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Encapsulation in PLGA-PEG enhances 9- nitro-camptothecin cytotoxicity to human ovarian carcinoma cell line through apoptosis pathway.

Ahmadi F, Derakhshandeh K, Jalalizadeh A, Mostafaie A, Hosseinzadeh L - Res Pharm Sci (2015 Mar-Apr)

Bottom Line: The results showed that activation of caspase-3 and -9 significantly increased by free 9-NC and PLGA-PEG loaded nanoparticles in A2780 cells.In contrast to the free drug which increased the activation of caspase-8, 9-NC-loaded PLGA-PEG nanoparticles did not alter the activation of caspase-8.Moreover, our results confirmed that 9-NC in nanoparticles at the level of gene expression potentiated down-regulation of Bcl-2, up regulation of Bax, and Smac/DIABLO leading to a decrease in mitochondrial membrane potential.

View Article: PubMed Central - PubMed

Affiliation: Nano Drug Delivery Research Center, School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, I.R. Iran.

ABSTRACT
Ovarian cancer is the fifth leading cause of the cancer-related death among women. 9-nitrocamptothecin (9-NC) is a water-insoluble derivative of camptothecin used for the treatment of patients with advanced ovarian cancer. Previous studies showed that the encapsulation of 9-NC in poly (lactic-co-glycolic acid, PLGA) nanoparticles increased the cytotoxic effect of the drug on different cancer cell lines. In the present study, the cytotoxic effects of 9-NC, 9-NC-loaded PLGA and PLGA-polyethylene glycol (PLGA-PEG) nanoparticles with varying degree of PEG (5, 10, and 15%) were evaluated on human ovarian carcinoma cell line. Furthermore, the mode of cell death induced by 9-NC and the optimized 9-NC-loaded PLGA-PEG nanoparticles on A2780 cell line were investigated. 9-NC incorporating nanoparticles were prepared by nanopercipitation method and their physicochemical characteristics were evaluated using standard methods. The results showed that activation of caspase-3 and -9 significantly increased by free 9-NC and PLGA-PEG loaded nanoparticles in A2780 cells. In contrast to the free drug which increased the activation of caspase-8, 9-NC-loaded PLGA-PEG nanoparticles did not alter the activation of caspase-8. Collectively, it appears that apoptosis induced by 9-NC incorporated in PLGA-PEG 5% occurred through the activation of caspase-9 rather than activation of caspase-8 which is the mediator of extrinsic pathway. Moreover, our results confirmed that 9-NC in nanoparticles at the level of gene expression potentiated down-regulation of Bcl-2, up regulation of Bax, and Smac/DIABLO leading to a decrease in mitochondrial membrane potential. Taken together, our results showed that 9-NC incorporated in PLGA-PEG 5% nanoparticles is able to induce apoptosis in A2780 human ovarian carcinoma cells and has the potential for the treatment of ovarian carcinoma.

No MeSH data available.


Related in: MedlinePlus

The effect of 9-NC and 9 NC-PLGA-PEG 5% (9-NC-NP) on A; Bax, B; Bcl-2 and Smac/DIABLO mRNA expression and C; Bax/Bcl2 in A2780 human cancer cells. Normalization relative to b-actin was performed. Levels of mRNA are expressed relative to control in the mean ± S.E.M values derived from three independent experiments.*; P<0.05 and ***; P<0.001 vs. control.
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Figure 4: The effect of 9-NC and 9 NC-PLGA-PEG 5% (9-NC-NP) on A; Bax, B; Bcl-2 and Smac/DIABLO mRNA expression and C; Bax/Bcl2 in A2780 human cancer cells. Normalization relative to b-actin was performed. Levels of mRNA are expressed relative to control in the mean ± S.E.M values derived from three independent experiments.*; P<0.05 and ***; P<0.001 vs. control.

Mentions: Real-time RT-PCR analysis clearly showed a significant reduction in the expression level a significant reduction in the expression level of Bcl-2 after 24 h treatment with 9-NC-loaded PLGA-PEG 5% nanoparticles. Moreover, induction of apoptosis by 9-NC -PLGA-PEG 5% was accompanied by increase in mRNA expression of Bax and Smac/DIABLO genes. Surprisingly, we did not show any significant change in the mRNA expression of Bcl-2 and Smac/DIABLO genes upon treatment with 9-NC. Induction of apoptosis by 9-NC was only accompanied by increase in mRNA levels of proapoptotic Bax in A2780 cells (Fig. 4a–c).


