Comparison of the novel HK-2 human renal proximal tubular cell line with the standard LLC-PK1 cell line in studying drug-induced nephrotoxicity

Gunness, Patrina; Aleksa, Katarina; Kosuge, Kazuhiro; Itô, Shinya; Koren, Gideon

doi:10.1139/y10-023

Cited by 76 publications

(51 citation statements)

References 35 publications

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“…We selected Human Kidney 2 (HK-2) cells as a model as these cells are an adult, noncancerous, immortalized human epithelial cell line. HK-2 cells maintain biochemical properties and activity similar to in vivo proximal tubule cells [14,15,16]. Additionally, HK-2 cells express OAT1 and OAT3 proteins [17], which facilitate TFV transport into proximal tubule cells.…”

Section: Introductionmentioning

confidence: 99%

Establishment of HK-2 Cells as a Relevant Model to Study Tenofovir-Induced Cytotoxicity

Murphy

Stafford

Petrasovits

et al. 2017

IJMS

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Tenofovir (TFV) is an antiviral drug approved for treating Human Immunodeficiency Virus (HIV) and Hepatitis B. TFV is administered orally as the prodrug tenofovir disoproxil fumarate (TDF) which then is deesterified to the active drug TFV. TFV induces nephrotoxicity characterized by renal failure and Fanconi Syndrome. The mechanism of this toxicity remains unknown due to limited experimental models. This study investigated the cellular mechanism of cytotoxicity using a human renal proximal tubular epithelial cell line (HK-2). HK-2 cells were grown for 48 h followed by 24 to 72 h exposure to 0–28.8 μM TFV or vehicle, phosphate buffered saline (PBS). MTT (MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) and Trypan blue indicated that TFV diminished cell viability at 24–72 h. TFV decreased ATP levels at 72 h when compared to vehicle, reflecting mitochondrial dysfunction. TFV increased the oxidative stress biomarkers of protein carbonylation and 4-hydroxynonenol (4-HNE) adduct formation. Tumor necrosis factor alpha (TNFα) was released into the media following exposure to 14.5 and 28.8 μM TFV. Caspase 3 and 9 cleavage was induced by TFV compared to vehicle at 72 h. These studies show that HK-2 cells are a sensitive model for TFV cytotoxicity and suggest that mitochondrial stress and apoptosis occur in HK-2 cells treated with TFV.

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Section: Introductionmentioning

confidence: 99%

Establishment of HK-2 Cells as a Relevant Model to Study Tenofovir-Induced Cytotoxicity

Murphy

Stafford

Petrasovits

et al. 2017

IJMS

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“…Mesangial cells are perivascular pericytes located within the central portion of the glomerular tuft between the capillary loops and are involved in the control of glomerular hemodynamics [26-28]. HK-2 cells retain the functional characteristics of isolated proximal tubular cells and represent an alternative to primary cultures of human cells [29-31]. The three metal oxides used in this study are relevant NPs types (produced in high tonnage) and are in widespread use in a number of consumer products.…”

Section: Introductionmentioning

confidence: 99%

Cytotoxicity and oxidative stress induced by different metallic nanoparticles on human kidney cells

et al. 2011

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BackgroundSome manufactured nanoparticles are metal-based and have a wide variety of applications in electronic, engineering and medicine. Until now, many studies have described the potential toxicity of NPs on pulmonary target, while little attention has been paid to kidney which is considered to be a secondary target organ. The objective of this study, on human renal culture cells, was to assess the toxicity profile of metallic nanoparticles (TiO2, ZnO and CdS) usable in industrial production. Comparative studies were conducted, to identify whether particle properties impact cytotoxicity by altering the intracellular oxidative status.ResultsNanoparticles were first characterized by size, surface charge, dispersion and solubility. Cytotoxicity of NPs was then evaluated in IP15 (glomerular mesangial) and HK-2 (epithelial proximal) cell lines. ZnO and CdS NPs significantly increased the cell mortality, in a dose-dependent manner. Cytotoxic effects were correlated with the physicochemical properties of NPs tested and the cell type used. Analysis of reactive oxygen species and intracellular levels of reduced and oxidized glutathione revealed that particles induced stress according to their composition, size and solubility. Protein involved in oxidative stress such as NF-κb was activated with ZnO and CdS nanoparticles. Such effects were not observed with TiO2 nanoparticles.ConclusionOn glomerular and tubular human renal cells, ZnO and CdS nanoparticles exerted cytotoxic effects that were correlated with metal composition, particle scale and metal solubility. ROS production and oxidative stress induction clearly indicated their nephrotoxic potential.

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“…For example, the HK-2 cell line generated from a culture of primary proximal tubule cells taken from normal adult cortex [28] has been extensively used as a model of drug-induced nephrotoxicity [11]. Given that cellular exposure to nephrotoxins is dependent upon the expression levels and substrate specificities of both uptake and efflux transporters, it is surprising that transporter expression in HK-2 cells has not been fully characterised.…”

Section: Introductionmentioning

confidence: 99%

The limitations of renal epithelial cell line HK-2 as a model of drug transporter expression and function in the proximal tubule

Jenkinson¹,

Chung²,

Loon³

et al. 2012

Pflugers Arch - Eur J Physiol

163

108

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Acquiring a mechanistic understanding of the processes underlying the renal clearance of drug molecules in man has been hampered by a lack of robust in vitro models of human proximal tubules. Several human renal epithelial cell lines derived from the renal cortex are available, but few have been characterised in detail in terms of transporter expression. This includes the HK-2 proximal tubule cell line, which has been used extensively as a model of nephrotoxicity. The aim of this study was to investigate the expression and function of drug transporters in HK-2 cells and their suitability as an in vitro model of the human proximal tubule. qPCR showed no mRNA expression of the SLC22 transporter family (OAT1, OAT3, OCT2) in HK-2 cells compared to renal cortex samples. In contrast, SLC16A1 (MCT1), which is important in the uptake of monocarboxylates, and SLCO4C1 (OATP4C1) were expressed in HK-2 cells. The functional expression of these transporters was confirmed by uptake studies using radiolabelled prototypic substrates DL-lactate and digoxin, respectively. The mRNA expression of apical membrane efflux transporters ABCB1 (MDR1) and several members of the ABCC family (multidrug resistance proteins, MRPs) was shown by qPCR. ABCG1 (BCRP) was not detected. The efflux of Hoechst 33342, a substrate for MDR1, was blocked by MDR1 inhibitor cyclosporin A, suggesting the functional expression of this transporter. Similarly, the efflux of the MRP-specific fluorescent dye glutathione methylfluorescein was inhibited by the MRP inhibitor MK571. Taken together, the results of this study suggest that HK-2 cells are of limited value as an in vitro model of drug transporter expression in the human proximal tubule.

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Comparison of the novel HK-2 human renal proximal tubular cell line with the standard LLC-PK1 cell line in studying drug-induced nephrotoxicity

Cited by 76 publications

References 35 publications

Establishment of HK-2 Cells as a Relevant Model to Study Tenofovir-Induced Cytotoxicity

Establishment of HK-2 Cells as a Relevant Model to Study Tenofovir-Induced Cytotoxicity

Cytotoxicity and oxidative stress induced by different metallic nanoparticles on human kidney cells

The limitations of renal epithelial cell line HK-2 as a model of drug transporter expression and function in the proximal tubule

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