Time and concentration dependency of P-gp, MRP1 and MRP5 induction in response to gemcitabine uptake in Capan-2 pancreatic cancer cells

Kohan, Hamed Gilzad; Boroujerdi, Mehdi

doi:10.3109/00498254.2014.1001809

Cited by 23 publications

(21 citation statements)

References 29 publications

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“…Capan-2 pancreatic cancer cells, similar to many other type of cancerous cells, utilize various cellular mechanisms such as microparticle formation (51), and induction of efflux proteins against anticancer drugs (24). Several studies have shown that the resistance of pancreatic cancer cells to chemotherapeutics including gemcitabine, is the result of reduced cellular uptake via influx proteins (12) or their efficient removal from cells by efflux proteins (15,24).…”

Section: Discussionmentioning

confidence: 99%

“…Several studies have shown that the resistance of pancreatic cancer cells to chemotherapeutics including gemcitabine, is the result of reduced cellular uptake via influx proteins (12) or their efficient removal from cells by efflux proteins (15,24). Synthetic antioxidants such as BHA and BHT are widely used in pharmaceutical products as well as in the food industry to protect susceptible chemicals against oxidation (27,28).…”

Section: Discussionmentioning

confidence: 99%

“…Capan-2 pancreatic cancer cells, a human adenocarcinomatous adherent cell line, was chosen not only because it represents the aggressive type of pancreatic cancer (20,44,45), but also, it expresses several influx and efflux proteins including MRP1, MRP5, and P-gp (24,46). This cell line has been conventionally used as a model to study alteration in expression of the efflux proteins (44,46 with greater than 98% viability, were rinsed briefly with Ca +2 /Mg +2 free DPBS and 1-2 mL of 0.25% (w/v) trypsin-0.53 mM EDTA at room temperature.…”

Section: Experimental Protocolmentioning

confidence: 99%

“…We have shown in our previous investigation that expression of hENT1 transporters is significantly altered during development of gemcitabine resistance (24). Additionally, it has been reported that the sensitivity of pancreatic cancer cell lines to gemcitabine is significantly influenced by MRP5 (ABCC5) efflux protein (20,24) and to a lesser extent by MRP1 (ABCC1) (25) and P-gp (ABCB1) (25,26).…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, it has been reported that the sensitivity of pancreatic cancer cell lines to gemcitabine is significantly influenced by MRP5 (ABCC5) efflux protein (20,24) and to a lesser extent by MRP1 (ABCC1) (25) and P-gp (ABCB1) (25,26). Therefore, chemicals that can modulate expression of influx and efflux proteins have the potential to substantially alter therapeutic outcomes of chemotherapy.…”

Section: Introductionmentioning

confidence: 99%

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Calcitriol Reverses Induced Expression of Efflux Proteins and Potentiates Cytotoxic Activity of Gemcitabine in Capan-2 Pancreatic Cancer Cells

Kohan¹,

Sani²,

Boroujerdi³

2017

J Pharm Pharm Sci

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ABSTRACT-Purpose. Efflux and influx proteins play a major role in chemo-resistance by affecting the net cellular uptake of anti-cancer drugs. Hence, alteration of the efflux and influx protein expression may result in variations of chemotherapeutics uptake and consequently cell death rate. The present study investigated the effects of pre-treatment of capan-2 pancreatic cancer cells with calcitriol, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA) or silibinin on the induction of three major efflux proteins and the main gemcitabine influx protein. The influence of the pre-treatments on the net cellular uptake of gemcitabine, total ATPase activity, and cell death rate were also evaluated. Methods. Capan-2 pancreatic cancer cells were pre-treated for 24 h with calcitriol, BHT, BHA, or silibinin, followed by gemcitabine treatment. The concentration of gemcitabine was quantified using ultra-performance liquid chromatography (UPLC). Real-time polymerase chain reaction (RT-PCR) was utilized in order to investigate the expression of the mRNAs. The expression of the proteins was assessed using western blotting. Measurement of the ATPase activity was conducted utilizing a colorimetric method and viability of the cells was determined using a luminescent cell viability assay. Results. Protein expression studies showed that BHT, silibinin, and BHA increased expression of the efflux proteins and decreased the overall uptake of gemcitabine, whereas calcitriol significantly inhibited expression of the efflux proteins and increased gemcitabine uptake. Expression of specific mRNAs correlated reasonably well with the levels of corresponding proteins. Additionally, the expression of efflux proteins and ATPase activity were well correlated, signifying that the induced efflux proteins are functionally active. Moreover, pre-treatment with calcitriol resulted in a significant increase in cell death with gemcitabine treatment, whereas, BHA significantly reduced the cell death rate. On the other hand, pre-treatment with BHT and silibinin had no significant effect on the cell death rate. Conclusions. Pre-treatment of the pancreatic cancer cells with calcitriol significantly increased gemcitabine cellular uptake and consequently decreased cell viability after treatment with gemcitabine, whereas BHA significantly reduced gemcitabine uptake and decreased cell death rate, which were at least partially attributed to the alteration of expression of efflux and influx proteins.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Experimental Protocolmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Calcitriol Reverses Induced Expression of Efflux Proteins and Potentiates Cytotoxic Activity of Gemcitabine in Capan-2 Pancreatic Cancer Cells

