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Non–small-cell lung cancer (NSCLC) accountsfor the most cancer deaths worldwide, alongwith a 5-year survival of only 5% to 15%.Chemotherapy remains an important modalityfor treatment of this tumour. However, responserates to chemotherapy are only approximately30%, making its treatment a challenge. Themajor limitations include short drug half-lives,insufficient delivery, and suboptimal specificityfor malignant tissue.

There has been considerable interest in the useof stem cells as trophic vehicles for delivery ofdrugs, proteins, and other therapeutic agentsspecifically to tumours due to their lack ofimmune rejection and inherent tumour-homingcapabilities.

We hypothesized human adipose-derivedmesenchymal stem cells (hAD-MSCs) primed invitro with anti-cancer drugs could release drugsand inhibit tumour growth, thereby serving asvehicles for delivering effective and targetedtherapy to tumours.

Introduction

Experimental Model

Materials and Methods

hAD-MSCs were isolated from peri-umbilicaladipose tissue and stably transfected withgreen fluorescent protein (GFP); three humannon-small-cell lung cancer cell lines (A549,HCC827 and H520) were cultured .

Doxorubicin internalisation was analysed byfluorescence microscopy.

Cell proliferation was detected by Alarma blueassay. Cell migration was assessed by Boydenchamber assay.

Paclitaxel concentration was determined byUHD accurate-mass Q-TOF LC/MS.

Results

24 hours 48 hours 72 hours

Paclitaxel produced a strong anti-proliferative effect on three lung carcinoma cells (A549, HCC827 andH520) in a dose- and time- dependent manner whereas hAD-MSCs were highly resistant.

Drug Intake

Drug Release

hAD-MSC

Tumour cell

Chemotherapeuticdrugs

Paclitaxel (ng/ml)Paclitaxel (ng/ml)Paclitaxel (ng/ml)

After 1 hour priming, the internalisation of doxorubicin by hAD-MSCs was appreciable. The staining wasintense and enriched in cytoplasm at the end of priming (24 hours). After 24 hours, doxorubicindistribution in cytoplasm decreased, suggesting a possible excretion. Scale bar: 100 μm.

1School of Healthcare Science, Faculty of Science and Engineering, Manchester Metropolitan University2School of Science & The Environment, Faculty of Science and Engineering, Manchester Metropolitan University

3School of Biomedicine, Faculty of Medicine and Human Sciences, University of Manchester

Wen-Hui Fang1, David Smith2, Shant Kumar1, 3, Glenn Ferris1, Garry McDowell1, Mark Slevin1

Conclusion and Future Work

hAD-MSCs primed in vitro with chemotherapeu-

tic drugs could release drugs and inhibit lung

cancer cell growth, and the tropism of hAD-

MSCs to human lung carcinomas might be

exploited to therapeutic advantage as vehicles

for delivering targeted therapy to tumours.

Future work would focus on mechanisms of

chemotherapeutic drug intake and release by

hAD-MSCs, the enhanced and more specific

tumour-homing, and the utilization of

therapeutic modified hAD-MSCs in the

treatment of cancer.

hAD-MSCs were exposed to 100 μg/ml paclitaxel for 24 hours. Paclitaxel primed hAD-MSCs were further

cultured and conditioned media were collected and replaced every day. The concentrations of paclitaxel

in conditioned media were measured by LC-MS/MS and conditioned media were tested for anti-

proliferative activity on A549 and HCC827 cells.

Wild type hAD-MSCs

Day 2 Day 3 Day 7 Day 21

DAPI

DOX

Merge

GFP-hAD-MSCs

Untreated 1 hour 2 hours

DAPI

DOX

4 hours 8 hours 12 hours 24 hours

Untreated 1 hour 2 hours 4 hours 8 hours 12 hours 24 hours

This work was supported by a grant of the Ministry of National Education, CNCS – UEFISCDI, project number PN-II-ID-PCE-2012-4-0133. Many thanks to Dr Valentina Ceserani, Dr Andrew Ryan and Dr John Brognard for kindly providing cell lines.

Lund TC et al. Nat Rev Clin Oncol, 2015;12:163-74; Pessina A et al. J Control Release, 2014;192:262-70; Keung EZ et al. Stem Cells, 2013;31:227-35.

Paclitaxel-primed hAD-MSCs steadily released

paclitaxel into media over 7 days.

Drug release (Day)

Doxorubicin (DOX) 50 μM

Doxorubicin (DOX) 50 μM

GFP

Merge

The conditioned media from paclitaxel primed

hAD-MSCs produced a potent growth inhibition

on lung cancer cells.

Drug release (Day)

Chemotherapeutic Drug Primed Mesenchymal Stem Cells Induce

Anti-cancer Effects on Human Lung Cancer Cells

Chemotherapeutic Drug Primed Mesenchymal Stem Cells Induce

Anti-cancer Effects on Human Lung Cancer Cells

hAD-MSC A549 HCC827

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Positive control Conditioned media of lung cancer cells

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Media control

Acknowledgement and Key References

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Conditioned medium from human lung carcinomas

promoted hAD-MSCs migration. Scale bar: 100 μm.

4. The oncotropism of hAD-MSCs

3. Paclitaxel-primed hAD-MSCs sustainedly released paclitaxel

2. hAD-MSCs ware resistant to paclitaxel

1. The uptake and release of doxorubicin by hAD-MSCs