Limits...
Characterization of 9-nitrocamptothecin liposomes: anticancer properties and mechanisms on hepatocellular carcinoma in vitro and in vivo.

Zheng S, Chang S, Lu J, Chen Z, Xie L, Nie Y, He B, Zou S, Gu Z - PLoS ONE (2011)

Bottom Line: Then we compared the effects of 9NC and 9NC-LP on the proliferation and apoptosis of HepG2, Bel-7402, Hep3B and L02 cells in vitro.Exposure to 9NC-LP led to increased expression of p53, p21, p27, Bax, caspase-3, caspase-8, caspase-9 and apoptosis-inducing factor, mitochondrion-associated 1 and decreased expression of Bcl-2, cyclin E, cyclin A, Cdk2 and cyclin D1.In conclusion, 9NC and 9NC-LP can inhibit HCC growth via cell cycle arrest and induction of apoptosis. 9NC-LP has a more potent anti-tumor effect and fewer side effects in vivo, which means it is a promising reagent for cancer therapy via intravenous administration.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan, People's Republic of China.

ABSTRACT

Background: Hepatocellular carcinoma (HCC) is the third most common cause of cancer related mortality worldwide. 9-Nitrocamptothecin (9NC) is a potent topoisomerase-I inhibitor with strong anticancer effect. To increase the solubility and stability, we synthesized a novel 9NC loaded liposomes (9NC-LP) via incorporating 9NC into liposomes. In the present study, we determined the effects of 9NC and 9NC-LP on in vitro and in vivo, and the underlying mechanisms.

Methodology/principal findings: We first analyzed the characteristics of 9NC-LP. Then we compared the effects of 9NC and 9NC-LP on the proliferation and apoptosis of HepG2, Bel-7402, Hep3B and L02 cells in vitro. We also investigated their anticancer properties in nude mice bearing HCC xenograft in vivo. 9NC-LP has a uniform size (around 190 nm) and zeta potential (∼-11 mV), and exhibited a steady sustained-release pattern profile in vitro. Both 9NC and 9NC-LP could cause cell cycle arrest and apoptosis in a dose-dependent and p53-dependent manner. However, this effect was not ubiquitous in all cell lines. Exposure to 9NC-LP led to increased expression of p53, p21, p27, Bax, caspase-3, caspase-8, caspase-9 and apoptosis-inducing factor, mitochondrion-associated 1 and decreased expression of Bcl-2, cyclin E, cyclin A, Cdk2 and cyclin D1. Furthermore, 9NC-LP exhibited a more potent antiproliferative effect and less side effects in vivo. Western blot analysis of the xenograft tumors in nude mice showed similar changes in protein expression in vivo.

Conclusions/significance: In conclusion, 9NC and 9NC-LP can inhibit HCC growth via cell cycle arrest and induction of apoptosis. 9NC-LP has a more potent anti-tumor effect and fewer side effects in vivo, which means it is a promising reagent for cancer therapy via intravenous administration.

Show MeSH

Related in: MedlinePlus

Cell cycle arrest and apoptosis-induced effects of 9NC and 9NC-LP on HepG2, L02 and Hep3B cells in vitro.For the cell cycle analysis, cells were seeded at 2×105 to 3×105 per well in six-well paltes in triplicates and incubated with compounds. After treated for 24 h, 48 h and 72 h, cells were fixed, permeabilized, stained with PI and analyzed by flow cytometry. Results showed cells were incubated with 0, 0.01, 0.1, 1 µmol/L 9NC-LP for 24 h(A). Apoptosis of HepG2, L02 and Hep3B cells(B) was measured by flow cytometry after stained with AnnexinV-PE and 7-AAD (Concentration: 0, 0.1, 1, 10 µmol/L). This experiment was done in triplicate and representative diagrams are shown (There no significant differences between cells treated with 4‰DMSO, Liposomes (free) and untreated cells. Data not shown).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3111480&req=5

pone-0021064-g003: Cell cycle arrest and apoptosis-induced effects of 9NC and 9NC-LP on HepG2, L02 and Hep3B cells in vitro.For the cell cycle analysis, cells were seeded at 2×105 to 3×105 per well in six-well paltes in triplicates and incubated with compounds. After treated for 24 h, 48 h and 72 h, cells were fixed, permeabilized, stained with PI and analyzed by flow cytometry. Results showed cells were incubated with 0, 0.01, 0.1, 1 µmol/L 9NC-LP for 24 h(A). Apoptosis of HepG2, L02 and Hep3B cells(B) was measured by flow cytometry after stained with AnnexinV-PE and 7-AAD (Concentration: 0, 0.1, 1, 10 µmol/L). This experiment was done in triplicate and representative diagrams are shown (There no significant differences between cells treated with 4‰DMSO, Liposomes (free) and untreated cells. Data not shown).

Mentions: 9NC and 9NC-LP could induce apoptosis (increased Sub G1 peak) (p<0.05) and cause cell cycle arrest (p<0.05) of the HCC cell lines. The effect is dose- and time-dependent in all tested cell lines. Cell cycle phase is alternation according to the compound concentration and incubation time in all test cell lines. S phase delay was observed after exposure for 24 h and G2/M phase delay was observed after exposure for 72 h in all tested cell lines (Fig. 3A, Table S1). Both compounds caused more significant cell cycle arrest in the HepG2 cells than in other cell lines. Almost all of the HepG2 and Bel-7402 cells arrest in S phase when compounds concentrations of both compounds were over 0.05 µmol/L while L02 and Hep3B cells were more resistant.


