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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 and ER function following Alb-GNPs mediated photothermal treatment.Notes: Exposure to 10 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). Upper row: HepB5 cells: normal red and green fluorescence is visible, suggesting that HepB5 cells function normally. Third row: HepG2 cells: red and green fluorescence is less visible, with mild architectural changes in both ER and Golgi. Second row: exposure to 50 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). Bottom row: HepB5 cells: normal green fluorescence suggest a normal function of the GA, with mild disturbance of ER function. HepG2 cells: red and green fluorescence is less visible, with severe architectural changes in both ER and GA suggesting lack of function in these organelles. Magnification: 60×.Abbreviations: ER, endoplasmic reticulum; GA, Golgi apparatus; Alb-GNPs, albumin-conjugated gold nanoparticles.
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f3-ijn-10-5435: The concomitent detection of Golgi apparatus and ER function following Alb-GNPs mediated photothermal treatment.Notes: Exposure to 10 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). Upper row: HepB5 cells: normal red and green fluorescence is visible, suggesting that HepB5 cells function normally. Third row: HepG2 cells: red and green fluorescence is less visible, with mild architectural changes in both ER and Golgi. Second row: exposure to 50 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). Bottom row: HepB5 cells: normal green fluorescence suggest a normal function of the GA, with mild disturbance of ER function. HepG2 cells: red and green fluorescence is less visible, with severe architectural changes in both ER and GA suggesting lack of function in these organelles. Magnification: 60×.Abbreviations: ER, endoplasmic reticulum; GA, Golgi apparatus; Alb-GNPs, albumin-conjugated gold nanoparticles.

Mentions: As seen in Figure 3, we observed that photothermal (PT) treatment mediated by Alb-GNPs causes a collapse of the GA. The disruptive effect on the GA ultimately leads to malfunction of ER in HepG2 cells. To analyze this hypothesis, cells were treated at various concentrations and various incubation times, and further stained specifically for analysis of changes in ER morphology by fluorescence microscopy. As shown in Figure 3, PT treatment caused a severe, concentration-dependent dilation of ER membranes. ImageJ quantification of fluorescent areas revealed a significant decrease (P<0.005) in both red and green fluorescence following Alb-GNPs mediated PT treatment in HepG2 cells, but not in control HepB5 cells. This dramatic change in ER morphology occurred prior to the onset of appreciable apoptosis and did not occur in HepB5-treated cells, strongly suggesting that these effects were specific to cancer liver treatment and not a general feature of any type of cell. Taken together, these data suggest that HepG2 cells may be intrinsically sensitive to this particular type of GNP-mediated thermal ablation.


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 and ER function following Alb-GNPs mediated photothermal treatment.Notes: Exposure to 10 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). Upper row: HepB5 cells: normal red and green fluorescence is visible, suggesting that HepB5 cells function normally. Third row: HepG2 cells: red and green fluorescence is less visible, with mild architectural changes in both ER and Golgi. Second row: exposure to 50 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). Bottom row: HepB5 cells: normal green fluorescence suggest a normal function of the GA, with mild disturbance of ER function. HepG2 cells: red and green fluorescence is less visible, with severe architectural changes in both ER and GA suggesting lack of function in these organelles. Magnification: 60×.Abbreviations: ER, endoplasmic reticulum; GA, Golgi apparatus; Alb-GNPs, albumin-conjugated gold nanoparticles.
© Copyright Policy
Related In: Results  -  Collection

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

f3-ijn-10-5435: The concomitent detection of Golgi apparatus and ER function following Alb-GNPs mediated photothermal treatment.Notes: Exposure to 10 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). Upper row: HepB5 cells: normal red and green fluorescence is visible, suggesting that HepB5 cells function normally. Third row: HepG2 cells: red and green fluorescence is less visible, with mild architectural changes in both ER and Golgi. Second row: exposure to 50 μg/mL Alb-GNPs (1 hour, 37°C), followed by laser excitation (3 minutes, 808 nm, 2W/cm2). Bottom row: HepB5 cells: normal green fluorescence suggest a normal function of the GA, with mild disturbance of ER function. HepG2 cells: red and green fluorescence is less visible, with severe architectural changes in both ER and GA suggesting lack of function in these organelles. Magnification: 60×.Abbreviations: ER, endoplasmic reticulum; GA, Golgi apparatus; Alb-GNPs, albumin-conjugated gold nanoparticles.
Mentions: As seen in Figure 3, we observed that photothermal (PT) treatment mediated by Alb-GNPs causes a collapse of the GA. The disruptive effect on the GA ultimately leads to malfunction of ER in HepG2 cells. To analyze this hypothesis, cells were treated at various concentrations and various incubation times, and further stained specifically for analysis of changes in ER morphology by fluorescence microscopy. As shown in Figure 3, PT treatment caused a severe, concentration-dependent dilation of ER membranes. ImageJ quantification of fluorescent areas revealed a significant decrease (P<0.005) in both red and green fluorescence following Alb-GNPs mediated PT treatment in HepG2 cells, but not in control HepB5 cells. This dramatic change in ER morphology occurred prior to the onset of appreciable apoptosis and did not occur in HepB5-treated cells, strongly suggesting that these effects were specific to cancer liver treatment and not a general feature of any type of cell. Taken together, these data suggest that HepG2 cells may be intrinsically sensitive to this particular type of GNP-mediated thermal ablation.

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