Molecular Analysis of Induced Leukemia in Mice Treated with a Novel Bithiophene Derivative

Abstract

Leukemias are a group of hematological conditions characterized by the abnormal proliferation and development of WBCs. According to the degree of cell differentiation, they can be characterised as acute or chronic, and as myelocytic or lymphocytic, depending on the primary type of cell involved. There have been numerous genetic and environmental risk factors identified. Leukemia is the world's 15th most prevalent malignancy and the 11th main cause of cancer death. Males have a higher disease burden than females (incidence rate 6.1 vs. 4.3 per 100,000) and death rate (4.2 vs. 2.8 per 100,000 in males vs. females).The present study was designed to focus on the in vivo anticancer activity of a new bithiophene derivative, bithienyl fluorobenzamidine (BFB) against DMBA-induced leukemia in mice, as well as to determine the molecular mechanism by which it exerts its antiproliferative effects. To fulfill this aim, sixty male Swiss albino mice (45 days) were divided into six groups; normal control, leukemic control (DMBA group), BFB-treated 1/10, BFB-treated 1/5 and two groups of drug control. Mice were treated intravenously with 4 DMBA doses (30 mg/kg body weight) at intervals of 10 days, then divided into two groups to be treated with 17 intraperitoneal doses of BFB [1/10 LD50 and 1/5 LD50] through a month. Drug control groups were treated with 17 intraperitoneal administration of both BFB drug doses.The weights of mice were recorded weekly throughout the experimental period. At the end of the experiment, whole blood samples, bone marrow, liver and spleen were collected for analysis. The relative organ weights were calculated. Histological abnormalities were observed in liver and spleen by light microscope. Complete blood count (CBC) and bone marrow (BM) smear examination were assayed. The relative expression of bone marrow tumor protein 53 (p53) and cyclin dependent kinase 1 (CDK1), and cyclin dependent kinase inhibitor 1A (CDKN1A/p21) genes and their protein levels were quantified. The bone marrow Caspase-3 concentration was determined and protein levels of phoshorylated protein kinase B (p-AKT) and Phosphatase and tensin homologue deleted on chromosome 10 (PTEN) were measured.