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A liposomal formulation of the synthetic curcumin analog EF24 (Lipo-EF24) inhibits pancreatic cancer progression: towards future combination therapies.

Bisht S, Schlesinger M, Rupp A, Schubert R, Nolting J, Wenzel J, Holdenrieder S, Brossart P, Bendas G, Feldmann G - J Nanobiotechnology (2016)

Bottom Line: Lipo-EF24 potently suppressed NF-kappaB nuclear translocation by inhibiting phosphorylation and subsequent degradation of its inhibitor I-kappa-B-alpha.In vivo, synergistic tumor growth inhibition was observed in MIAPaCa xenografts when Lipo-EF24 was given in combination with the standard-of-care cytotoxic agent gemcitabine.In line with in vitro observations, western blot analysis revealed decreased phosphorylation of I-kappa-B-alpha in excised Lipo-EF24-treated xenograft tumor tissues.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine 3, Center of Integrated Oncology (CIO) Cologne-Bonn, University Hospital of Bonn, Sigmund-Freud-Str. 25, 53127, Bonn, Germany.

ABSTRACT

Background: Pancreatic cancer is one of the most lethal of human malignancies known to date and shows relative insensitivity towards most of the clinically available therapy regimens. 3,5-bis(2-fluorobenzylidene)-4-piperidone (EF24), a novel synthetic curcumin analog, has shown promising in vitro therapeutic efficacy in various human cancer cells, but insufficient water solubility and systemic bioavailability limit its clinical application. Here, we describe nano-encapsulation of EF24 into pegylated liposomes (Lipo-EF24) and evaluation of these particles in preclinical in vitro and in vivo model systems of pancreatic cancer.

Results: Transmission electron microscopy and size distribution studies by dynamic light scattering confirmed intact spherical morphology of the formed liposomes with an average diameter of less than 150 nm. In vitro, treatment with Lipo-EF24 induced growth inhibition and apoptosis in MIAPaCa and Pa03C pancreatic cancer cells as assessed by using cell viability and proliferation assays, replating and soft agar clonogenicity assays as well as western blot analyses. Lipo-EF24 potently suppressed NF-kappaB nuclear translocation by inhibiting phosphorylation and subsequent degradation of its inhibitor I-kappa-B-alpha. In vivo, synergistic tumor growth inhibition was observed in MIAPaCa xenografts when Lipo-EF24 was given in combination with the standard-of-care cytotoxic agent gemcitabine. In line with in vitro observations, western blot analysis revealed decreased phosphorylation of I-kappa-B-alpha in excised Lipo-EF24-treated xenograft tumor tissues.

Conclusion: Due to its promising therapeutic efficacy and favorable toxicity profile Lipo-EF24 might be a promising starting point for development of future combinatorial therapeutic regimens against pancreatic cancer.

No MeSH data available.


Related in: MedlinePlus

Lipo-EF24 inhibits growth and clonogenicity of pancreatic cancer cell lines. a MTS assays were performed using equivalent doses of free and liposomal EF24 in two different pancreatic cancer cell lines, MIAPaCa and Pa03C. In both cell lines, liposomal EF24 demonstrated marked growth suppression comparable to that of free drug. Further, liposomal EF24 caused significant, dose-dependent reduction in the ability of these cells to form colonies in replating assays (b) and inhibited anchorage-independent growth in soft agar (c). Void liposomes showed no significant effect in any of these cell lines. Representative images from one of three independent experiments are shown
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Fig3: Lipo-EF24 inhibits growth and clonogenicity of pancreatic cancer cell lines. a MTS assays were performed using equivalent doses of free and liposomal EF24 in two different pancreatic cancer cell lines, MIAPaCa and Pa03C. In both cell lines, liposomal EF24 demonstrated marked growth suppression comparable to that of free drug. Further, liposomal EF24 caused significant, dose-dependent reduction in the ability of these cells to form colonies in replating assays (b) and inhibited anchorage-independent growth in soft agar (c). Void liposomes showed no significant effect in any of these cell lines. Representative images from one of three independent experiments are shown

Mentions: The therapeutic efficacy of liposomal EF24 was examined in two different human pancreatic cancer lines (MIAPaCa and Pa03C) and directly compared to free EF24 or void liposomes, respectively, using MTS assay. As shown in Fig. 3a, Lipo-EF24 significantly inhibited growth of both cell lines in a dose-dependent manner and to an extent that was comparable to that of free EF24 while void liposomes showed negligible effects on cell growth. Moreover, when assessed using replating assays, both the cell lines mentioned above failed to grow colonies from single cell suspensions in the presence of liposomal EF24 but readily formed colonies when exposed to void liposomes (Fig. 3b). Likewise, liposomal EF24 potently and reproducibly abrogated anchorage independent growth of MIAPaCa and Pa03C cells in softagar, while such an effect was not observed for void liposomes or untreated controls (Fig. 3c). Next, the effect of liposomal EF24 on cellular proliferation was analyzed using a CFSE dilution assay, which relies on the depletion of fluorescence intensity of CFSE with cell division. Of note, addition of Lipo-EF24 to culture media at concentrations of 5 or 10 µM on CFSE labeled cells for 48 h significantly decreased proliferation of both cell lines as compared to controls (Fig. 4a).Fig. 3


A liposomal formulation of the synthetic curcumin analog EF24 (Lipo-EF24) inhibits pancreatic cancer progression: towards future combination therapies.