Encapsulation in PLGA-PEG enhances 9- nitro-camptothecin cytotoxicity to human ovarian carcinoma cell line through apoptosis pathway.

Ahmadi F, Derakhshandeh K, Jalalizadeh A, Mostafaie A, Hosseinzadeh L - Res Pharm Sci (2015 Mar-Apr)

The effect of 9-NC and 9 NC-PLGA-PEG 5% (9-NC-NP) on A; Bax, B; Bcl-2 and Smac/DIABLO mRNA expression and C; Bax/Bcl2 in A2780 human cancer cells. Normalization relative to b-actin was performed. Levels of mRNA are expressed relative to control in the mean ± S.E.M values derived from three independent experiments.*; P<0.05 and ***; P<0.001 vs. control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4584455&req=5

Figure 4: The effect of 9-NC and 9 NC-PLGA-PEG 5% (9-NC-NP) on A; Bax, B; Bcl-2 and Smac/DIABLO mRNA expression and C; Bax/Bcl2 in A2780 human cancer cells. Normalization relative to b-actin was performed. Levels of mRNA are expressed relative to control in the mean ± S.E.M values derived from three independent experiments.*; P<0.05 and ***; P<0.001 vs. control.
Mentions: Real-time RT-PCR analysis clearly showed a significant reduction in the expression level a significant reduction in the expression level of Bcl-2 after 24 h treatment with 9-NC-loaded PLGA-PEG 5% nanoparticles. Moreover, induction of apoptosis by 9-NC -PLGA-PEG 5% was accompanied by increase in mRNA expression of Bax and Smac/DIABLO genes. Surprisingly, we did not show any significant change in the mRNA expression of Bcl-2 and Smac/DIABLO genes upon treatment with 9-NC. Induction of apoptosis by 9-NC was only accompanied by increase in mRNA levels of proapoptotic Bax in A2780 cells (Fig. 4a–c).

Bottom Line: The results showed that activation of caspase-3 and -9 significantly increased by free 9-NC and PLGA-PEG loaded nanoparticles in A2780 cells.In contrast to the free drug which increased the activation of caspase-8, 9-NC-loaded PLGA-PEG nanoparticles did not alter the activation of caspase-8.Moreover, our results confirmed that 9-NC in nanoparticles at the level of gene expression potentiated down-regulation of Bcl-2, up regulation of Bax, and Smac/DIABLO leading to a decrease in mitochondrial membrane potential.

View Article: PubMed Central - PubMed

Affiliation: Nano Drug Delivery Research Center, School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, I.R. Iran.

ABSTRACT
Ovarian cancer is the fifth leading cause of the cancer-related death among women. 9-nitrocamptothecin (9-NC) is a water-insoluble derivative of camptothecin used for the treatment of patients with advanced ovarian cancer. Previous studies showed that the encapsulation of 9-NC in poly (lactic-co-glycolic acid, PLGA) nanoparticles increased the cytotoxic effect of the drug on different cancer cell lines. In the present study, the cytotoxic effects of 9-NC, 9-NC-loaded PLGA and PLGA-polyethylene glycol (PLGA-PEG) nanoparticles with varying degree of PEG (5, 10, and 15%) were evaluated on human ovarian carcinoma cell line. Furthermore, the mode of cell death induced by 9-NC and the optimized 9-NC-loaded PLGA-PEG nanoparticles on A2780 cell line were investigated. 9-NC incorporating nanoparticles were prepared by nanopercipitation method and their physicochemical characteristics were evaluated using standard methods. The results showed that activation of caspase-3 and -9 significantly increased by free 9-NC and PLGA-PEG loaded nanoparticles in A2780 cells. In contrast to the free drug which increased the activation of caspase-8, 9-NC-loaded PLGA-PEG nanoparticles did not alter the activation of caspase-8. Collectively, it appears that apoptosis induced by 9-NC incorporated in PLGA-PEG 5% occurred through the activation of caspase-9 rather than activation of caspase-8 which is the mediator of extrinsic pathway. Moreover, our results confirmed that 9-NC in nanoparticles at the level of gene expression potentiated down-regulation of Bcl-2, up regulation of Bax, and Smac/DIABLO leading to a decrease in mitochondrial membrane potential. Taken together, our results showed that 9-NC incorporated in PLGA-PEG 5% nanoparticles is able to induce apoptosis in A2780 human ovarian carcinoma cells and has the potential for the treatment of ovarian carcinoma.

No MeSH data available.


Related in: MedlinePlus