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[Cancer Research 60, 4324-4327, August 15, 2000]
© 2000 American Association for Cancer Research

Advances in Brief

Absence of Functional Lymphatics within a Murine Sarcoma: A Molecular and Functional Evaluation1

Anders J. Leu2, David A. Berk3, Athina Lymboussaki, Kari Alitalo and Rakesh K. Jain4

Edwin L. Steele Laboratory, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114 [A. J. L., D. A. B., R. K. J.], and Molecular/Cancer Biology Laboratory and Ludwig Institute for Cancer Research, Haartman Institute, University of Helsinki, SF-00014 Helsinki, Finland [A. L., K. A.]


   ABSTRACT
Top
 ABSTRACT
Introduction
 Materials and Methods
 Results and Discussion
 REFERENCES
 
Despite a clinically recognized association between the lymphatics and metastasis, the biology of tumor-lymphatic interaction is not clearly understood. We report here that functional lymphatic capillaries are absent from the interior of a solid tumor, despite the presence within the tumor of the lymphangiogenic molecule vascular endothelial growth factor (VEGF)-C and endothelial cells bearing its receptor, VEGF receptor 3. Functional lymphatics, enlarged and VEGF receptor 3 positive, were detected in some tumors only at the tumor periphery (within 100 µm of the interface with normal tissue). We conclude that although lymphangiogenic factors are present, formation of functional lymphatic vessels is prevented, possibly due to collapse by the solid stress exerted by growing cancer cells.


   Introduction
Top
 ABSTRACT
 Introduction
Materials and Methods
 Results and Discussion
 REFERENCES
 
Tumor-induced lymphangiogenesis might be a factor in metastatic progression; however, to date, this speculation is not supported by direct evidence (1 , 2) . Most evidence indicates that lymphatics are absent from tumors (3, 4, 5, 6, 7) , although the unambiguous detection of tumor lymphatics by histological methods has been problematic due to the poorly differentiated vascular morphology and questions of marker specificity (8) . A group of endothelial-associated receptor tyrosine kinases (VEGFR-1,5 VEGFR-2, and VEGFR-3) may be useful for the study of tumor-lymphatic interaction. Specifically, VEGFR-3 may be an important molecule for lymphangiogenesis because the ligand for this receptor, VEGF-C (9 , 10) , was demonstrated in vivo to be a potent mitogen for lymphatic vessels (11) . VEGFR-3 and VEGF-C have also been shown to be expressed in a variety of human tumors (12, 13, 14, 15) and have been correlated with mortality (16) .

Here we describe a combined molecular and functional characterization of endothelial cells in a murine sarcoma, FSaII, implanted in the tail skin of nude mice. To assess the functional status of the lymphatic endothelial cells, we performed in vivo fluorescence and ferritin microlymphography (17, 18, 19, 20) to mark lymph and detect superficial and deeper lymphatic capillaries capable of forming and transporting lymph. We used in situ hybridization to examine the distribution of VEGF-C, VEGFR-3, and the general endothelial marker VEGFR-2 (Flk1) in the tumor allograft and normal dermis.


   Materials and Methods
Top
 ABSTRACT
 Introduction
 Materials and Methods
Results and Discussion
 REFERENCES
 
Animals and Tumor Model.
Functional and histological studies were performed on 20 nude mice (female; age, 29 days; weight, 17.8 ± 2.2 g). Tumors were also implanted in six additional mice for the in situ hybridization study. All procedures were performed in accordance with Massachusetts General Hospital animal welfare guidelines.

