التحري عن التشوهات الكروموسومية وطفرات المورث FLT3/ITD في مرضى ابيضاض الدم النخاعي الحاد == Screening For Chromosomal Aberrations And Gene Mutations FLT3/ITD In Patients With Acute Myeloid Leukemia
Author name:
سمارة كاظم محمد
Supervisor name:
عبد الحسين مويت الفيصل
General topic:
Biology
Specific topic:
Biotechnologies
Degree:
Master
University:
University of Baghdad - Institute Of Genetic Engineering And Biotechnology - Department Of Biotechnology
Language:
English
University location:
Baghdad
First pages:
24T2988 - p.pdf
Abstract:
صممت الدراسة الحالية من اجل تسليط الضوء على تاثيرات مرض ابيضاض الدم ((AMLفي بعض معايير الوراثة الخلوية والجزيئية الحاصلة في كريات الدم البيضاء في ثلاثة حالات للمرضى اشتملت على مرحلة ما قبل العلاج, في اثناء العلاج وبعد العلاج الكيميائي. كما شملت الدراسة ا | The present study was designed to shed light on the cytogenetic and molecular effects caused by acute myeloid leukemia (AML) pathogenesis in three stages before, during and after treatment with chemotherapy (in vitro) in lymphocytes. It was also aimed to investigate chromosomal aberrations, micronucleus formation and FLT3/ITD point mutations in CN - AML region 100 - 300 bp compared with healthy control. The study was comprised of forty - seven AML Iraqi patients their ages ranged between 2.5 - 81 years. It included twenty females and twenty seven males compared with twenty - six healthy control. The following results were obtained : AML is most common leukemia in adults and uncommon in children. It was found that 46.8% patients suffer from AML, who were under 15 years old, while 19.15% patients their age ranged between 16 - 30 years; 19.15% of patients their age were more than 45 years and 14.9% of patients their age ranged between 31 - 45 years of the total AML patients. AML is more common in males than females. The percentages of females and males were 42.5% and 57.5% respectively representing 1.35 : 1.00 male : female ratio. Two AML patients 5.3% have diploid cells when examined under light microscope. The highest mitotic index was 7.498±1.7 occurred in patients before taking treatment while recorded 6.784±4.5 during treatment and 7.000±2.5 after treatment. Nuclear anomalies such as nucleoplasmic bridges and nuclear buds were observed in AML patients, Micronucleus mean values recorded 0.033±0.018, 0.020±0.00015 and 0.036±0.01for AML patients before, during and after treatment respectively when compared with the control which recorded 0.002±0.00002. Nuclear division index (NDI) means before, during and after treatment of AML patients were 1.658±0.2, 1.000±0.022 and 1.424±0.19 respectively. Significant differences occurred among the three groups when compared with the control group (1.282±0.09). Extracted DNA from 30 AML patients was amplified by PCR to obtain FLT3/ITD gene from exon 11 to intron 12 and ten of them sent for sequencing. Two patients showed larger bands (470 and 460) bp when compared with wild type (330) bp. Among six patients, three displayed point mutations of deletion and substitution while others were normal since no mutations were detected. The percentages of mutation types were substitution 57.78% and deletion 22.2%. The effect of mutations was missense mutation (55.54%), deletion (22.22%) and nonsense (22.22%). It has been concluded from the current work that AML is more common in adult males, patients suffer from AML exhibited high MI, NDI, MN and point mutations are present in such patients including deletion and substitution causing missense, nonsense and deletion.
