VCAM-1 secreted from cancer-associated fibroblasts enhances the growth and invasion of lung cancer cells through AKT and MAPK signaling

doi:10.1016/j.canlet.2019.12.039

Cancer Letters

Volume 473, 31 March 2020, Pages 62-73

https://doi.org/10.1016/j.canlet.2019.12.039 Get rights and content

Highlights

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VCAM-1secreted from cancer associated fibroblasts (CAFs) promotes lung cancer cells proliferation, migration and invasion.
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CAFs activated AKT and MAPK signaling via VCAM-1 receptor very-late antigen (VLA)-4 in lung cancer cells.
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VCAM-1 expression was associated with a poor prognosis in lung cancer patients.

Abstract

Several studies have indicated that cancer-associated fibroblasts (CAFs) could promote cancer progression in many malignancies. However, the mechanism by which CAFs promote the growth and metastasis of lung cancer remains poorly defined. In the present study, CAFs and normal fibroblasts (NFs) were isolated from human lung cancer and adjacent tissue. The data showed that the conditional medium (CM) of CAFs could increase the proliferation, migration and invasion of lung cancer cells. Vascular cell adhesion molecule-1 (VCAM-1) showed a higher expression in CAF-CM than NF-CM, and blocking VCAM-1 in CAF-CM attenuated the proliferation and invasion of cancer cells. Further, the results showed that VCAM-1 secreted from CAFs activated AKT and MAPK signaling via receptor α4β1 integrin (very-late antigen (VLA)-4) in lung cancer cells. Moreover, CAFs promoted VCAM-1 expression and tumor growth in vivo. Additionally, bioinformatics analysis indicated a positive correlation on the CAF marker protein alpha-smooth muscle actin (α-SMA) and VCAM-1 expression, which was associated with a poor prognosis in lung cancer patients. These findings demonstrate that the VCAM-1 secreted from CAFs enhances growth and invasion by activating the AKT and MAPK signaling of lung cancer cells.

Introduction

Lung cancer is the leading cause of cancer death and exhibits the highest morbidity and mortality globally, the overall 5-year survival rate is 15% [1,2]. Most lung cancer patients die of metastasis, thus, understanding the mechanism of lung cancer metastasis is urgently needed [3]. Recent studies have shown that cancer progression and metastasis are not only associated with the properties of tumor cells, but also depend on the tumor microenvironment (TME) [4]. Among the stromal cells of TME, cancer-associated fibroblasts (CAFs) are a dominant component, exhibiting a classic spindle-shape morphology with a potential for planar polarity and over-expression of marker proteins including alpha-smooth muscle actin (α-SMA) and fibroblast-activated protein (FAP) [5,6]. Accumulating research has demonstrated that cancer cells can establish cross-talk with CAFs to enhance tumor metastasis [7], and a strong association between CAFs and poor prognosis has been shown in several types of cancer, including lung cancer [[8], [9], [10]].

CAFs can secrete cytokines, growth factors, CAF-specific proteins and exosomes to support cancer cell growth, metastasis and confer chemoresistance to lung cancer [11,12]. CAFs support the growth of lung cancer cells in vivo by the secretion of the cytokines cardiotrophin-like cytokine factor 1 (CLCF1) and IL6 that directly stimulate the growth of cancer cells [13], IL-22 secreted from CAFs significantly increases the proliferation, migration and invasion of lung cancer cells but reduces apoptosis via the activation of PI3K-Akt-mTOR signaling [14]. A subset of CAFs, CD90 (+) CAFs, secrete insulin-like growth factor-II (IGF-II) activated IGF1R signaling in cancer cells, induce nanog expression and promote stemness [15]. Also, IGF-1 and hepatocyte growth factor(HGF), secreted by CAFs, synergistically induce Annexin A2(ANXA2) expression, thus promoting epithelial–mesenchymal transition (EMT) and drug resistance in non-small cell lung cancers (NSCLCs) [16]. Recent research has found that CAFs deliver SNAIL1 to recipient cells via exosomes and induce EMT in lung cancer cells [17].

To better understand the mechanism of the interaction of CAFs with lung cancer cell in growth and invasion, the current study demonstrated that CAF-conditional medium (CM) enhanced the proliferation and invasion of the recipient cancer cells. Interestingly, we found that vascular cell adhesion molecule-1 (VCAM-1), secreted from CAFs, was upregulated compared with the normal fibroblasts (NFs), and blocking the expression of VCAM-1 in CAFs led to the attenuation of the proliferation and invasion of cancer cells. Moreover, we verified that VCAM-1 secreted from CAFs activated the AKT and MAPK pathway via a receptor, the α4β1 integrin (also known as very-late antigen, VLA-4) in cancer cells. In vivo, we proved that VCAM-1 increased tumor growth. Collectively, the present work provides solid evidence that VCAM-1, secreted from CAFs, enhances the growth and invasion of lung cancer cells, implying that VCAM-1 may be a potential anti-metastasis therapeutic target of lung cancer.