Functional and Histological Studies.
FITC-dextran (Mr 2,000,000; Sigma Chemical Co., St. Louis, MO) was used as a lymph marker for in vivo fluorescence imaging (17, 18, 19, 20) , and ferritin (type I ferritin from horse spleen; Mr 480,000; Sigma Chemical Co.) was injected as the lymph marker for subsequent histochemical staining. On day 1 of the study, fluorescence microlymphography was performed to delineate the superficial lymphatic network in the tails of 20 nude mice. As described previously (17) , 5 µl of 25% FITC-dextran solution were injected at the tail tip, and progressive staining of the superficial capillary network of the tail, visible by low-power fluorescence microscopy, was detected by an intensified charge-coupled device video camera (model 2400; Hamamatsu Photonics, Hamamatsu, Japan) and recorded on videotape. On day 2, a suspension of mouse fibrosarcoma cells (FSaII) was injected intradermally 1–2 cm from the tip of the tail in 15 mice, whereas the remaining control group of 5 mice received a sham injection of saline. On day 30, the fluorescence microlymphography procedure was performed again on all mice. On day 31, the lymphatic staining procedure was repeated using 5 µl of ferritin solution (108 mg/ml in 0.15 M NaCl; filter sterilized) in place of the fluorescent dextran. In five of the tumor-bearing mice, the ferritin solution was injected directly into the center of the tumor rather than at the tail tip. Forty-five min after injection, the mouse was sacrificed, and the tail was amputated and prepared for histology. The distal portion of each tail was fixed in 4% formalin and cut into 12 pieces of 0.5 cm length. Thin 3-µm sections were cut from each large section embedded in historesin. The sections were stained for light microscopy with H&E for tissue contrast and with Prussian Blue (potassium ferrocyanide and HCl) to reveal the iron III component of ferritin.

In Situ Hybridization.
In situ hybridization studies were performed as described previously (9 , 11 , 21) . Thin sections were cut from the paraffin-embedded tail and then hybridized with an antisense probe for VEGF-C, VEGFR-2, or VEGFR-3. Hybridization with the sense probe for VEGFR-3 was performed as a control to establish the background signal.


   Results and Discussion
Top
 ABSTRACT
 Introduction
 Materials and Methods
 Results and Discussion
REFERENCES
 
The microlymphography technique relies on the principle that locally injected high molecular weight agents (ferritin and FITC-dextran) are eliminated from the tissue interstitium via the lymphatics. The tails of 20 nude mice were examined by fluorescence microlymphography to detect the functional superficial lymphatic network in the skin before and 28 days after intradermal implantation of FSaII murine sarcoma cells (15 mice) or saline (5 mice).

Fluorescence microlymphography on the day before tumor injection showed progressive staining of a mesh-like capillary network extending the full length of the tail in all animals (Fig. 1ACitation ). Twenty-eight days after implantation, tumors had developed in 11 tails, with a mean tumor diameter ± SD of 1.14 ± 1.0 cm and a mean distance from the tail tip of 0.98 ± 0.90 cm. Fluorescence microlymphography confirmed that the network remained intact in the control group. Tumor-bearing animals showed only a partial lymphatic network with a notable absence of staining in skin overlying the tumor area (Fig. 1BCitation ). By injecting FITC-dextran at high pressure into the dermis near the tumor, these lymphatics will fill. This implicates the local mechanics of the tumor in the alteration of lymphatic function (22) . Dilated lymphatic vessels were observed near the tumor border, but the position of these vessels relative to the true tumor boundary could only be determined by subsequent histological examination.



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  		Fig. 1. Fluorescence microlymphography of the nude mouse tail after the injection of 5 µl of FITC-dextran (Mr 2,000,000; 25% solution) at the distal tip. The direction of lymph flow is from left to right. Bar, 400 µm. A, typical continuous network, 22 min after injection. B, the same tail, 28 days after injection of a FSaII cell suspension (1.5 x 106 cells in 10 µl of phosphate-buffered saline). The network did not stain in the skin above the tumor area (right side of micrograph); fluorescent staining shows that the surrounding lymph capillaries remain functional. Vessels at the tumor border appear dilated and possibly leaky and discontinuous. Large arrows indicate attenuated vessels possibly inside the tumor. Small arrows indicate the increased apparent diameter due to engorgement and/or flattening of the lymphatic capillary.

 
Histological/functional examination was performed 29 days after tumor implantation using ferritin as a lymph marker. Mice were sacrificed 45 min after intradermal injection of ferritin at the tail tip or directly into the tumor. The tails were processed for histology and stained with Prussian Blue for detection of ferritin by light microscopy. Cross-sections through normal tissue reveal the lymphatic vessels at various depths in the extrafascial layer, always above the fascia muscularis (Fig. 2ACitation ). Occasionally, structures that appear to be unstained lymphatic vessels were observed, judging by the thin irregular walls enclosing amorphous fluid, lipid droplets, and lymphocytes, but no erythrocytes.