Kohan¹,

Sani²,

Boroujerdi³

2017

J Pharm Pharm Sci

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Decreased drug resistance of bladder cancer using phytochemicals treatment

Cho

et al. 2020

The Kaohsiung J of Med Scie

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The aim of the study is to investigate the ability of phytochemicals to overcome the multiple drug resistance (MDR) of bladder cancer. 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay was used to evaluate the cytotoxic sensitivity of T24‐GCB cells, a GCB resistant cell line, to different phytochemicals, including capsaicin, quercetin, curcumin, and resveratrol, and their combination with gemcitabine. Western blot analysis was used to detect the expression of membranous ABCC2 and metabolic proteins, DCK, TK1, and TK2 in tumor cells. Animal models were used to confirm the treatment efficacy of phytochemicals in combination with gemcitabine to bladder cancer. The observed/expected ratio of cytotoxicity analysis revealed that capsaicin has synergistic effect with gemcitabine to T24‐GCB cells in a dose‐dependent pattern. Quercetin, curcumin, and resveratrol have additive effect with gemcitabine to T24‐GCB cells. Capsaicin and quercetin alone and combination with gemcitabine decreased the expression of ABCC2 and DCK and TKs, in T24‐GCB cells. On the contrary, resveratrol and curcumin alone and combination with gemcitabine increased the expression of ABCC2 but decreased cytoplasmic kinases simultaneously. In xenografted subcutaneous tumor model on nude mice, combination treatment of capsaicin and gemcitabine demonstrated the highest tumor suppression effect when compared to capsaicin or gemcitabine treatment alone. The MDR of bladder cancer is closely related to membranous ABCC2, cytoplasmic DCK, and TKs expression. Capsaicin owns the strongest synergistic cytotoxic effect of gemcitabine to T24‐GCB cells. This combination regimen may provide as an adjunctive treatment for overcoming MDR in bladder cancer.

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A novel mechanism governing the transcriptional regulation of ABC transporters in MDR cancer cells

Pokharel

Bebawy

2017

Drug Deliv. and Transl. Res.

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P-glycoprotein (P-gp/ABCB1) and multidrug resistance-associated protein 1 (MRP1/ABCC1) are the main drug efflux transporters associated with treatment failure in cancer. Much attention has been focused on the molecular mechanisms regulating the expression of these transporters as a viable approach for identifying novel drug targets in circumventing cancer multidrug resistance (MDR) clinically. In this paper, we examine the role of miR-326 in the context of its intercellular transfer between cancer cells by extracellular membrane vesicles called microparticles (MPs). We observe that cellular suppression of ABCC1 by miR-326 is modulated by the presence of ABCB1 transcript. Specifically, we show that siRNA silencing of MP-transferred ABCB1 transcript reverses the knockdown effects of miRNA-326 on target MRP1/ABCC1 transcripts. We also demonstrate a dominance of ABCB1 transcripts when co-localized with ABCC1 transcripts, which is consistent with the facilitation of miR-326 function by ABCB1. This study identifies a novel pathway regulating the expression of ABC transporters and positions ABCB1 mRNA as a transcriptional regulator of other members of this superfamily in multidrug resistant cells through its actions on miRNAs.

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Time and concentration dependency of P-gp, MRP1 and MRP5 induction in response to gemcitabine uptake in Capan-2 pancreatic cancer cells

Cited by 23 publications

References 29 publications

Calcitriol Reverses Induced Expression of Efflux Proteins and Potentiates Cytotoxic Activity of Gemcitabine in Capan-2 Pancreatic Cancer Cells

Calcitriol Reverses Induced Expression of Efflux Proteins and Potentiates Cytotoxic Activity of Gemcitabine in Capan-2 Pancreatic Cancer Cells

Decreased drug resistance of bladder cancer using phytochemicals treatment

A novel mechanism governing the transcriptional regulation of ABC transporters in MDR cancer cells

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