Dataset Information

Bladder cancer cells re-educate TAMs through lactate shuttling in the microfluidic cancer microenvironment.

ABSTRACT:

Background

In the present study, we aimed to investigate the influence of lactate shuttling on the functional polarization and spatial distribution of transitional cell carcinoma of the bladder (TCCB) cells and macrophages.

Methods

We designed a microfluidic coculture chip for real-time integrative assays. The effect of lactate shuttling on the re-education of macrophages by TCCB cells was explored by measuring the levels of NO using a total NO assay kit and by evaluating the protein expression of iNOS, p-NFkB-p65, Arg-1 and HIF-1? via cell immunofluorescence and western blotting. Additionally, we examined TCCB cell viability using acridine orange/ethidium bromide (AO/EB) and MitoTracker staining. Moreover, the concentration distributions of lactate and large signaling proteins in the culture chambers were measured using 4',6-diamidino-2-phenylindole (DAPI) and fluorescein isothiocyanate-dextran (FITC-dextran). Furthermore, the recruitment of macrophages and the influence of macrophages on BC metastasis were observed via light microscopy.

Results

We confirmed that TCCB cells reprogrammed macrophages into an M2 phenotype. Moreover, lactate inhibited M1 polarization and induced M2 polarization of macrophages, but blockade of cancer cell-macrophage lactate flux significantly inhibited the re-education of macrophages by TCCB cells. In addition, lactate diffused faster and deeper than large signaling proteins in the microfluidic tumor microenvironment. Furthermore, lactate alone induced the migration of macrophages, and M1, but not M2, macrophages reduced the motility of TCCB cells.

Conclusions

TCCB cells reprogrammed macrophages into an M2 phenotype in a manner that depended on cancer cell-TAM lactate flux. Furthermore, the lactate shuttle may be a determinant of the density of TAMs in tumor tissue.

SUBMITTER: Zhao Y

PROVIDER: S-EPMC4770766 | biostudies-literature | 2015 Nov

REPOSITORIES: biostudies-literature

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Publications

Bladder cancer cells re-educate TAMs through lactate shuttling in the microfluidic cancer microenvironment.

Zhao Yang Y Wang Degui D Xu Ting T Liu Pengfei P Cao Yanwei Y Wang Yonghua Y Yang Xuecheng X Xu Xiaodong X Wang Xinsheng X Niu Haitao H

Oncotarget 20151101 36

<h4>Background</h4>In the present study, we aimed to investigate the influence of lactate shuttling on the functional polarization and spatial distribution of transitional cell carcinoma of the bladder (TCCB) cells and macrophages.<h4>Methods</h4>We designed a microfluidic coculture chip for real-time integrative assays. The effect of lactate shuttling on the re-education of macrophages by TCCB cells was explored by measuring the levels of NO using a total NO assay kit and by evaluating the prot ...[more]

PMID: 26474279

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Dataset Information

Bladder cancer cells re-educate TAMs through lactate shuttling in the microfluidic cancer microenvironment.

Background

Methods

Results

Conclusions

Publications

Bladder cancer cells re-educate TAMs through lactate shuttling in the microfluidic cancer microenvironment.

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