Cannabis (Cannabis sativa)
Cannabidiol changes P-gp and BCRP expression in trophoblast cell lines. CSV JSON

Objectives.Marijuana is the most commonly used illicit drug during pregnancy. Due

to high lipophilicity, cannabinoids can easily penetrate physiological barriers like

the human placenta and jeopardize the developing fetus.We evaluated the impact

of cannabidiol (CBD), a major non-psychoactive cannabinoid, on P-glycoprotein

(P-gp) and Breast Cancer Resistance Protein (BCRP) expression, and P-gp function

in a placental model, BeWo and Jar choriocarcinoma cell lines (using P-gp induced

MCF7 cells (MCF7/P-gp) for comparison).

Study design. Following the establishment of the basal expression of these transporters

in the membrane fraction of all three cell lines, P-gp and BCRP protein

and mRNA levels were determined following chronic (24–72 h) exposure to CBD,

by Western Blot and qPCR. CBD impact on P-gp efflux function was examined

by uptake of specific P-gp fluorescent substrates (calcein-AM, DiOC2(3) and rhodamine123(

rh123)). Cyclosporine A (CsA) served as a positive control.

Results. Chronic exposure to CBD resulted in significant changes in the protein and

mRNA levels of both transporters. While P-gp was down-regulated, BCRP levels

were up-regulated in the choriocarcinoma cell lines. CBD had a remarkably different

influence on P-gp and BCRP expression in MCF7/P-gp cells, demonstrating that

these are cell type specific effects. P-gp dependent efflux (of calcein, DiOC2(3) and

rh123) was inhibited upon short-termexposure to CBD.

Conclusions. Our study shows that CBD might alter P-gp and BCRP expression in

the human placenta, and inhibit P-gp efflux function.We conclude that marijuana

use during pregnancy may reduce placental protective functions and change its

morphological and physiological characteristics.

Cannabis (Cannabis sativa)

24058883

1 . Inhibition of P-gp by Cannabidiol 10µM (id=NPDI-cqaAMA)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used —

N/A

  • P-gp (ABCB1)

Cell system BeWo

Results

the accumulation measurement is intracellular accumulation as measured by fluorescence

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Assay Conditions

60 min

7.4

N/A

10 µM

2 . Inhibition of P-gp by Cannabidiol 25 µM (id=NPDI-oZGcUg)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used —

N/A

calcein-am

cannabidiol

  • P-gp (ABCB1)

Cell system BeWo

Results

the accumulation measurement is intracellular accumulation as measured by fluorescence

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Assay Conditions

60 min

7.4

25 µM

3 . Inhibition of P-gp by Cannabidiol 10 µM (id=NPDI-IoxesQ)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used —

N/A

calcein-am

cannabidiol

  • P-gp (ABCB1)

Cell system BeWo

Results

the accumulation measurement is intracellular accumulation as measured by fluorescence

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Assay Conditions

60 min

7.4

10 µM

4 . Inhibition of P-gp by Cannabidiol 25 µM (id=NPDI-mLsAbQ)

In Vitro Transporter Inhibition Experiment

Inhibition was detected.  Cutoff used —

N/A

rhodamine 123

cannabidiol

  • P-gp (ABCB1)

Cell system MCF7/P-gp

Results

the accumulation measurement is intracellular accumulation as measured by fluorescence

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Assay Conditions

60 min

7.4

25 µM

1 . Induction (mRNA) of BCRP transporter with Cannabidiol (id=NPDI-jHdxpA)

In Vitro Transporter Induction Experiment

Induction was detected.  Cutoff used —

N/A

cannabidiol

  • BCRP (ABCG2)

Cell system BeWo

Results

figure 5

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

mRNA expression

Cannabis

24h, 48h, 72h

qRT-PCR, 2􀀀1􀀀1CT method

2 . Induction of BCRP transporter (mRNA) by Cannabidiol (id=NPDI-spUvqw)