Characterization of 9-nitrocamptothecin liposomes: anticancer properties and mechanisms on hepatocellular carcinoma in vitro and in vivo.

Zheng S, Chang S, Lu J, Chen Z, Xie L, Nie Y, He B, Zou S, Gu Z - PLoS ONE (2011)

Cell cycle arrest and apoptosis-induced effects of 9NC and 9NC-LP on HepG2, L02 and Hep3B cells in vitro.For the cell cycle analysis, cells were seeded at 2×105 to 3×105 per well in six-well paltes in triplicates and incubated with compounds. After treated for 24 h, 48 h and 72 h, cells were fixed, permeabilized, stained with PI and analyzed by flow cytometry. Results showed cells were incubated with 0, 0.01, 0.1, 1 µmol/L 9NC-LP for 24 h(A). Apoptosis of HepG2, L02 and Hep3B cells(B) was measured by flow cytometry after stained with AnnexinV-PE and 7-AAD (Concentration: 0, 0.1, 1, 10 µmol/L). This experiment was done in triplicate and representative diagrams are shown (There no significant differences between cells treated with 4‰DMSO, Liposomes (free) and untreated cells. Data not shown).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3111480&req=5

pone-0021064-g003: Cell cycle arrest and apoptosis-induced effects of 9NC and 9NC-LP on HepG2, L02 and Hep3B cells in vitro.For the cell cycle analysis, cells were seeded at 2×105 to 3×105 per well in six-well paltes in triplicates and incubated with compounds. After treated for 24 h, 48 h and 72 h, cells were fixed, permeabilized, stained with PI and analyzed by flow cytometry. Results showed cells were incubated with 0, 0.01, 0.1, 1 µmol/L 9NC-LP for 24 h(A). Apoptosis of HepG2, L02 and Hep3B cells(B) was measured by flow cytometry after stained with AnnexinV-PE and 7-AAD (Concentration: 0, 0.1, 1, 10 µmol/L). This experiment was done in triplicate and representative diagrams are shown (There no significant differences between cells treated with 4‰DMSO, Liposomes (free) and untreated cells. Data not shown).
Mentions: 9NC and 9NC-LP could induce apoptosis (increased Sub G1 peak) (p<0.05) and cause cell cycle arrest (p<0.05) of the HCC cell lines. The effect is dose- and time-dependent in all tested cell lines. Cell cycle phase is alternation according to the compound concentration and incubation time in all test cell lines. S phase delay was observed after exposure for 24 h and G2/M phase delay was observed after exposure for 72 h in all tested cell lines (Fig. 3A, Table S1). Both compounds caused more significant cell cycle arrest in the HepG2 cells than in other cell lines. Almost all of the HepG2 and Bel-7402 cells arrest in S phase when compounds concentrations of both compounds were over 0.05 µmol/L while L02 and Hep3B cells were more resistant.

Bottom Line: Then we compared the effects of 9NC and 9NC-LP on the proliferation and apoptosis of HepG2, Bel-7402, Hep3B and L02 cells in vitro.Exposure to 9NC-LP led to increased expression of p53, p21, p27, Bax, caspase-3, caspase-8, caspase-9 and apoptosis-inducing factor, mitochondrion-associated 1 and decreased expression of Bcl-2, cyclin E, cyclin A, Cdk2 and cyclin D1.In conclusion, 9NC and 9NC-LP can inhibit HCC growth via cell cycle arrest and induction of apoptosis. 9NC-LP has a more potent anti-tumor effect and fewer side effects in vivo, which means it is a promising reagent for cancer therapy via intravenous administration.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan, People's Republic of China.

ABSTRACT

Background: Hepatocellular carcinoma (HCC) is the third most common cause of cancer related mortality worldwide. 9-Nitrocamptothecin (9NC) is a potent topoisomerase-I inhibitor with strong anticancer effect. To increase the solubility and stability, we synthesized a novel 9NC loaded liposomes (9NC-LP) via incorporating 9NC into liposomes. In the present study, we determined the effects of 9NC and 9NC-LP on in vitro and in vivo, and the underlying mechanisms.

Methodology/principal findings: We first analyzed the characteristics of 9NC-LP. Then we compared the effects of 9NC and 9NC-LP on the proliferation and apoptosis of HepG2, Bel-7402, Hep3B and L02 cells in vitro. We also investigated their anticancer properties in nude mice bearing HCC xenograft in vivo. 9NC-LP has a uniform size (around 190 nm) and zeta potential (∼-11 mV), and exhibited a steady sustained-release pattern profile in vitro. Both 9NC and 9NC-LP could cause cell cycle arrest and apoptosis in a dose-dependent and p53-dependent manner. However, this effect was not ubiquitous in all cell lines. Exposure to 9NC-LP led to increased expression of p53, p21, p27, Bax, caspase-3, caspase-8, caspase-9 and apoptosis-inducing factor, mitochondrion-associated 1 and decreased expression of Bcl-2, cyclin E, cyclin A, Cdk2 and cyclin D1. Furthermore, 9NC-LP exhibited a more potent antiproliferative effect and less side effects in vivo. Western blot analysis of the xenograft tumors in nude mice showed similar changes in protein expression in vivo.

Conclusions/significance: In conclusion, 9NC and 9NC-LP can inhibit HCC growth via cell cycle arrest and induction of apoptosis. 9NC-LP has a more potent anti-tumor effect and fewer side effects in vivo, which means it is a promising reagent for cancer therapy via intravenous administration.

Show MeSH
Related in: MedlinePlus