Cancer Letters

Cancer Letters

Volume 249, Issue 2, 8 May 2007, Pages 188-197
Cancer Letters

Hypoxia influences vasculogenic mimicry channel formation and tumor invasion-related protein expression in melanoma

https://doi.org/10.1016/j.canlet.2006.08.016Get rights and content

Abstract

Background

Hypoxia can enhance tumor cell invasion and metastasis. The cause and the molecular mechanism are still not clear.

Methods

In our study, mouse melanoma B16 cells were inoculated into mouse ischemic limbs and non-ischemic controls and the engrafted melanomas were subsequently observed. Vasculogenic mimicry channels in melanoma tumors of the two groups were counted and the expression of HIF-1α, MMP-2, MMP-9 and VEGF was assessed by immunohistochemical staining. Formalin-fixed, paraffin-embedded tissues were used for immunohistochemical staining.

Results

In the early stage of engrafted melanoma growth, the size of melanomas in ischemic limbs increased slower than in the controls. However, later there was no obvious difference in their size. Melanoma tumors in the ischemic group had more vasculogenic mimicry channels than those in the controls (P = 0.039). Similarly, the expression of HIF-1α, MMP-2, MMP-9 and VEGF was higher in the ischemic group than in the non-ischemic controls (P = 0.024, 0.047, 0.007 and 0.025, respectively). There was a positive association in melanoma cells of the ischemic group between expression of HIF-1α and VEGF, and also between MMP-9 and MMP-2. In the ischemic group, there was statistical significance for the correlation between HIF-1α and VEGF expression (r = 0.456, P = 0.038). Furthermore, MMP-2 expression was positively correlated with MMP-9 and VEGF expression (r = 0.589 and 0.502, P = 0.008 and 0.024, respectively).

Conclusions

Melanoma cells in a hypoxic microenvironment increased HIF-1α expression and induced the formation of vasculogenic mimicry channels to acquire an adequate blood supply. On the other hand, the expression of MMP-2 and MMP-9 in tumor tissue increased to enhance the invasiveness. HIF-1α, MMP-2 and MMP-9 may be associated with the failure of stop-flow perfusion in some patients with melanoma.

Section snippets

Background

It is well known that melanoma is a type of aggressive malignant tumor with poor prognosis [1]. Pilati and Guadagni et al. [2], [3] reported a novel therapy, namely hypoxic antiblastic stop-flow perfusion (SFP), for patients with limb-sited melanoma. In this method, chemical agents are perfused into the melanoma through a large artery on the tumor-bearing limb under hypoxic conditions. However, it is not as effective as expected for some patients. Hypoxia is a two-edged sword for tumor growth.

Cells

Mouse melanoma B16 cells were supplied by Tianjin Cancer Hospital. Melanoma single cell suspensions, which had been stored in liquid nitrogen, were incubated in a 40 °C water bath for 10 min prior to injection. The cells were counted and diluted to 1 × 106/ml with normal solution buffer for the injections.

Animal model

All procedures were performed on 7-week-old C57 mice, including 20 males and 20 females. The forty mice were randomly divided into two groups, group A and group B, with 20 mice per group. Ischemia

The ischemic hindlimb and the growth of the engrafted melanoma

Twelve days after surgery, the temperature of the ischemic limbs was lower than the normal limbs and the ischemic limbs were unable to move. By this time, the surgical wounds had healed without visible infection. The engrafted melanomas could be palpated on the non-ischemic limb on the 7th day following tumor cell injection, while the tumor in the ischemic limbs could not be confirmed by palpation until the 9th day. When the mice were sacrificed on the 15th day, a black tumor mass was observed

Discussion

Melanoma is a type of malignant and poorly differentiated tumor that is prone to metastasize and the prognosis of the patients is very poor [1], [14], [15]. There are numerous treatments for melanoma depending on tumor location. Hypoxic antiblastic SFP, aimed at limb-sited melanoma, was recently reported by Rossi CR. and his colleagues [2], [3]. In this therapy, chemical agents such as TNF-α and melphalan are perfused into the local melanoma tissue via the local artery when the blood flow is

Acknowledgements

This work was partly supported by a grant from the National Nature Science Foundation of China (No. 30370554). We are grateful to Valerie Dunmire for assistance with English amendment.

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    These authors contributed equally to this work.

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