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  		Fig. 2. Cross-sections through mouse tail prepared 45 min after in vivo administration of ferritin. A, in tumor-free tissue, several prefascial lymphatics are visible (marked by arrows), corresponding to the superficial network seen by fluorescence microlymphography. These are stained dark blue due to the presence of ferritin in the lymphatic fluid. Typical lipid droplets are also visible. Normal lymphatics generally had a flattened appearance in the cross-section. Lighter blue stain outside the lymphatics is most likely due to endogenous ferritin in connective tissue. At this magnification (x100), the bar denotes 200 µm. B, in a tumor-bearing tail, several prefascial lymphatics (arrow) situated in the region between the tumor (below and to the right of arrow) and skin (above the arrow) appear dilated. Magnification, x200. C, lymphatic vessel at the periphery. Tumor tissue is directly below the vessel and partially surrounds it. Several tumor cells can be identified within the lumen (arrows). Magnification, x400. D, two lymphatic vessels with partly intact endothelial cells are completely surrounded by tumor cells. The boundary with normal tissue is at the top right half of the micrograph. The top vessel contains a tumor cell (arrow). Magnification, x400.

 
Solid tumor tissue surrounded the bone (either completely or asymmetrically) and remained outwardly confined by the intact epidermis without invasion into those structures (Fig. 2BCitation ). Within the tumor, normal tissue structures were not evident, with the exception of occasional ferritin-stained lymphatics observed between the tumor and epidermal layer and within peripheral areas of the tumor. No ferritin-stained vessels were encountered within the tumor interior (defined as greater than 100 µm inside the tumor boundary) in any animal, even those that received a direct intratumoral ferritin injection. However, 6 of 11 tumors had at least one functional lymphatic vessel at the periphery that was partially (Fig. 2CCitation ) or completely surrounded by tumor cells (Fig. 2DCitation ). Other vessels appeared to be intact, with a distinct endothelial lining (Fig. 2DCitation ). Single tumor cells and clumps were present within the lumina of these intratumoral lymphatics.

Table 1Citation summarizes the observations made on the control group of five mice and the two groups of tumor-bearing mice that received ferritin injections either at the tail tip (six mice) or directly into the tumor (five mice). Direct intratumoral injection of ferritin did not reveal the existence of deeper lymphatic vessels. Ferritin-labeled lymph appeared in normal tissue lymphatics distal to the tumor and, in most cases, proximal to the tumor, but a lymphatic vessel was detected in only one of the five directly injected tumors, and that one was at the periphery.


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  		Table 1 Summary of observations in 16 mice

The results of fluorescence microlymphography and histology are tabulated. Vessel dilation and staining were judged to be either strong (++), weak (+), or negative. Other observations were either positive or negative. The tumor periphery was defined as the portion of the tumor less than 100 µm distant from the interface with normal tissue, and the interior portion of the tumor was defined as the portion more than 100 µm from normal tissue.

 
A separate group of tumors was processed for detection of VEGFR-2 (Flk-1), VEGFR-3 (FLT4), and VEGF-C by in situ hybridization. Similar to the findings in ferritin-stained sections, increased numbers of vessels within the tumor periphery were enlarged. The expression of both receptors (VEGFR-2 and VEGFR-3) in tumors was more intense than that seen in the normal dermis. Most areas of intense VEGFR-3 expression corresponded with lymphatic vessels in the dermis surrounding the tumor (Fig. 3 and BCitation ) and within the tumor periphery (Fig. 3 and DCitation ). However, there were other areas of strong VEGFR-3 expression in the tumor interior that could not be positively identified as lymphatic structures, although many of the VEGFR-3-expressing cells were endothelial cells (8) . In contrast to VEGFR-3-hybridization, VEGFR-2 hybridization (Fig. 3 and FCitation ) revealed high expression in vascular structures throughout the tumor. This was consistent with the known role of VEGFR-2 as a mitogenic receptor involved in hemangiogenesis. Expression of VEGF-C (Fig. 3 and HCitation ) was more intense than the background level (Fig. 3ICitation ) and appeared localized to clusters of cells. Because lymphatic vessels were not discernible within the interior of the tumor, it was not clear whether these clusters corresponded to lymphatic endothelial cells.



