Modulation the crosstalk between tumor-associated macrophages and non-small cell lung cancer to inhibit tumor migration and invasion by ginsenoside Rh2

Abstract

BackgroundTumor-associated macrophages (TAMs) play a critical role in modulating the tumor microenvironment and promote tumor metastases. Our studies have demonstrated that ginsenoside Rh2 (G-Rh2), a monomeric compound extracted from ginseng, is a promising anti-tumor agent in lung cancer cells. However, it remains unclear whetherG-Rh2 can modulate the differentiation of TAMs and its interaction with tumor microenvironment. In this study, we investigated how G-Rh2 regulates the phenotype of macrophages and affects the migration of non-small cell lung cancer (NSCLC) cells.MethodsMurine macrophage-like RAW264.7 cells and human THP-1 monocyte were differentiated into M1 and M2 subsets of macrophages with different cytokines combination, which were further identified by flow cytometry with specific biomarkers. M2 macrophages were sorted out to co-culture with NSCLC cell lines, A549 and H1299. Wound healing assay was performed to examine the cell migration. Expression levels of matrix metalloproteinases 2 and 9 (MMP-2, − 9) and vascular endothelial growth factor-C (VEGF-C) were measured by RT-qPCR and western blot, and the release of VEGF in the supernatant was measured by a VEGF ELISA kit. Finally, modulation of TAMs phenotype and VEGF expression by G-Rh2 was examined in vivo.ResultsWe demonstrated that M2 subset of macrophages alternatively differentiated from RAW264.7 or THP-1cells promote migration of NSCLC cells. Further examinations revealed that NSCLC significantly increased the release of VEGF to the media and elevated the expression levels of VEGF at mRNA and protein levels after being co-cultured with M2 macrophages. Similar alterations in MMP-2 and MMP-9 were observed in NSCLC after being co-cultured. Of note,G-Rh2 had a potential to effectively convert M2 phenotype to M1 subset of macrophages. Importantly, G-Rh2 had a preference to decrease the expression levels of VEGF, MMP2, and MMP9 in co-cultured lung cancer cells, over than those in lung cancer cells without co-culturing. Consistently, G-Rh2 reduced M2 macrophage marker CD206 and VEGF expression levels in vivo.ConclusionsAll of these results suggested that M2 subset macrophages drive lung cancer cells with more aggressive phenotypes. G-Rh2 has a potential to convert TAMs from M2 subset to M1 in the microenvironment and prevents lung cancer cell migration, suggesting the therapeutic effects of G-Rh2onlung cancer.

Tumor-promoting effects of pancreatic cancer cell exosomes on THP-1-derived macrophages

“…All cell lines were obtained from ATCC, grown at 37°C with 5% CO 2 , and STR verified yearly by ATCC. THP-1 cells (a human monocytic cell line) represent an established in vitro model commonly used to examine TAM-tumor cells interactions [ 15 ]. AsPC-1 cells were cultured in modified RPMI 1640 containing 10% FBS, 10 mM HEPES, 1 mM sodium pyruvate, and 13.9 mM D-glucose.…”

Tumor-Associated Macrophages Secret Exosomal miR-155 to Promote Metastasis of Non-Small-Cell Lung Cancer

“…Although various biomolecules have been found to mediate lung cancer metastasis, knowledge regarding to roles of tumor microenvironment in lung cancer metastasis has largely lagged behind. Stromal cells including TAMs in the tumor microenvironment are either induced or directly interacting with cancer cells to promote the progression and metastasis of lung cancer [33,34] . The modes of action of stromal cells include the secretion of extracellular components, such as growth factors, chemokines and exosomes.…”

Tumor-associated macrophages secret exosomal miR-155 to promote metastasis of non-small-cell lung cancer

“…Stromal cells including TAMs in the tumor microenvironment are either induced or directly interacting with cancer cells to promote the progression and metastasis of lung cancer. [30,31] The modes of action of stromal cells including the secretion of extracellular components, such as growth factors, chemokines and exosomes. However, due to the extreme complexity of these processes, the molecular mechanisms underlying lung metastasis has not been fully understood.…”