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Photothermal treatment of liver cancer with albumin-conjugated gold nanoparticles initiates Golgi Apparatus-ER dysfunction and caspase-3 apoptotic pathway activation by selective targeting of Gp60 receptor.

Mocan L, Matea C, Tabaran FA, Mosteanu O, Pop T, Mocan T, Iancu C - Int J Nanomedicine (2015)

Bottom Line: Darkfield microscopy and immunochemical staining was used to demonstrate the selective internalization of Alb-GNPs inside the HepG2 cells via Gp60 receptors targeting.The postirradiation apoptotic rate of HepG2 cells treated with Alb-GNPs ranged from 25.8% (for 5 μg/mL) to 48.2% (for 50 μg/mL) at 60 seconds, while at 30 minutes the necrotic rate increased from 35.7% (5 μg/mL) to 52.3% (50 μg/mL), P-value <0.001.Significantly lower necrotic rates were obtained when human hepatocytes were treated with Alb-GNPs in a similar manner.

View Article: PubMed Central - PubMed

Affiliation: Nanomedicine Department, Regional Institute of Gastroenterology and Hepatology "Octavian Fodor", University of Medicine and Pharmacy, "Iuliu Hatieganu", Croitorilor, Cluj-Napoca, Romania ; Department of Surgery, University of Medicine and Pharmacy, "Iuliu Hatieganu", Croitorilor, Cluj-Napoca, Romania.

ABSTRACT
We present a method of enhanced laser thermal ablation of HepG2 cells based on a simple gold nanoparticle (GNP) carrier system such as serum albumin (Alb), and demonstrate its selective therapeutic efficacy compared with normal hepatocyte cells. HepG2 or hepatocytes were treated with Alb-GNPs at various concentrations and various incubation times, and further irradiated using a 2 W, 808 nm laser. Darkfield microscopy and immunochemical staining was used to demonstrate the selective internalization of Alb-GNPs inside the HepG2 cells via Gp60 receptors targeting. The postirradiation apoptotic rate of HepG2 cells treated with Alb-GNPs ranged from 25.8% (for 5 μg/mL) to 48.2% (for 50 μg/mL) at 60 seconds, while at 30 minutes the necrotic rate increased from 35.7% (5 μg/mL) to 52.3% (50 μg/mL), P-value <0.001. Significantly lower necrotic rates were obtained when human hepatocytes were treated with Alb-GNPs in a similar manner. We also showed by means of immunocytochemistry that photothermal treatment of Alb-conjugated GNPs in liver cancer initiates Golgi apparatus-endoplasmic reticulum dysfunction with consequent caspase-3 apoptotic pathway activation and cellular apoptosis. The presented results may become a new method of treating cancer cells by selective therapeutic vectors using nanolocalized thermal ablation by laser heating.

No MeSH data available.


Related in: MedlinePlus

The concomitent detection of Golgi apparatus function and caspase 3 pathway.Notes: Upper row: control sample (no Alb-GNPs exposure: low red fluorescence is visible, suggesting that Hep G2 cells have low apoptotic state under normal conditions; in contrast Golgi function is normal as suggested by green fluorescence. Middle row: exposure to 10 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). Lower green fluorescence intensity staining combined with increased red fluorescence suggest low Golgi function with moderate activation of caspase 3 pathway. Bottom row: exposure to 50 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). The majority of cells present intense, inhomogenuous, granular aspect of red fluorescence with lack of fluorescence staining suggesting an intense activation of caspase 3 apoptotic pathway. Magnification: 60×.Abbreviation: Alb-GNPs, albumin-conjugated gold nanoparticles.
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f5-ijn-10-5435: The concomitent detection of Golgi apparatus function and caspase 3 pathway.Notes: Upper row: control sample (no Alb-GNPs exposure: low red fluorescence is visible, suggesting that Hep G2 cells have low apoptotic state under normal conditions; in contrast Golgi function is normal as suggested by green fluorescence. Middle row: exposure to 10 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). Lower green fluorescence intensity staining combined with increased red fluorescence suggest low Golgi function with moderate activation of caspase 3 pathway. Bottom row: exposure to 50 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). The majority of cells present intense, inhomogenuous, granular aspect of red fluorescence with lack of fluorescence staining suggesting an intense activation of caspase 3 apoptotic pathway. Magnification: 60×.Abbreviation: Alb-GNPs, albumin-conjugated gold nanoparticles.

Mentions: As seen in Figure 5, we showed that PT ablation of Alb-GNPs causes a collapse of the GA. The ability of Alb-GNPs’ PT treatment to cause direct Golgi stress prompted us to next investigate the effects of this therapeutic method on caspase-3 pathway in malignant HepG2 cells. We first used fluorescence microscopy to display the cellular distribution of caspase-3 in HepG2 cells with or without Alb-GNPs treatment. In accordance with earlier reports in other malignant cells, the immunostaining pattern of caspase-3 was represented by punctuate structures homogenously localized in the cytoplasmic region. Following 1 hour of PT treatment with 50 μg/mL Alb-GNPs, caspase-3 lost its normal cellular localization pattern and appeared throughout the nucleus. The GA in these cells could no longer be visualized by the Golgi staining, strongly suggesting the collapse of this structure. Several reports have described a particular apoptotic pathway in malignant cells triggered by ER/Golgi stress and that further induces the mitochondrial cell death cascade.