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  		Fig. 3. In situ hybridization analysis of VEGFR-3, VEGFR-2, and VEGF-C expression in FSaII tumor and the surrounding dermis in the tail. A, C, E, and G show antisense mRNA probe for VEGFR-3 (A and C), VEGFR-2 (E), and VEGF-C (G), whereas I shows the background level of signal obtained using the VEGFR-3 sense probe. B, D, F, H, and J are the corresponding counterstained sections of tail. A and B, intense VEGFR-3 expression in lymphatics of the normal dermis at the periphery of the tumor. Arrows, lymphatic vessels. C and D, the VEGFR-3 probe shows strong expression in vessels at the periphery of the tumor. In the tumor interior, clusters of strong VEGFR-3 expression do not appear to be vessels, although at least some of the cells are endothelial. E and F, all vessels within the tumor (mostly blood vessels in the tumor interior) show strong expression of VEGFR-2 compared with the surrounding tissue. G and H, staining by VEGF-C probe is more intense than control staining (VEGFR-3 sense probe, I and J) and appears to be localized in groups of cells, although not with any identifiable vessels. Bar in I, 100 µm.

 
The results of this combined in vivo and histological study support a model in which the solid tumor is largely devoid of functional lymphatic capillaries but is surrounded by functional lymphatics in the adjacent normal tissue and evidently in the tumor periphery. The combined methods failed to detect any functional lymphatics within the bulk of the tumor. This lack of intratumoral functional lymphatics may explain the enlarged state of lymphatic vessels surrounding the tumor because the tumor interstitial fluid formed by the net filtration of plasma from the tumor vasculature must ultimately seep out of the tumor into the adjacent tissue (1 , 23) . The dilated appearance of some vessels at the tumor/normal interface suggests increased drainage activity of the functional lymphatics there, although this could not be quantified. Given the abundant expression of VEGFR-3, it is also possible that the increased lymphatic vessel diameter in this region reflects endothelial cell proliferation induced by VEGF-C (11) . Additionally, the high-pressure intradermal lymphangiography shown in Padera et al. (22) illustrates that peritumor lymphatics maintain the regular network pattern seen in normal tail skin, suggesting that these lymphatics are not new lymphatics but existing ones. Based on these observations, it appears that the engorged peritumor lymphatics shown here are likely to be existing vessels and not new lymphatics. Thus, it is unnecessary to invoke the phenomenon of tumor lymphangiogenesis to account for lymphatic metastasis; the observed engorgement of lymph vessels at the tumor periphery would seem to offer ample opportunity for dissemination of potentially metastatic cells.

Given the presence of VEGFR-3, we tested for the presence of its ligand, VEGF-C, and detected its expression at a modest level within the tumor. It is unclear whether this constitutes a lymphangiogenic signal or whether higher levels of VEGF-C would be required to stimulate the development of lymphatics in the tumor. Based on the evidence to date, we propose that destruction of lymphatics in the tumor is due to collapse under the pressure (mechanical stress) of growing cancer cells. Proliferating tumor cells can generate a solid stress of 45–120 mm Hg when grown as spheroids in vitro (24) , a solid stress sufficient to collapse a tube of endothelial cells. This leads to an interesting paradox: why are many tumor blood vessels open and functional, whereas most lymphatic vessels are collapsed? Our hypothesis is that the connection of blood vessels to the high-pressure arterial blood supply prevents collapse, whereas lymphatic vessels have no comparable high-pressure source. Whereas this mechanical hypothesis seems reasonable, the presence of an antilymphangiogenic molecule(s) in tumors cannot be excluded.


   Acknowledgments
 
We are indebted to Drs. H. J. Leu, B. Odermatt, and J. Schneider of the Institute of Pathology, University Hospital Zurich (Zurich, Switzerland) for histopathological advice. We thank Drs. Y. Boucher, S. Patan, M. Swartz, M. Endo, H. Lichtenbeld, and C. Mouta Carreira for helpful discussions and T. P. Padera for invaluable input and help with preparation of the manuscript.


