediated lymphangiogenesis promotes tumour metastasis. EMBO J. 2001, 20 (4): 672-682. 10.1093/emboj/20.4.672.PubMedCentralCrossRefPubMed
27.
Chaudary N, Milosevic M, Hill RP: Suppression of vascular endothelial growth factor receptor 3 (VEGFR3) and vascular endothelial growth factor C (VEGFC) inhibits hypoxia-induced lymph node metastases in cervix cancer. Gynecol Oncol. 2011, 123 (2): 393-400. 10.1016/j.ygyno.2011.07.006.CrossRefPubMed
28.
Shimizu K, Kubo H, Yamaguchi K, Kawashima K, Ueda Y, Matsuo K, Awane M, Shimahara Y, Takabayashi A, Yamaoka Y, Satoh S: Suppression of VEGFR-3 signaling inhibits lymph node metastasis in gastric cancer. Cancer Sci. 2004, 95 (4): 328-333. 10.1111/j.1349-7006.2004.tb03211.x.CrossRefPubMed
29.
Yasuoka H, Nakamura Y, Zuo H, Tang W, Takamura Y, Miyauchi A, Nakamura M, Mori I, Kakudo K: VEGF-D expression and lymph vessels play an important role for lymph node metastasis in papillary thyroid carcinoma. Mod Pathol. 2005, 18 (8): 1127-1133. 10.1038/modpathol.3800402.CrossRefPubMed
30.
Kurahara H, Takao S, Maemura K, Shinchi H, Natsugoe S, Aikou T: Impact of vascular endothelial growth factor-C and -D expression in human pancreatic cancer: its relationship to lymph node metastasis. Clin Cancer Res. 2004, 10 (24): 8413-8420. 10.1158/1078-0432.CCR-04-0379.CrossRefPubMed
31.
Nakamura Y, Yasuoka H, Tsujimoto M, Yang Q, Imabun S, Nakahara M, Nakao K, Nakamura M, Mori I, Kakudo K: Prognostic significance of vascular endothelial growth factor D in breast carcinoma with long-term follow-up. Clin Cancer Res. 2003, 9 (2): 716-721.PubMed
32.
Gisterek I, Matkowski R, Kozlak J, Dus D, Lacko A, Szelachowska J, Kornafel J: Evaluation of prognostic value of VEGF-C and VEGF-D in breast cancer–10 years follow-up analysis. Anticancer Res. 2007, 27 (4C): 2797-2802.PubMed
33.
Zhao YC, Ni XJ, Wang MH, Zha XM, Zhao Y, Wang S: Tumor-derived VEGF-C, but not VEGF-D, promotes sentinel lymph node lymphangiogenesis prior to metastasis in breast cancer patients. Med Oncol. 2012, 29 (4): 2594-2600. 10.1007/s12032-012-0205-0.CrossRefPubMed
34.
Rossochacka-Rostalska B, Gisterek I, Suder E, Matkowski R, Szelachowska J, Kornafel J: Vascular endothelial growth factor-C (VEGF-C) and vascular endothelial growth factor-D (VEGF-D) in ovarian carcinomas. Ginekol Pol. 2006, 77 (11): 830-839.PubMed
35.
Massi D, Puig S, Franchi A, Malvehy J, Vidal-Sicart S, González-Cao M, Baroni G, Ketabchi S, Palou J, Santucci M: Tumour lymphangiogenesis is a possible predictor of sentinel lymph node status in cutaneous melanoma: a case–control study. J Clin Pathol. 2006, 59 (2): 166-173. 10.1136/jcp.2005.028431.PubMedCentralCrossRefPubMed
36.
Hirakawa S, Brown LF, Kodama S, Paavonen K, Alitalo K, Detmar M: VEGF-C-induced lymphangiogenesis in sentinel lymph nodes promotes tumor metastasis to distant sites. Blood. 2007, 109 (3): 1010-1017.PubMedCentralCrossRefPubMed
37.
38.
39.
Kaipainen A, Korhonen J, Mustonen T, van Hinsbergh VW, Fang GH, Dumont D, Breitman M, Alitalo K: Expression of the fms-like tyrosine kinase 4 gene becomes restricted to lymphatic endothelium during development. Proc Natl Acad Sci U S A. 1995, 92 (8): 3566-3570. 10.1073/pnas.92.8.3566.PubMedCentralCrossRefPubMed
40.
Fitz LJ, Morris JC, Towler P, Long A, Burgess P, Greco R, Wang J, Gassaway R, Nickbarg E, Kovacic S, Ciarletta A, Giannotti J, Finnerty H, Zollner R, Beier DR, Leak LV, Turner KJ, Wood CR: Characterization of murine Flt4 ligand/VEGF-C. Oncogene. 1997, 15 (5): 613-618. 10.1038/sj.onc.1201191.CrossRefPubMed
41.
Karnezis T, Shayan R, Caesar C, Roufail S, Harris NC, Ardipradja K, Zhang YF, Williams SP, Farnsworth RH, Chai MG, Rupasinghe TW, Tull DL, Baldwin ME, Sloan EK, Fox SB, Achen MG, Stacker SA: VEGF-D promotes tumor metastasis by regulating prostaglandins produced by the collecting lymphatic endothelium. Cancer Cell. 2012, 21 (2): 181-195. 10.1016/j.ccr.2011.12.026.CrossRefPubMed
42.
