Molecular and functional characterization of choline transporter in the human trophoblastic cell line JEG-3 cells

Yara, Miki; Iwao, Beniko; Hara, Naomi; Yamanaka, T.; Uchino, Haruto; Inazu, Masato

doi:10.1016/j.placenta.2015.03.010

Cited by 16 publications

(15 citation statements)

References 28 publications

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“…In contrast to our observations, Yara and colleagues [34] have recently shown expression of CTL2 in the trophoblastic cell line JEG-3 and suggest a potential role for CTL2 in the low-affinity choline transport found in these cells [26]. The discrepancy between results is likely due to the approach of analysis.…”

Section: Discussioncontrasting

confidence: 99%

Characterization of choline transporters in the human placenta over gestation

et al. 2015

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INTRODUCTION The developing fetus relies on the maternal blood supply to provide the choline it requires for making membrane lipids, synthesizing acetylcholine, and performing important methylation reactions. It is vital, therefore, that the placenta is efficient at transporting choline from maternal to fetal circulation. Although choline transporters have been found in term placenta samples, little is known about what cell types express specific choline transporters and how expression of the transporters may change over gestation. The objective of this study was to characterize choline transporter expression levels and localization in the human placenta throughout placental development. METHODS We analyzed CTL1 and −2 expression over gestation in human placental biopsies from 6 to 40 weeks gestation (n=6–10 per gestational window) by immunoblot analysis. To determine the cellular expression pattern of the choline transporters throughout gestation, immunofluorescence analysis was then performed. RESULTS Both CTL1 and CTL2 were expressed in the chorionic villi from 6 weeks gestation to term. Labor did not alter expression levels of either transporter. CTL1 localized to the syncytial trophoblasts and the endothelium of the fetal vasculature within the chorionic villous structure. CTL2 localized mainly to the stroma early in gestation and by the second trimester co-localized with CTL1 at the fetal vasculature. DISCUSSION The differential expression pattern of CTL1 and CTL2 suggests that CTL1 is the key transporter involved in choline transport from maternal circulation and both transporters are likely involved in stromal and endothelial cell choline transport.

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Section: Discussioncontrasting

confidence: 99%

Characterization of choline transporters in the human placenta over gestation

et al. 2015

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“…However, there have been some reports on this subject. CTL2 mRNA and protein are expressed in the human inner ear ( Nair et al ., 2004 ), rat renal epithelial cells ( Yabuki et al ., 2009 ), human trophoblastic cells ( Yara et al ., 2015 ), human glioblastoma cells ( Taguchi et al ., 2014 ), human brain microvascular endothelial cells ( Iwao et al ., 2016 ) and human tongue cancer cells ( Nishiyama et al ., 2016 ). The intracellular localization of CTL2 differs among these cells and it is located in the plasma membrane and mitochondria.…”

Section: Discussionmentioning

confidence: 99%

Molecular and Functional Characterization of Choline Transporter-Like Proteins in Esophageal Cancer Cells and Potential Therapeutic Targets

Nagashima

Nishiyama

Iwao

et al. 2018

Biomolecules & Therapeutics

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In this study, we examined the molecular and functional characterization of choline uptake in the human esophageal cancer cells. In addition, we examined the influence of various drugs on the transport of [3H]choline, and explored the possible correlation between the inhibition of choline uptake and apoptotic cell death. We found that both choline transporter-like protein 1 (CTL1) and CTL2 mRNAs and proteins were highly expressed in esophageal cancer cell lines (KYSE series). CTL1 and CTL2 were located in the plasma membrane and mitochondria, respectively. Choline uptake was saturable and mediated by a single transport system, which is both Na+-independent and pH-dependent. Choline uptake and cell viability were inhibited by various cationic drugs. Furthermore, a correlation analysis of the potencies of 47 drugs for the inhibition of choline uptake and cell viability showed a strong correlation. Choline uptake inhibitors and choline deficiency each inhibited cell viability and increased caspase-3/7 activity. We conclude that extracellular choline is mainly transported via a CTL1. The functional inhibition of CTL1 by cationic drugs could promote apoptotic cell death. Furthermore, CTL2 may be involved in choline uptake in mitochondria, which is the rate-limiting step in S-adenosylmethionine (SAM) synthesis and DNA methylation. Identification of this CTL1- and CTL2-mediated choline transport system provides a potential new target for esophageal cancer therapy.

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“…Subcellular localization of CTL1 and CTL2 proteins using immunocytochemistry was performed as previously described [8,9,42]. MIA PaCa-2 cells cultured on a 35 mm noncoated Glass Base dish (IWAKI, Tokyo, Japan) were fixed with 100% methanol at −20 • C for 20 min.…”

Section: Immunocytochemistrymentioning

confidence: 99%

Molecular and Functional Analysis of Choline Transporters and Antitumor Effects of Choline Transporter-Like Protein 1 Inhibitors in Human Pancreatic Cancer Cells

Hirai

Watanabe

Nishijima

et al. 2020

IJMS

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Choline, an organic cation, is one of the biofactors that play an important role in the structure and the function of biological membranes, and it is essential for the synthesis of phospholipids. Choline positron emission tomography-computed tomography (PET/CT) provides useful information for the imaging diagnosis of cancers, and increased choline accumulation has been identified in a variety of tumors. However, the molecular mechanisms of choline uptake and choline transporters in pancreatic cancer have not been elucidated. Here, we examined molecular and functional analyses of choline transporters in human pancreatic-cancer cell line MIA PaCa-2 and the elucidation of the action mechanism behind the antitumor effect of novel choline-transporter-like protein 1 (CTL1) inhibitors, Amb4269951 and its derivative Amb4269675. CTL1 and CTL2 mRNAs were highly expressed in MIA PaCa-2 cells, and CTL1 and CTL2 proteins were localized in the plasma membrane and the intracellular compartments, respectively. Choline uptake was characterized by Na+-independence, a single-uptake mechanism, and inhibition by choline-uptake inhibitor HC-3, similar to the function of CTL1. These results suggest that the uptake of extracellular choline in MIA PaCa-2 cells is mediated by CTL1. Choline deficiency and HC-3 treatment inhibited cell viability and increased caspase 3/7 activity, suggesting that the inhibition of CTL1 function, which is responsible for choline transport, leads to apoptosis-induced cell death. Both Amb4269951 and Amb4269675 inhibited choline uptake and cell viability and increased caspase-3/7 activity. Ceramide, which is increased by inhibiting choline uptake, also inhibited cell survival and increased caspase-3/7 activity. Lastly, both Amb4269951 and Amb4269675 significantly inhibited tumor growth in a mouse-xenograft model without any adverse effects such as weight loss. CTL1 is a target molecule for the treatment of pancreatic cancer, and its inhibitors Amb4269951 and Amb4269675 are novel lead compounds.

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Molecular and functional characterization of choline transporter in the human trophoblastic cell line JEG-3 cells

Cited by 16 publications

References 28 publications

Characterization of choline transporters in the human placenta over gestation

Characterization of choline transporters in the human placenta over gestation

Molecular and Functional Characterization of Choline Transporter-Like Proteins in Esophageal Cancer Cells and Potential Therapeutic Targets

Molecular and Functional Analysis of Choline Transporters and Antitumor Effects of Choline Transporter-Like Protein 1 Inhibitors in Human Pancreatic Cancer Cells

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