   FOOTNOTES
 
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1 Supported by Outstanding Investigator Grant R35-CA-56591 from the National Cancer Institute (to R. K. J.), a grant from the National Foundation for Cancer Research (to R. K. J.), a fellowship from the Swiss National Science Foundation (to A. J. L.), and a Biomedical Engineering Research grant from the Whitaker Foundation (to D. A. B.). A. J. L. and D. A. B. contributed equally to this work. Back

2 Present address: Gefäss-Zentrum, Klinik Hirslanden, Witellikerstrasse 40, 8008 Zürich, Switzerland. Back

3 Present address: School of Pharmacy and Pharmaceutical Sciences, University of Manchester, Oxford Road, Manchester M13 9PL, United Kingdom. Back

4 To whom requests for reprints should be addressed, at Department of Radiation Oncology, Massachusetts General Hospital, 100 Blossom Street, Cox-7, Boston, MA 02114. E-mail: jain{at}steele.mgh.harvard.edu Back

5 The abbreviations used are: VEGFR, vascular endothelial growth factor receptor; VEGF, vascular endothelial growth factor. Back

Received 5/ 8/00. Accepted 6/29/00.


   REFERENCES
Top
 ABSTRACT
 Introduction
 Materials and Methods
 Results and Discussion
 REFERENCES
 

  1. Gullino P. M. Angiogenesis, tumor vascularization, and potential interference with tumor growth Mihich E. eds. . Biological Responses in Cancer, 4: 1-20, Plenum Press New York 1985.
  2. Folkman J. Angiogenesis and tumor growth. N. Engl. J. Med., 334: 921-926, 1996.
  3. Lee F. C., Tilghmanm R. C. Lymph vessels in rabbit carcinoma, with a note on the normal lymph vessel structure of the testis. Arch. Surg., 26: 602-616, 1933.[Abstract/Free Full Text]
  4. Pullinger B. D., Florey H. W. Proliferation of lymphatics in inflammation. J. Pathol. Biol., 45: 157-170, 1937.
  5. Zeidman I., Copeland B. E., Warren S. Experimental studies on the spread of cancer in the lymphatic system. II. Absence of a lymphatic supply in carcinomas. Cancer (Phila.), 8: 123-127, 1955.
  6. Gilchrist R. K. Surgical management of advanced cancer of the breast. Arch. Surg., 61: 913-929, 1950.
  7. Tanigawa N., Kanazawa T., Satomura K., Hikasa Y., Hashida M., Muranishi S., Sezaki H. Experimental study on lymphatic vascular changes in the development of cancer. Lymphology, 14: 149-154, 1981.[Medline]
  8. Partanen T. A., Alitalo K., Miettinen M. Lack of lymphatic vascular specificity of vascular endothelial growth factor receptor 3 in 185 vascular tumors. Cancer (Phila.), 86: 2406-2412, 1999.[Medline]
  9. Joukov V., Pajusola K., Kaipainen A., Chilov D., Lahtinen I., Kukk E., Saksela O., Kalkkinen N., Alitalo K. A novel vascular endothelial growth factor, VEGF-C, is a ligand for the Flt4 (VEGFR-3) and KDR (VEGFR-2) receptor tyrosine kinases. EMBO J., 15: 290-298, 1996.[Medline]
  10. Lee J., Gray A., Yuan J., Luoh S. M., Avraham H., Wood W. I. Vascular endothelial growth factor-related protein: a ligand and specific activator of the tyrosine kinase receptor Flt4. Proc. Natl. Acad. Sci. USA, 93: 1988-1992, 1996.[Abstract/Free Full Text]
