Elsevier

Lung Cancer

Volume 44, Issue 2, May 2004, Pages 167-174
Lung Cancer

Influence of basic fibroblast growth factor on the proliferation of non-small cell lung cancer cell lines

https://doi.org/10.1016/j.lungcan.2003.11.005Get rights and content

Abstract

Basic fibroblast growth factor (bFGF) is closely involved in angiogenesis and tumor growth of various cancers, but its role in proliferation and differentiation of non-small cell lung cancer (NSCLC) remains to be defined. The majority of NSCLC cell lines produce elevated protein levels of bFGF but do not secrete comparable amounts. We therefore investigated the influence of bFGF on the proliferation of three human NSCLC cell lines. Our experiments demonstrate that intracellular bFGF level and bFGF mRNA expression correlated with the proliferation rate in all three cell lines. Delivery of a bFGF neutralizing monoclonal antibody, anti-sense oligonucleotides or a vector expressing bFGF antisense cDNA into the cells inhibited tumor cell growth. Delivery of recombinant bFGF into a bFGF-negative cell line led to increased proliferation. These findings suggest that bFGF stimulates the growth of tumor cells by intracrine mechanisms. Strategies to inhibit bFGF in NSCLC may therefore be a promising approach in NSCLC therapy.

Introduction

Non-small cell lung cancer (NSCLC) is one of the leading cause of cancer-related death in the western world. Despite the development of new chemotherapeutic agents, and progress in surgical and radiotherapy techniques, survival rates of lung cancer patients have hardly changed. A better understanding of biological events leading to tumor progression will eventually contribute to the development of novel therapeutic strategies.

Tumor growth and progression are based on a complex interaction between tumor and tumor-associated tissue which is mediated, e.g. by several growth factors. Basic fibroblast growth factor (bFGF), a member of a group of heparin-binding multifunctional polypeptides, is one of the most potent angiogenic factors [1], [2]. It is also involved in proliferation and differentiation of a variety of normal and malignant cells and tissues [3], [4]. However, the precise role of bFGF in NSCLC development and progression is unclear. Several authors have reported a correlation of bFGF expression as well as bFGF serum levels with progression and prognosis of NSCLC [5], [6], [7], [8]. Other investigators have not found any correlation between bFGF immunoreactivity of cancer cells or serum levels of bFGF and prognosis [9], [10]. In fact, Guddo and colleagues have reported that bFGF expression is inversely correlated with tumor progression [11].

Nevertheless, elevated levels of bFGF were produced in almost all NSCLC cells in vitro and in vivo, and the in vitro bFGF production correlated with the doubling time of NSCLC cell lines [12]. To clarify the role of bFGF in the growth of NSCLC, we investigated bFGF expression in NSCLC cell lines, as well as proliferation of these cell lines following the modulation of intracellular bFGF level.

Section snippets

Cell lines and cell culture

All cell lines used were purchased from ATCC. The lung cancer (NSCLC) cell lines H1299 (ATCC-Nr.: CRL-5803; morphology: epithelial) H460 (HTB-177; epithelial), A549 (CCL-185; epithelial) and H358 (CRL-5807; epithelial) were maintained in RPMI 1640, supplemented with 10% FCS v/v (fetal calf serum) and 2 mM glutamine at 37 °C, 5% CO2 v/v in a humidified atmosphere. For experiments, cells were seeded, treated and analyzed as indicated.

Determination of the proliferation rate

Cells were counted at 24, 48, 72, and 96 h after each treatment.

Intracellular bFGF expression and proliferation of NSCLC cell lines

The proliferation rates of cell lines H460, H1299 and A549 were determined at 24, 48, 72, and 96 h. At each time point the bFGF levels were determined in cell extracts and in culture supernatants. The proliferation rate was found to be increased during the first 48 h in all cell lines (Fig. 1A). After cells reached confluency proliferation rate fell in all cell lines except H1299 cells. These cells exhibited a decline in proliferation rate only at times later than 72 h. A similar time course was

Discussion

The function of bFGF as a potent angiogenesis stimulating molecule has been described in detail [6], [13]. The role of intracellular bFGF in tumor growth however has not been elucidated. In human colorectal carcinoma cells bFGF promotes the process of tumor cell invasion [16]. In several tumours such as melanoma or bladder cancer the suppression of bFGF led to inhibition of tumor growth [14], [15]. On the other hand Fenig and colleagues have reported that bFGF selectively inhibits proliferation

Acknowledgements

This work was supported in part by the Saxonian Department of Environment and Agriculture (SMUL, Germany) Az. 13-8811.61/102. The authors would like acknowledge the expert technical assistance of Constanze Büttner.

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