Artesunate and chloroquine induce cytotoxic activity on cholangiocarcinoma cells via different cell death mechanisms

Diwakar Guragain,Chaisiri Wongkham,Kulthida Vaeteewoottacharn,Ubon Cha'On,Wunchana Seubwai,Daiki Kobayashi,Sopit Wongkham,Atit Silsinivanit,Kanlayanee Sawanyawisuth,Norie Araki

doi:10.14715/cmb/2018.64.10.18

Diwakar Guragain, Chaisiri Wongkham + Show 8 more

Open Access

https://doi.org/10.14715/cmb/2018.64.10.18

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Abstract

Chemotherapy for cholangiocarcinoma (CCA) is not quite successful. In this study, we revisited the possibility of artesunate (ART) and chloroquine (CQ), the antimalarial drugs, as therapeutic agents against CCA. The possible mechanisms of these drugs to exert cytotoxicity on CCA cells were also explored. The effects of ART and CQ on proliferation and death patterns of two CCA cell lines, KKU-214 and its highly metastatic subtype KKU-214L5, were examined using water soluble tetrazolium (WST) assay and time-lapse photometry, respectively. To differentiate and verify the death patterns between necrosis and apoptosis, lactate dehydrogenase (LDH) release, and caspase 3 activity were measured. CellROXTM green reagent staining method was used to assess reactive oxygen species (ROS) production in ART- and CQ-treated cells. ART and CQ significantly inhibited proliferation of CCA cells. Both drugs kill malarial parasites via similar mechanism depending on ROS formation, however, ART induced necrotic cell death and CQ induced apoptotic cell death in CCA cells. ART induced LDH release, whereas CQ activated caspase 3, confirming induction of necrotic and apoptotic cell deaths by ART and CQ, respectively. ART treatment induced higher ROS production than CQ. ART and CQ induce CCA cells death via different death pathways. ART should be suitable for necrosis-sensitive CCA, whereas CQ is more suitable for apoptosis-sensitive CCA.

Highlights

Cholangiocarcinoma (CCA) is a malignancy of bile duct epithelial cells
ART should be suitable for necrosis-sensitive CCA, whereas CQ is more suitable for apoptosis-sensitive CCA
When cells were exposed to various doses of ART (0 - 340 μM) and CQ (0 - 50 μM) for 24, 48, and 72 h, both CQ and ART inhibited the proliferation of CCA cells in a dose- and time-dependent manner (Figure 1)

Summary

Introduction

The incidence of CCA is extremely high in the Greater Mekong Subregion (GMS), in the Northeast Thailand, whereas it is very low in other parts of the world [1]. The problems of CCA are the silent clinical presentation and the difficulty of diagnosis at the early stage. The radical resection is the only effective treatment but almost all CCA-patients come to hospitals with advanced and incurable stage of the disease [3, 4]. Currently patients are prescribed traditional chemotherapy drugs, a new regimen is being looked for. A new indication for existing drugs [6, 7], may be one of the strategy to improve clinical outcomes of chemotherapy. The main advantage of drug repurposing is, as the toxicity profile of the drugs have already been known, and the chances of drug failure due to toxicity can be minimized

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