Bisht S, Schlesinger M, Rupp A, Schubert R, Nolting J, Wenzel J, Holdenrieder S, Brossart P, Bendas G, Feldmann G - J Nanobiotechnology (2016)

Lipo-EF24 inhibits growth and clonogenicity of pancreatic cancer cell lines. a MTS assays were performed using equivalent doses of free and liposomal EF24 in two different pancreatic cancer cell lines, MIAPaCa and Pa03C. In both cell lines, liposomal EF24 demonstrated marked growth suppression comparable to that of free drug. Further, liposomal EF24 caused significant, dose-dependent reduction in the ability of these cells to form colonies in replating assays (b) and inhibited anchorage-independent growth in soft agar (c). Void liposomes showed no significant effect in any of these cell lines. Representative images from one of three independent experiments are shown
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4940769&req=5

Fig3: Lipo-EF24 inhibits growth and clonogenicity of pancreatic cancer cell lines. a MTS assays were performed using equivalent doses of free and liposomal EF24 in two different pancreatic cancer cell lines, MIAPaCa and Pa03C. In both cell lines, liposomal EF24 demonstrated marked growth suppression comparable to that of free drug. Further, liposomal EF24 caused significant, dose-dependent reduction in the ability of these cells to form colonies in replating assays (b) and inhibited anchorage-independent growth in soft agar (c). Void liposomes showed no significant effect in any of these cell lines. Representative images from one of three independent experiments are shown
Mentions: The therapeutic efficacy of liposomal EF24 was examined in two different human pancreatic cancer lines (MIAPaCa and Pa03C) and directly compared to free EF24 or void liposomes, respectively, using MTS assay. As shown in Fig. 3a, Lipo-EF24 significantly inhibited growth of both cell lines in a dose-dependent manner and to an extent that was comparable to that of free EF24 while void liposomes showed negligible effects on cell growth. Moreover, when assessed using replating assays, both the cell lines mentioned above failed to grow colonies from single cell suspensions in the presence of liposomal EF24 but readily formed colonies when exposed to void liposomes (Fig. 3b). Likewise, liposomal EF24 potently and reproducibly abrogated anchorage independent growth of MIAPaCa and Pa03C cells in softagar, while such an effect was not observed for void liposomes or untreated controls (Fig. 3c). Next, the effect of liposomal EF24 on cellular proliferation was analyzed using a CFSE dilution assay, which relies on the depletion of fluorescence intensity of CFSE with cell division. Of note, addition of Lipo-EF24 to culture media at concentrations of 5 or 10 µM on CFSE labeled cells for 48 h significantly decreased proliferation of both cell lines as compared to controls (Fig. 4a).Fig. 3

Bottom Line: Lipo-EF24 potently suppressed NF-kappaB nuclear translocation by inhibiting phosphorylation and subsequent degradation of its inhibitor I-kappa-B-alpha.In vivo, synergistic tumor growth inhibition was observed in MIAPaCa xenografts when Lipo-EF24 was given in combination with the standard-of-care cytotoxic agent gemcitabine.In line with in vitro observations, western blot analysis revealed decreased phosphorylation of I-kappa-B-alpha in excised Lipo-EF24-treated xenograft tumor tissues.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine 3, Center of Integrated Oncology (CIO) Cologne-Bonn, University Hospital of Bonn, Sigmund-Freud-Str. 25, 53127, Bonn, Germany.

ABSTRACT

Background: Pancreatic cancer is one of the most lethal of human malignancies known to date and shows relative insensitivity towards most of the clinically available therapy regimens. 3,5-bis(2-fluorobenzylidene)-4-piperidone (EF24), a novel synthetic curcumin analog, has shown promising in vitro therapeutic efficacy in various human cancer cells, but insufficient water solubility and systemic bioavailability limit its clinical application. Here, we describe nano-encapsulation of EF24 into pegylated liposomes (Lipo-EF24) and evaluation of these particles in preclinical in vitro and in vivo model systems of pancreatic cancer.

Results: Transmission electron microscopy and size distribution studies by dynamic light scattering confirmed intact spherical morphology of the formed liposomes with an average diameter of less than 150 nm. In vitro, treatment with Lipo-EF24 induced growth inhibition and apoptosis in MIAPaCa and Pa03C pancreatic cancer cells as assessed by using cell viability and proliferation assays, replating and soft agar clonogenicity assays as well as western blot analyses. Lipo-EF24 potently suppressed NF-kappaB nuclear translocation by inhibiting phosphorylation and subsequent degradation of its inhibitor I-kappa-B-alpha. In vivo, synergistic tumor growth inhibition was observed in MIAPaCa xenografts when Lipo-EF24 was given in combination with the standard-of-care cytotoxic agent gemcitabine. In line with in vitro observations, western blot analysis revealed decreased phosphorylation of I-kappa-B-alpha in excised Lipo-EF24-treated xenograft tumor tissues.

Conclusion: Due to its promising therapeutic efficacy and favorable toxicity profile Lipo-EF24 might be a promising starting point for development of future combinatorial therapeutic regimens against pancreatic cancer.

No MeSH data available.


Related in: MedlinePlus