  11. Jeltsch M., Kaipainen A., Joukov V., Meng X., Lakso M., Rauvala H., Swartz M., Fukumura D., Jain R. K., Alitalo K. Hyperplasia of lymphatic vessels in VEGF-C transgenic mice. Science (Washington DC), 276: 1423-1425, 1997.[Abstract/Free Full Text]
  12. Jussila L., Valtola R., Partanen T. A., Salven P., Heikkila P., Matikainen M. T., Renkonen R., Kaipainen A., Detmar M., Tschachler E., Alitalo R., Alitalo K. Lymphatic endothelium and Kaposi’s sarcoma spindle cells detected by antibodies against the vascular endothelial growth factor receptor-3. Cancer Res., 58: 1599-1604, 1998.[Abstract/Free Full Text]
  13. Lymboussaki A., Partanen T. A., Olofsson B., Thomas-Crusells J., Fletcher C. D., de Waal R. M., Kaipainen A., Alitalo K. Expression of the vascular endothelial growth factor C receptor VEGFR-3 in lymphatic endothelium of the skin and in vascular tumors. Am. J. Pathol., 153: 395-403, 1998.[Abstract/Free Full Text]
  14. Salven P., Lymboussaki A., Heikkila P., Jaaskela-Saari H., Enholm B., Aase K., von Euler G., Eriksson U., Alitalo K., Joensuu H. Vascular endothelial growth factors VEGF-B and VEGF-C are expressed in human tumors. Am. J. Pathol., 153: 103-108, 1998.[Abstract/Free Full Text]
  15. Valtola R., Salven P., Heikkila P., Taipale J., Joensuu H., Rehn M., Pihlajaniemi T., Weich H., deWaal R., Alitalo K. VEGFR-3 and its ligand VEGF-C are associated with angiogenesis in breast cancer. Am. J. Pathol., 154: 1381-1390, 1999.[Abstract/Free Full Text]
  16. Yonemura Y., Endo Y., Fujita H., Fushida S., Ninomiya I., Bandou E., Taniguchi K., Miwa K., Ohoyama S., Sugiyama K., Sasaki T. Role of vascular endothelial growth factor C expression in the development of lymph node metastasis in gastric cancer. Clin. Cancer Res., 5: 1823-1829, 1999.[Abstract/Free Full Text]
  17. Leu A. J., Berk D. A., Yuan F., Jain R. K. Flow velocity in the superficial lymphatic network of the mouse tail. Am. J. Physiol., 267: H1507-H1513, 1994.[Abstract/Free Full Text]
  18. Swartz M. A., Berk D. A., Jain R. K. Transport in lymphatic capillaries. I. Macroscopic measurements using residence time distribution theory. Am. J. Physiol., 270: H324-H329, 1996.
  19. Berk D. A., Swartz M. A., Leu A. J., Jain R. K. Transport in lymphatic capillaries. II. Microscopic velocity measurement with fluorescence photobleaching. Am. J. Physiol., 270: H330-H337, 1996.
  20. Bollinger A., Jager K., Sgier F., Seglias J. Fluorescence microlymphography. Circulation, 64: 1195-200, 1981.[Abstract/Free Full Text]
  21. Kaipainen A., Korhonen J., Mustonen T., van Hinsbergh V. W., Fang G. H., 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. USA, 92: 3566-3570, 1995.[Abstract/Free Full Text]
  22. Padera T. P., Yun C., Kadambi A., Mouta Carreira, C., Jain R. K. Local mechanics and VEGF-C alter peri-tumor lymphatic function. Proc. Am. Assoc. Cancer Res., 41: 88 2000.
  23. Jain R. K. Delivery of novel therapeutic agents in tumors: physiological barriers and strategies. J. Natl. Cancer Inst., 81: 570-576, 1989.[Free Full Text]
  24. Helmlinger G., Netti P. A., Lichtenbeld H. C., Melder R. J., Jain R. K. Solid stress inhibits the growth of multicellular tumor spheroids. Nat. Biotechnol., 15: 778-783, 1997.[Medline]



