Human Pancreatic Cell Line BxPC-3 Research Articles

Preparation of albumin nanospheres loaded with gemcitabine and their cytotoxicity against BXPC-3 cells in vitro

To optimize formulation methods for loading gemcitabine (GEM), the main drug against pancreatic cancer, into albumin nanoparticles for extended blood circulation and improved efficacy. GEM was loaded into two sizes of disolvation-crosslinked bovine serum albumin nanoparticles, with a mean diameter of 109.7 nm and 405.6 nm, respectively, by co-precipitation (the direct method) and follow-up adsorption (the indirect method). The antitumor activities of the two nanoparticulate formulations, were evaluated according to their anti-proliferative effects on the human pancreatic cell line BXPC-3, which were assessed using the MTT assay. The two nanoparticulate formulations, created by direct co-precipitation and indirect adsorption, possessed smooth surfaces and high drug loading efficiencies, 83% and 93% at 11% and 13% drug loading, respectively. The two formulations released GEM for 8 and 12 h, respectively, and significantly improved anti-BXPC-3 proliferation effects, as compared with the GEM solution and the drug-free albumin particles. Co-precipitating and adsorbing GEM into albumin particles resulted in sustained-release nanoparticulate formulations with improved antitumor cytotoxicity. The result suggests that this is a useful formulation strategy for improving the antitumor efficacy of GEM.

CURCUMIN SENSITIZES PANCREATIC CANCER CELLS TO APOG2 INDUCED KILLING

Objectives: In Pancreatic cancer, several important survival molecules such as NF-κB, a key transcription factor playing important roles in tumorigenesis, metastasis and inflammation is highly upregulated along with NF-κB downstream proteins like Bcl-2 and Bcl-xL. Thus novel agents that have the capacity of inactivating such crucial target proteins could enhance the antitumor activity. ApoG2, new derivative of natural product gossypol, is a potent small molecule inhibitor of Bcl-2 and Bcl-xL proteins. Curcumin, an active ingredient from the rhizome of the plant, Curcuma longa, has anti oxidant, anti-inflammatory and anti-cancer activities and works by inactivation of NF-κB activity. In this study we have tested our hypothesis whether inactivation of NF-κB by curcumin could enhance anti-tumor activity of ApoG2. Methods: The BxPC-3 human pancreatic cell line was used in this study. For cell viability, apoptosis and NF-κB studies, MTT assay, ELISA and electrophoretic mobility shift assay were used respectively. Western blot assays was done to study Bcl-2 and Bcl-xL. Results: Treatment of human pancreatic cell line BxPC-3 with curcumin sensitized ApoG2 induced cell killing. In the dose escalation study with ApoG2 at 72 hrs, 1μM showed 60% cell viability, while 5μM of curcumin at 72 hrs showed 78 % cell viability. Combination treatment with 1μM of ApoG2 and 5μM of curcumin at 72 hrs showed 45 % cell viability. (P = <0.0001). Apoptosis by Histone/DNA ELISA showed 7-fold induction with ApoG2 and 6-fold induction with curcumin at 72 hrs. However, combination treatment with curcumin and ApoG2 at 72 hrs showed 11-fold induction in apoptosis compared to the control. Bcl-xL and Bcl-2 expression levels were down regulated along with the inactivation of NF-κB in the combination treatment group compared with curcumin and ApoG2 alone. Conclusion: From these results, we conclude that curcumin enhanced ApoG2 induced cell growth inhibition and apoptosis of Pancreatic BxPC-3 cancer cells. This effect is partly due to inactivation of NF-κB activation by curcumin and inhibition of Bcl-2 and Bcl-xL.

Sensitization of pancreatic cancer cells to ApoG2 induced killing by curcumin

13152 Background: In pancreatic cancer, several important survival molecules such as NF-kappa B and Bcl-2 or Bcl-XL are highly activated. Thus agents that inhibit NF-KappaB activation, together with agents that inhibit Bcl-2 or Bcl-XL protein function, may lead to enhanced cell killing. ApoG2, a new derivative of natural product gossypol, is a potent small molecule inhibitor of Bcl-2 and Bcl-XL proteins. Curcumin, an active ingredient from the rhizome of the plant, Curcuma longa, has anti-inflammatory and anti-cancer activities and works by inactivation of NF-Kappa B activity. In this study we have investigated the effects of curcumin on ApoG2 induced apoptosis. Methods: The BxPC-3 human pancreatic cancer cell line was used in this study. ApoG2 was dissolved in DMSO at 1 mM stock solution, and curcumin was dissolved in DMSO at 20 mM stock solution. Cell viability was determined by the standard MTT assay and apoptosis was measured by histone/DNA ELISA method. Results: Pretreatment of human pancreatic cell line BxPC-3 with curcumin sensitizes ApoG2 induced cell killing. In the dose escalation with ApoG2 at 48 hrs on MTT assay, 250 nM showed 75% cell viability, while 2.5 uM of curcumin at 72 hrs showed 80% cell viability. Pretreatment with curcumin for 24 hrs followed by ApoG2 at 48 hrs showed 55% cell viability (p = 0.0005). Apoptosis by histone/DNA ELISA showed 4-fold induction of cell death with ApoG2 at 48 hrs, 7-fold induction of cell death with curcumin at 72 hrs. However, pretreatment with curcumin for 24 hrs followed by ApoG2 at 48 hrs showed 10-fold induction in apoptosis compared to the control. Conclusions: From these results, we conclude that curcumin enhanced ApoG2 induced cell growth inhibition and apoptosis of BxPC-3 cells. No significant financial relationships to disclose.

Characterization of the oligosaccharide moiety of VIP receptor from the human pancreatic cell line BxPC-3

The human pancreatic cell line BxPC-3 displays two classes of binding sites with high and low affinity for VIP. The order of potency of VIP-related peptides in inhibiting either [ 125I]VIP or [ 125I] N- AcPACAP27 binding and in stimulating cAMP production was typical of the human VIP receptor. By combining affinity labeling with glycosidase treatments, we have characterized the VIP receptor as a M r = 68,200 glycoprotein, consisting of a M r = 39,300 polypeptide core with at least three N- linked oligosaccharide chains. In addition, our results revealed the presence of a low amount of sialic acid residues in the carbohydrate moiety of receptor.