Abstract
Introduction: Cancer is an uncontrollable proliferation of cells and poses a major public health
problem worldwide with the number of cases increasing each year. It results from alterations in
genetic and epigenetic factors that allow changes in cell proliferation, resistant to undergo
apoptosis, cell invasion and metastasis. The purpose of this study was to investigate the effect of
lso-mukaadial acetate (I MA) on human cancer in cell culture and animal model. The objective of
this study was to determine the therapeutic effect of lso-mukaadial acetate in breast cancer
xenografted mice model, pancreatic and colon cancer cell lines.
Materials and Methodology: In the in vivo study, female athymic nude mice were used and
inoculated with MCF-7 breast cancer cells subcutaneously. Group one served as a negative control
group (no treatments) and group two positive control group (cisplatin) which was administered
intravenously. IMA was administered orally to group three (100 mg/kg) and group four (300
mg/kg). Blood was collected (70 μL) from the tail vein on day zero, day one and day 3. Tumor
regression and body mass were measured every second day. Estimation of serum parameters for
renal Indices was examined using a creatinine assay. Histopathological analysis was conducted to
evaluate morphological changes of liver, kidney, spleen tissues and tumors before and after
compound administration under a light microscope. Apoptotic analysis using TUNEL system was
conducted on liver, kidney and spleen tissues.
In the in vitro study, MIA-PACA2 pancreatic cancer cells and HT29 colon cancer cells were
cultured and treated with IMA. Alamar blue reagent was used to conduct cell viability endpoint
assay for 24 hours and xCELLigence system was used to conduct real time cell viability for 60
hours after treatment with lso-mukaadial acetate. After obtaining ICsos, a cellular Adenosine
Triphosphate (ATP) assay was conducted after treatment with IMA using CellTiter-Glo ® 2.0
Reagent. Caspase Glo®-3/7 reagent was used to measure caspase 3/7 activity after treatment with
IMA. Mitochondrial dysfunction was tested using the Mitochondrial ToxGlo ™ reagent and
morphological changes were assessed by a light microscope after treatment with IMA.
Upregulation, and downregulation of selected genes in cancer cells were examined using RealTime
Polymerase Chain reaction (SYBR Green).
Results and Discussion: According to the in vivo findings, tumor shrinkage and reduction in body
mass were observed after treatment with IMA. Serum creatinine was slightly elevated after