In Vitro Transporter Induction Experiment

Induction was detected.  Cutoff used —

N/A

cannabidiol

  • BCRP (ABCG2)

Cell system Jar

Results

figure 5

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

mRNA expression

Cannabis

24h, 48h, 72h

qRT-PCR, 2 CT method

3 . Induction of P-gp transporter (protein) by Cannabidiol (id=NPDI-mbNW9A)

In Vitro Transporter Induction Experiment

Induction was detected.  Cutoff used —

N/A

cannabidiol

  • P-gp (ABCB1)

Cell system MCF7/P-gp

Results

figure 4

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Protein expression

10 µM and 15 µM

Cannabis

48h, 72h

computerized image analysis system

4 . Induction of BCRP transporter (protein) by Cannabidiol (id=NPDI-olqNRw)

In Vitro Transporter Induction Experiment

Induction was detected.  Cutoff used —

N/A

cannabidiol

  • BCRP (ABCG2)

Cell system Jar

Results

figure 3

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Protein expression

10 µM

Cannabis

48h, 72h

computerized image analysis system

5 . Induction of BCRP transporter (protein) by Cannabidiol (id=NPDI-Ix-KKg)

In Vitro Transporter Induction Experiment

Induction was detected.  Cutoff used —

N/A

cannabidiol

  • BCRP (ABCG2)

Cell system Jar

Results

FIGURE 3

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Protein expression

10 µM and 15 µM

Cannabis

72h

computerized image analysis system

6 . Induction of BCRP transporter (protein) by Cannabidiol (id=NPDI-Aasqdw)

In Vitro Transporter Induction Experiment

Induction was detected.  Cutoff used —

N/A

cannabidiol

  • BCRP (ABCG2)

Cell system BeWo

Results

FIGURE 2

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Protein expression

10 µM and 15 µM

Cannabis

72h

computerized image analysis system

7 . Non-induction of BCRP transporter (protein) by Cannabiodiol (id=NPDI-PaO28A)

In Vitro Transporter Induction Experiment

Non-induction was detected.  Cutoff used —

N/A

cannabidiol

  • BCRP (ABCG2)

Cell system MCF7/P-gp

Results

figure 4

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Protein expression

Cannabis

48h, 72h

8 . Down regulation of P-gp (mRNA) by Cannabidiol (id=NPDI-E7xthw)

In Vitro Transporter Induction Experiment

Down Regulation was detected.  Cutoff used —

N/A

cannabidiol

  • P-gp (ABCB1)

Cell system Jar

Results

figure 5

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

mRNA expression

Cannabis

24h, 48h, 72h

qRT-PCR, 2 CT method

9 . Down regulation of P-gp transporter (protein) by Cannabidiol (id=NPDI-2LIXDQ)

In Vitro Transporter Induction Experiment

Down Regulation was detected.  Cutoff used —

N/A

cannabidiol

  • P-gp (ABCB1)

Cell system Jar

Results

figure 3

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Protein expression

10 µM

Cannabis

72h

computerized image analysis system

10 . Down regulation of P-gp transporter (protein) by Cannabidiol (id=NPDI-29ER6A)

In Vitro Transporter Induction Experiment

Down Regulation was detected.  Cutoff used —

N/A

cannabidiol

  • P-gp (ABCB1)

Cell system BeWo

Results

figure 2

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Protein expression

Cannabis

48h, 72h

computerized image analysis system

11 . Down regulation of P-gp transporter (protein) by Cannabidiol (id=NPDI-DZj9Zg)

In Vitro Transporter Induction Experiment

Down Regulation was detected.  Cutoff used —

N/A

cannabidiol

  • P-gp (ABCB1)

Cell system BeWo

Results

Sample Compound measured Value Measurement Study sequence Additional information N replicates

Experimental Conditions

Protein expression

10 µM and 15 µM

Cannabis

72h

computerized image analysis system