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Home page
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P. Laakkonen, M. E. Akerman, H. Biliran, M. Yang, F. Ferrer, T. Karpanen, R. M. Hoffman, and E. Ruoslahti
Antitumor activity of a homing peptide that targets tumor lymphatics and tumor cells
PNAS, June 22, 2004; 101(25): 9381 - 9386.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
R. T. Tong, Y. Boucher, S. V. Kozin, F. Winkler, D. J. Hicklin, and R. K. Jain
Vascular Normalization by Vascular Endothelial Growth Factor Receptor 2 Blockade Induces a Pressure Gradient Across the Vasculature and Improves Drug Penetration in Tumors
Cancer Res., June 1, 2004; 64(11): 3731 - 3736.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
T. P. Padera, Y. Boucher, R. K. Jain, L. M. Wein, P. E. Holden, and D. H. Kirn
Correspondence re: S. Maula et al., Intratumoral Lymphatics Are Essential for the Metastatic Spread and Prognosis in Squamous Cell Carcinoma of the Head and Neck. Cancer Res., 63: 1920-1926, 2003.
Cancer Res., December 1, 2003; 63(23): 8555 - 8557.
[Full Text] [PDF]

Home page
 JCBHome page
M. S. Pepper and M. Skobe
Lymphatic endothelium: morphological, molecular and functional properties
J. Cell Biol., October 27, 2003; 163(2): 209 - 213.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Pathol.Home page
S. S. Dadras, T. Paul, J. Bertoncini, L. F. Brown, A. Muzikansky, D. G. Jackson, U. Ellwanger, C. Garbe, M. C. Mihm, and M. Detmar
Tumor Lymphangiogenesis: A Novel Prognostic Indicator for Cutaneous Melanoma Metastasis and Survival
Am. J. Pathol., June 1, 2003; 162(6): 1951 - 1960.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
J. Krishnan, V. Kirkin, A. Steffen, M. Hegen, D. Weih, S. Tomarev, J. Wilting, and J. P. Sleeman
Differential in Vivo and in Vitro Expression of Vascular Endothelial Growth Factor (VEGF)-C and VEGF-D in Tumors and Its Relationship to Lymphatic Metastasis in Immunocompetent Rats
Cancer Res., February 1, 2003; 63(3): 713 - 722.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
H. Kobayashi, S. Kawamoto, R. A. Star, T. A. Waldmann, Y. Tagaya, and M. W. Brechbiel
Micro-magnetic Resonance Lymphangiography in Mice Using a Novel Dendrimer-based Magnetic Resonance Imaging Contrast Agent
Cancer Res., January 15, 2003; 63(2): 271 - 276.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
H. Wiig, K. Aukland, and O. Tenstad
Isolation of interstitial fluid from rat mammary tumors by a centrifugation method
Am J Physiol Heart Circ Physiol, January 1, 2003; 284(1): H416 - H424.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
H. Dafni, T. Israely, Z. M. Bhujwalla, L. E. Benjamin, and M. Neeman
Overexpression of Vascular Endothelial Growth Factor 165 Drives Peritumor Interstitial Convection and Induces Lymphatic Drain: Magnetic Resonance Imaging, Confocal Microscopy, and Histological Tracking of Triple-labeled Albumin
Cancer Res., November 15, 2002; 62(22): 6731 - 6739.
[Abstract] [Full Text] [PDF]

Home page
 JEMHome page
M. Detmar and S. Hirakawa
The Formation of Lymphatic Vessels and Its Importance in the Setting of Malignancy
J. Exp. Med., September 16, 2002; 196(6): 713 - 718.
[Full Text] [PDF]

Home page
 Am. J. Pathol.Home page
S. F. Schoppmann, P. Birner, J. Stockl, R. Kalt, R. Ullrich, C. Caucig, E. Kriehuber, K. Nagy, K. Alitalo, and D. Kerjaschki
Tumor-Associated Macrophages Express Lymphatic Endothelial Growth Factors and Are Related to Peritumoral Lymphangiogenesis
Am. J. Pathol., September 1, 2002; 161(3): 947 - 956.
[Abstract] [Full Text] [PDF]

Home page
 FASEB J.Home page
S. A. STACKER, M. E. BALDWIN, and M. G. ACHEN
The role of tumor lymphangiogenesis in metastatic spread
FASEB J, July 1, 2002; 16(9): 922 - 934.
[Abstract] [Full Text] [PDF]

Home page
 Physiol. Rev.Home page
L. Jussila and K. Alitalo
Vascular Growth Factors and Lymphangiogenesis
Physiol Rev, July 1, 2002; 82(3): 673 - 700.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
H. Kubo, R. Cao, E. Brakenhielm, T. Makinen, Y. Cao, and K. Alitalo
Blockade of vascular endothelial growth factor receptor-3 signaling inhibits fibroblast growth factor-2-induced lymphangiogenesis in mouse cornea
PNAS, June 25, 2002; 99(13): 8868 - 8873.
[Abstract] [Full Text] [PDF]

Home page
 ScienceHome page
T. P. Padera, A. Kadambi, E. di Tomaso, C. M. Carreira, E. B. Brown, Y. Boucher, N. C. Choi, D. Mathisen, J. Wain, E. J. Mark, et al.
Lymphatic Metastasis in the Absence of Functional Intratumor Lymphatics
Science, June 7, 2002; 296(5574): 1883 - 1886.
[Abstract] [Full Text] [PDF]