Photothermal treatment of liver cancer with albumin-conjugated gold nanoparticles initiates Golgi Apparatus-ER dysfunction and caspase-3 apoptotic pathway activation by selective targeting of Gp60 receptor.

Mocan L, Matea C, Tabaran FA, Mosteanu O, Pop T, Mocan T, Iancu C - Int J Nanomedicine (2015)

The concomitent detection of Golgi apparatus function and caspase 3 pathway.Notes: Upper row: control sample (no Alb-GNPs exposure: low red fluorescence is visible, suggesting that Hep G2 cells have low apoptotic state under normal conditions; in contrast Golgi function is normal as suggested by green fluorescence. Middle row: exposure to 10 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). Lower green fluorescence intensity staining combined with increased red fluorescence suggest low Golgi function with moderate activation of caspase 3 pathway. Bottom row: exposure to 50 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). The majority of cells present intense, inhomogenuous, granular aspect of red fluorescence with lack of fluorescence staining suggesting an intense activation of caspase 3 apoptotic pathway. Magnification: 60×.Abbreviation: Alb-GNPs, albumin-conjugated gold nanoparticles.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4554431&req=5

f5-ijn-10-5435: The concomitent detection of Golgi apparatus function and caspase 3 pathway.Notes: Upper row: control sample (no Alb-GNPs exposure: low red fluorescence is visible, suggesting that Hep G2 cells have low apoptotic state under normal conditions; in contrast Golgi function is normal as suggested by green fluorescence. Middle row: exposure to 10 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). Lower green fluorescence intensity staining combined with increased red fluorescence suggest low Golgi function with moderate activation of caspase 3 pathway. Bottom row: exposure to 50 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). The majority of cells present intense, inhomogenuous, granular aspect of red fluorescence with lack of fluorescence staining suggesting an intense activation of caspase 3 apoptotic pathway. Magnification: 60×.Abbreviation: Alb-GNPs, albumin-conjugated gold nanoparticles.
Mentions: As seen in Figure 5, we showed that PT ablation of Alb-GNPs causes a collapse of the GA. The ability of Alb-GNPs’ PT treatment to cause direct Golgi stress prompted us to next investigate the effects of this therapeutic method on caspase-3 pathway in malignant HepG2 cells. We first used fluorescence microscopy to display the cellular distribution of caspase-3 in HepG2 cells with or without Alb-GNPs treatment. In accordance with earlier reports in other malignant cells, the immunostaining pattern of caspase-3 was represented by punctuate structures homogenously localized in the cytoplasmic region. Following 1 hour of PT treatment with 50 μg/mL Alb-GNPs, caspase-3 lost its normal cellular localization pattern and appeared throughout the nucleus. The GA in these cells could no longer be visualized by the Golgi staining, strongly suggesting the collapse of this structure. Several reports have described a particular apoptotic pathway in malignant cells triggered by ER/Golgi stress and that further induces the mitochondrial cell death cascade.

Bottom Line: Darkfield microscopy and immunochemical staining was used to demonstrate the selective internalization of Alb-GNPs inside the HepG2 cells via Gp60 receptors targeting.The postirradiation apoptotic rate of HepG2 cells treated with Alb-GNPs ranged from 25.8% (for 5 μg/mL) to 48.2% (for 50 μg/mL) at 60 seconds, while at 30 minutes the necrotic rate increased from 35.7% (5 μg/mL) to 52.3% (50 μg/mL), P-value <0.001.Significantly lower necrotic rates were obtained when human hepatocytes were treated with Alb-GNPs in a similar manner.

View Article: PubMed Central - PubMed

Affiliation: Nanomedicine Department, Regional Institute of Gastroenterology and Hepatology "Octavian Fodor", University of Medicine and Pharmacy, "Iuliu Hatieganu", Croitorilor, Cluj-Napoca, Romania ; Department of Surgery, University of Medicine and Pharmacy, "Iuliu Hatieganu", Croitorilor, Cluj-Napoca, Romania.

ABSTRACT
We present a method of enhanced laser thermal ablation of HepG2 cells based on a simple gold nanoparticle (GNP) carrier system such as serum albumin (Alb), and demonstrate its selective therapeutic efficacy compared with normal hepatocyte cells. HepG2 or hepatocytes were treated with Alb-GNPs at various concentrations and various incubation times, and further irradiated using a 2 W, 808 nm laser. Darkfield microscopy and immunochemical staining was used to demonstrate the selective internalization of Alb-GNPs inside the HepG2 cells via Gp60 receptors targeting. The postirradiation apoptotic rate of HepG2 cells treated with Alb-GNPs ranged from 25.8% (for 5 μg/mL) to 48.2% (for 50 μg/mL) at 60 seconds, while at 30 minutes the necrotic rate increased from 35.7% (5 μg/mL) to 52.3% (50 μg/mL), P-value <0.001. Significantly lower necrotic rates were obtained when human hepatocytes were treated with Alb-GNPs in a similar manner. We also showed by means of immunocytochemistry that photothermal treatment of Alb-conjugated GNPs in liver cancer initiates Golgi apparatus-endoplasmic reticulum dysfunction with consequent caspase-3 apoptotic pathway activation and cellular apoptosis. The presented results may become a new method of treating cancer cells by selective therapeutic vectors using nanolocalized thermal ablation by laser heating.

No MeSH data available.


Related in: MedlinePlus