Yu M, Berk R, Kosir MA: CXCL7-Mediated stimulation of lymphangiogenic factors VEGF-C, VEGF-D in human breast cancer cells. J Oncol. 2010, 2010: 939407-PubMedCentralCrossRefPubMed
43.
Yasuoka H, Kodama R, Hirokawa M, Takamura Y, Miyauchi A, Inagaki M, Sanke T, Nakamura Y: Neuropilin-2 expression in papillary thyroid carcinoma: correlation with VEGF-D expression, lymph node metastasis, and VEGF-D-induced aggressive cancer cell phenotype. J Clin Endocrinol Metab. 2011, 96 (11): E1857-E1861. 10.1210/jc.2011-1180.CrossRefPubMed
44.
Reinartz S, Failer S, Schuell T, Wagner U: CA125 (MUC16) gene silencing suppresses growth properties of ovarian and breast cancer cells. Eur J Cancer. 2012, 48 (10): 1558-1569. 10.1016/j.ejca.2011.07.004.CrossRefPubMed
45.
Nagamoto N, Saito Y, Ohta S, Sato M, Kanma K, Sagawa M, Takahashi S, Usuda K, Nakada T, Hashimoto K: Relationship of lymph node metastasis to primary tumor size and microscopic appearance of roentgenographically occult lung cancer. Am J Surg Pathol. 1989, 13 (12): 1009-1013. 10.1097/00000478-198912000-00002.CrossRefPubMed
46.
Carter CL, Allen C, Henson DE: Relation of tumor size, lymph node status, and survival in 24,740 breast cancer cases. Cancer. 1989, 63 (1): 181-187. 10.1002/1097-0142(19890101)63:1<181::AID-CNCR2820630129>3.0.CO;2-H.CrossRefPubMed
47.
Nathanson SD, Anaya P, Avery M, Hetzel FW, Sarantou T, Havstad S: Sentinel lymph node metastasis in experimental melanoma: relationships among primary tumor size, lymphatic vessel diameter and 99mTc-labeled human serum albumin clearance. Ann Surg Oncol. 1997, 4 (2): 161-168. 10.1007/BF02303800.CrossRefPubMed
48.
Chung EJ, Rho YS, Baek SK, Woo JS, Kwon SY, Lee NJ, Chae YS, Jung KY: Does a growing tumour volume induce lymphangiogenesis? A study of oral/oropharyngeal cancer. J Otolaryngol Head Neck Surg. 2011, 40 (4): 311-317.PubMed
49.
Akahane M, Akahane T, Matheny SL, Shah A, Okajima E, Thorgeirsson UP: Vascular endothelial growth factor-D is a survival factor for human breast carcinoma cells. Int J Cancer. 2006, 118 (4): 841-849. 10.1002/ijc.21420.CrossRefPubMed
50.
Valtola R, Salven P, Heikkila P, Taipale J, Joensuu H, Rehn M, Pihlajaniemi T, Weich H, de Waal R, Alitalo K: VEGFR-3 and its ligand VEGF-C are associated with angiogenesis in breast cancer. Am J Pathol. 1999, 154 (5): 1381-1390. 10.1016/S0002-9440(10)65392-8.PubMedCentralCrossRefPubMed
51.
Stacker SA, Caesar C, Baldwin ME, Thornton GE, Williams RA, Prevo R, Jackson DG, Nishikawa S, Kubo H, Achen MG: VEGF-D promotes the metastatic spread of tumor cells via the lymphatics. Nat Med. 2001, 7 (2): 186-191. 10.1038/84635.CrossRefPubMed
Metadata
Title
VEGF-D-induced draining lymphatic enlargement and tumor lymphangiogenesis promote lymph node metastasis in a xenograft model of ovarian carcinoma
Authors
Li-Cheng Du
Xian-Cheng Chen
Dong Wang
Yan-Jun Wen
Chun-Ting Wang
Xue-Mei Wang
Bing Kan
Yu-Quan Wei
Xia Zhao
Publication date
01-12-2014
Publisher
BioMed Central
Published in
Reproductive Biology and Endocrinology / Issue 1/2014
Electronic ISSN: 1477-7827
DOI
https://doi.org/10.1186/1477-7827-12-14

Other articles of this Issue 1/2014

Reproductive Biology and Endocrinology 1/2014 Go to the issue

Research

Delaying the oocyte maturation trigger by one day leads to a higher metaphase II oocyte yield in IVF/ICSI: a randomised controlled trial

Review

Making gametes from pluripotent stem cells – a promising role for very small embryonic-like stem cells

Research

The thrombospondin-1 receptor CD36 is an important mediator of ovarian angiogenesis and folliculogenesis

Research

Fertility awareness among medical and non-medical students: a case-control study

Research

Alteration of Th17 and Treg cells in patients with unexplained recurrent spontaneous abortion before and after lymphocyte immunization therapy

Research

The prevalence of metabolic disorders in various phenotypes of polycystic ovary syndrome: a community based study in Southwest of Iran