Section snippets

Cell culture and reagents

A549 and H358 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). A549 was cultured in DMEM/F12 (Gibco BRL, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (FBS, Hyclone, Logan, UT, USA), and H358 cells were cultured in RPMI 1640 (Gibco BRL) supplemented with 10% FBS at 37 °C in a 5% CO₂ incubator. Recombinant human VCAM-1(150-04-100) was obtained from Peprotech (Rocky Hill, NJ, USA). VCAM-1(AF809) and Sheep IgG (5-001-A) were purchased from

The conditional medium (CM) of CAFs promotes the proliferation, migration and invasion of recipient lung cancer cells

Primary cells were isolated from lung cancer and adjacent normal tissues of three patients. Consistently with previous reports [3], the morphology of the two primary cells is a long-spindle shape, suggesting that these cells were fibroblasts (Fig. 1A). Western blotting analysis results indicated that more α-SMA and FAP were expressed in CAFs than NFs in three paired primary cultured cells (Fig. 1B). Meanwhile, α-SMA, FAP and PDFGR-α of CAFs were higher than two generation NFs (NFs-P2), and the

Discussion

Numerous evidence has demonstrated that CAFs play a critical role in cancer procession, including altering the immune response and inflammation, maintaining stemness and metabolism, inhibiting apoptosis, and enhancing proliferation, invasion and metastasis, and so on [29,30]. Although some studies have found that CAFs are involved in the growth and metastasis of lung cancer [3,14,31], the role of CAFs in lung cancer and its mechanism have still not been fully elucidated. Here, we further

Declaration of competing interest

The authors declare no conflict of interest.

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant Nos. 81672761 and 81372182) and the Natural Science Foundation of Hunan Province (2018JJ2545).

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Citation Excerpt :
In addition, apCAFs express MHC II and CD74, which contribute to antigen presentation and immune modulation functions [13,35]. CAFs are highly heterogeneous stromal cells, and their effects on tumor cells are mediated by a complex and complicated signaling pathways that involve transforming growth factor-beta (TGF-β) [36,37], MAPK [38,39], PI3K/AKT/mTOR [40,41], JAK/STAT [42,43], Wnt/β-catenin [44], NF-κB [45], and integrin β1-FAK-Src [46,47]. All of these signaling networks of CAF influence tumor cells and have enormous potential as targets for impeding CAF-mediated tumor promotion.
Current cancer therapeutic strategies are generally not sufficient to eradicate malignancy, as cancer stroma cells contribute to tumor evasion and therapeutic resistance. Cancer-associated fibroblasts (CAFs) constitute a largely heterogeneous type of stromal cell population and are important components of the tumor microenvironment (TME). CAFs are the most abundant stromal cell type and are actively involved in tumor progression through complex mechanisms involving effects on other cell types. Research conducted in recent years has emphasized an emerging function of CAFs in the remodeling of the TME that promotes tumor progression with effects on response to treatment by various molecular mechanisms. A comprehensive mechanism of tumor-promoting activities of CAFs could facilitate the development of novel diagnostic and therapeutic approaches. In this review, the biological characterization of CAFs and the mechanisms of their effects on TME remodeling are summarized. Furthermore, we also highlight currently available therapeutic strategies targeting CAF in the context of optimizing the success of immunotherapies and briefly discuss possible future perspectives and challenges related to CAF studies.

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¹: These authors contributed equally to this work.

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Original ArticlesVCAM-1 secreted from cancer-associated fibroblasts enhances the growth and invasion of lung cancer cells through AKT and MAPK signaling

Highlights

Abstract

Introduction

Section snippets

Cell culture and reagents

The conditional medium (CM) of CAFs promotes the proliferation, migration and invasion of recipient lung cancer cells

Discussion

Declaration of competing interest

Acknowledgements

Lung Cancer

Adv. Drug Deliv. Rev.

Semin. Cancer Biol.

J. Thorac. Oncol.

Biochim. Biophys. Acta (BBA) - Mol. Basis Dis.

Cancer Lett.

Cancer Cell

Cancer Cell

Cancer Lett.

Cancer Lett.

Mol. Cancer

J. Thorac. Oncol.

Biochim. Biophys. Acta (BBA) - Mol. Basis Dis.

Blood

Cancer statistics

CA A Cancer J. Clin.

Cancer-associated fibroblasts enhance metastatic potential of lung cancer cells through IL-6/STAT3 signaling pathway

Oncotarget

Microenvironmental regulation of tumor progression and metastasis

Nat. Med.

The biology and function of fibroblasts in cancer

Nat. Rev. Cancer

The roles of tumor- and metastasis-promoting carcinoma-associated fibroblasts in human carcinomas

Cell Tissue Res.

mir-1-mediated paracrine effect of cancer-associated fibroblasts on lung cancer cell proliferation and chemoresistance

Oncol. Rep.

Original Articles
VCAM-1 secreted from cancer-associated fibroblasts enhances the growth and invasion of lung cancer cells through AKT and MAPK signaling