Abilities of berberine and chemically modified berberines to interact with metformin and inhibit proliferation of pancreatic cancer cells

Abstract

A B S T R A C TPancreatic cancer is devastating cancer worldwide with few if any truly effective therapies. Pancreatic cancer has an increasing incidence and may become the second leading cause of death from cancer. Novel, more effective therapeutic approaches are needed as pancreatic cancer patients usually survive for less than a year after being diagnosed. Control of blood sugar levels by the prescription drug metformin in diseases such as diabetes mellitus has been examined in association with pancreatic cancer. While the clinical trials remain inconclusive, there is hope that certain diets and medications may affect positively the outcomes of patients with pancreatic and other cancers. Other natural compounds may share some of the effects of metformin. One "medicinal" fruit consumed by millions worldwide is berberine (BBR). Metformin and BBR both activate AMP-activated protein kinase (AMPK) which is a key mediator of glucose metabolism.

GSK-3β Can Regulate the Sensitivity of MIA-PaCa-2 Pancreatic and MCF-7 Breast Cancer Cells to Chemotherapeutic Drugs, Targeted Therapeutics and Nutraceuticals

“…The cells were recovered from a 65-year old Caucasian male PDAC patient [48]. MIA-PaCa-2 cells were cultured as described [49]. MCF-7 breast cancer cells (ATCC ® HTB-22 TM ) were obtained from the ATCC.…”

“…Cells were transfected the WT-GSK-3β and KD-GSK-3β DNAs as described [37] with Lipofectin (Invitrogen) as described by the manufacturer. The generation of MIA-PaCa-2 + pLXSN cells were previously described [49]. Further, 48 h after transfection, a selection medium (DMEM + 5% FBS or RPMI-1640 + 5% FBS + 2 mg/mL G418 (Geneticin) (Invitrogen) was added to isolate stably transfected cells.…”

“…2.4. Cell Proliferation Assays in the Presence of Chemotherapeutic Drugs, Signal Transduction Inhibitors, and Nutraceuticals MIA-PaCa-2 + WT-GSK-3β, MIA-PaCa-2 + KD-GSK-3β, MIA-PaCa-2 + pLXSN, MCF-7 + WT-GSK-3β, MCF-7 + KD-GSK-3β, and MCF-7 + pLXSN cells were seeded in 96-well cell culture plates (BD Biosciences, San Jose, CA) at a density of 5000 cells/well in 100 µL of phenol red free RPMI-1640 containing 1% FBS as described [37,49]. The treatment medium was prepared by performing 10 two-fold serial dilutions to create a range of 11 concentrations of the different drugs, signal transduction inhibitors, and nutraceuticals.…”

“…The treatment medium was prepared by performing 10 two-fold serial dilutions to create a range of 11 concentrations of the different drugs, signal transduction inhibitors, and nutraceuticals. After 72 h of treatment (four days after seeding), the amount of 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) (Sigma-Aldrich) reduction in each well was quantified as described [37,49]. The absorbance at 570 nM was determined with a FL600 microplate fluorescence reader (Bio-Tek Instruments; Winooski, VT, USA) as described [37,49].…”

“…After 72 h of treatment (four days after seeding), the amount of 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) (Sigma-Aldrich) reduction in each well was quantified as described [37,49]. The absorbance at 570 nM was determined with a FL600 microplate fluorescence reader (Bio-Tek Instruments; Winooski, VT, USA) as described [37,49]. The mean and corresponding standard deviation of normalized adjusted absorbance was calculated from three replicate wells for each drug concentration.…”