Molecular Evaluating the Therapeutic Application of Anticancer L-glutaminase Enzyme on Genetic Mutation Induced Rats

Abstract

The Genomic DNA had been extracted from the liver tissue of the experimental rat groups following 15 weeks of treatment with partial purified L-glutaminase enzyme from E.coli in relation with ethylamine. The DNA samples of all the six treatment groups were amplified by PCR using three different primers specific for determining the presence of P53, bax and G3pdh genes and detect the effect of Ethylenimine and L-glutaminase on the animals at the molecular level. The agarose gel electrophoresis technique used to analyze the PCR amplification products. The result revealed the presence of P53 gene in all treatment groups except for diseased group (T2). This finding demonstrates the mutagenic effect of Ethylenimine that lead to mutation at P53 gene sequence, and therapeutic beneficial of L-glutaminase. Also, there is no PCR amplification product which represent Bax gene sequence for T2 and T5 groups which were administered doses of Ethylenimine, indicating that low doses of L-glutaminase failed to prevent the mutagenic effect of Ethylenimine. While, the G3pd genes is presented only in T2 and T5. Finally, analysis the DNA sequences of the PCR amplified products extracted from liver samples of T2 group treated with Ethylenimine was carried out, then the results were compared with NCBI. The expected mutations were found at thirteen locals and there were only three mutated sequences with the DNA of liver samples from T5 group, while the DNA of animals in group (T6) were showed no mutated regions. The results of this novel study make clear the therapeutic effect of L-glutaminase and how suppress the mutagenic and carcinogenic effect of ethylenimine on P53, Bax, and G3pd genes, and its effect was dose dependent..