Home page
 JNCI J Natl Cancer InstHome page
R. K. Jain and T. P. Padera
Prevention and Treatment of Lymphatic Metastasis by Antilymphangiogenic Therapy
J Natl Cancer Inst, June 5, 2002; 94(11): 785 - 787.
[Full Text] [PDF]

Home page
 JNCI J Natl Cancer InstHome page
Y. He, K.-i. Kozaki, T. Karpanen, K. Koshikawa, S. Yla-Herttuala, T. Takahashi, and K. Alitalo
Suppression of Tumor Lymphangiogenesis and Lymph Node Metastasis by Blocking Vascular Endothelial Growth Factor Receptor 3 Signaling
J Natl Cancer Inst, June 5, 2002; 94(11): 819 - 825.
[Abstract] [Full Text] [PDF]

Home page
 JNCI J Natl Cancer InstHome page
R. K. Jain and B. T. Fenton
Intratumoral Lymphatic Vessels: A Case of Mistaken Identity or Malfunction?
J Natl Cancer Inst, March 20, 2002; 94(6): 417 - 421.
[Full Text] [PDF]

Home page
 Cancer Res.Home page
N. J. P. Beasley, R. Prevo, S. Banerji, R. D. Leek, J. Moore, P. van Trappen, G. Cox, A. L. Harris, and D. G. Jackson
Intratumoral Lymphangiogenesis and Lymph Node Metastasis in Head and Neck Cancer
Cancer Res., March 1, 2002; 62(5): 1315 - 1320.
[Abstract] [Full Text] [PDF]

Home page
 Cold Spring Harb Symp Quant BiolHome page
R.K. JAIN
Angiogenesis and Lymphangiogenesis in Tumors: Insights from Intravital Microscopy
Cold Spring Harb Symp Quant Biol, January 1, 2002; 67(0): 239 - 248.
[Abstract] [PDF]

Home page
 Cancer Res.Home page
C. M. Carreira, S. M. Nasser, E. di Tomaso, T. P. Padera, Y. Boucher, S. I. Tomarev, and R. K. Jain
LYVE-1 Is Not Restricted to the Lymph Vessels: Expression in Normal Liver Blood Sinusoids and Down-Regulation in Human Liver Cancer and Cirrhosis
Cancer Res., November 1, 2001; 61(22): 8079 - 8084.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
M. Milosevic, A. Fyles, D. Hedley, M. Pintilie, W. Levin, L. Manchul, and R. Hill
Interstitial Fluid Pressure Predicts Survival in Patients with Cervix Cancer Independent of Clinical Prognostic Factors and Tumor Oxygen Measurements
Cancer Res., September 1, 2001; 61(17): 6400 - 6405.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Pathol.Home page
M. Skobe, L. M. Hamberg, T. Hawighorst, M. Schirner, G. L. Wolf, K. Alitalo, and M. Detmar
Concurrent Induction of Lymphangiogenesis, Angiogenesis, and Macrophage Recruitment by Vascular Endothelial Growth Factor-C in Melanoma
Am. J. Pathol., September 1, 2001; 159(3): 893 - 903.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
T. Karpanen, M. Egeblad, M. J. Karkkainen, H. Kubo, S. Ylä-Herttuala, M. Jäättelä, and K. Alitalo
Vascular Endothelial Growth Factor C Promotes Tumor Lymphangiogenesis and Intralymphatic Tumor Growth
Cancer Res., March 1, 2001; 61(5): 1786 - 1790.
[Abstract] [Full Text]

Home page
 Cancer Res.Home page
A. Kadambi, C. Mouta Carreira, C.-o. Yun, T. P. Padera, D. E. J. G. J. Dolmans, P. Carmeliet, D. Fukumura, and R. K. Jain
Vascular Endothelial Growth Factor (VEGF)-C Differentially Affects Tumor Vascular Function and Leukocyte Recruitment: Role of VEGF-Receptor 2 and Host VEGF-A
Cancer Res., March 1, 2001; 61(6): 2404 - 2408.
[Abstract] [Full Text]

Home page
 Clin. Cancer Res.Home page
M. S. Pepper
Lymphangiogenesis and Tumor Metastasis: Myth or Reality?
Clin. Cancer Res., March 1, 2001; 7(3): 462 - 468.
[Abstract] [Full Text]

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