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Screening of C-kit gene Mutation in Acute Myeloid Leukaemia in Northern India.

Hussain SR, Raza ST, Babu SG, Singh P, Naqvi H, Mahdi F - Iran J Cancer Prev (2012)

Bottom Line: Mutations at codons Ile571Leu and Trp582Ser was found in two independent cases.The presence of c-kit mutations in our study adds to investigative spectrum of AML cases.Further studies in larger group of cases possibly will be required to determine the prognostic implications and to investigate how these mutations are co-related to the progression and pathogenesis of AML.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Biochemistry, Era's Lucknow Medical College and Hospital, Lucknow, India.

ABSTRACT

Background: Acute Myeloid Leukaemia (AML) is a cancer of blood-forming cells in bone marrow. C-kit gene is a Receptor Tyrosine Kinase class III (RTK) that is expressed by early hematopoietic progenitor cells and plays an important role in hematopoietic stem cell proliferation, differentiation and survival. It is known that c-kit is a proto-oncogene and the activating c-kit mutations are likely to contribute in the development of leukaemia in humans. Exon 11 of c-Kit gene is the frequent site for mutations in different kinds of tumours.

Methods: In order to determine the frequency and prevalence of exon 11 mutations in 51 AML cases, we have done polymerase chain reaction-single-strand conformational polymorphism followed by direct DNA sequencing.

Results: The c-kit mutations in exon 11 were detected in 15.68% (8/51) in AML cases. We have detected totally ten missense mutations in eight AML cases those include Lys550Asn, Tyr568Ser, Ile571Leu, Tyr578Pro, Trp582Ser and Arg588Met and novel missense mutations at codons Ile563Lys and Val569Leu. Mutations at codons Ile571Leu and Trp582Ser was found in two independent cases.

Conclusion: The presence of c-kit mutations in our study adds to investigative spectrum of AML cases. Since the c-kit mutations are seen in other malignancies, mutations in exon 11 of the c-kit gene might be involve in pathogenesis and represent useful predictive genetic marker in AML. Further studies in larger group of cases possibly will be required to determine the prognostic implications and to investigate how these mutations are co-related to the progression and pathogenesis of AML.

No MeSH data available.


Related in: MedlinePlus

Sizing of 257bp long PCR products of c-kit exon 11 on 2 % agarose in AML cases. 50bp Ladder in lane 4th and cases in 1, 2, 3, 5 and 6th lane.
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f1-IJCP-05-027: Sizing of 257bp long PCR products of c-kit exon 11 on 2 % agarose in AML cases. 50bp Ladder in lane 4th and cases in 1, 2, 3, 5 and 6th lane.

Mentions: Polymerase Chain Reaction (PCR) was performed with 25μl PCR reaction mixture containing 200ng of template DNA, 10 pmol of each primer, 10mmol/L of each mix dNTPs, 1X reaction buffer and 0.3 units of Taq polymerase enzyme (Fermentas, Germany) in an MJ Mini Thermocycler (Bio-Rad, UK). As per the cycling conditions denaturation at 94°C for 30 seconds, followed by annealing at 56°C for 30 seconds, and extension at 72°C for 30 seconds, repeated for 35 cycles followed by a final extension step at 72°C for 10 minutes using the primers [11], forward 5- ATTATTAAAAGGTGATCTATTTTTC- 3 and reverse 5- ACTGTTATGTGTACCCAAAAAG- 3. Resultantly, obtained 257 bp long amplicons and tested out on 2% agarose gel electrophoresis (Figure1).


Screening of C-kit gene Mutation in Acute Myeloid Leukaemia in Northern India.

Hussain SR, Raza ST, Babu SG, Singh P, Naqvi H, Mahdi F - Iran J Cancer Prev (2012)

Sizing of 257bp long PCR products of c-kit exon 11 on 2 % agarose in AML cases. 50bp Ladder in lane 4th and cases in 1, 2, 3, 5 and 6th lane.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4352523&req=5

f1-IJCP-05-027: Sizing of 257bp long PCR products of c-kit exon 11 on 2 % agarose in AML cases. 50bp Ladder in lane 4th and cases in 1, 2, 3, 5 and 6th lane.
Mentions: Polymerase Chain Reaction (PCR) was performed with 25μl PCR reaction mixture containing 200ng of template DNA, 10 pmol of each primer, 10mmol/L of each mix dNTPs, 1X reaction buffer and 0.3 units of Taq polymerase enzyme (Fermentas, Germany) in an MJ Mini Thermocycler (Bio-Rad, UK). As per the cycling conditions denaturation at 94°C for 30 seconds, followed by annealing at 56°C for 30 seconds, and extension at 72°C for 30 seconds, repeated for 35 cycles followed by a final extension step at 72°C for 10 minutes using the primers [11], forward 5- ATTATTAAAAGGTGATCTATTTTTC- 3 and reverse 5- ACTGTTATGTGTACCCAAAAAG- 3. Resultantly, obtained 257 bp long amplicons and tested out on 2% agarose gel electrophoresis (Figure1).

Bottom Line: Mutations at codons Ile571Leu and Trp582Ser was found in two independent cases.The presence of c-kit mutations in our study adds to investigative spectrum of AML cases.Further studies in larger group of cases possibly will be required to determine the prognostic implications and to investigate how these mutations are co-related to the progression and pathogenesis of AML.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Biochemistry, Era's Lucknow Medical College and Hospital, Lucknow, India.

ABSTRACT

Background: Acute Myeloid Leukaemia (AML) is a cancer of blood-forming cells in bone marrow. C-kit gene is a Receptor Tyrosine Kinase class III (RTK) that is expressed by early hematopoietic progenitor cells and plays an important role in hematopoietic stem cell proliferation, differentiation and survival. It is known that c-kit is a proto-oncogene and the activating c-kit mutations are likely to contribute in the development of leukaemia in humans. Exon 11 of c-Kit gene is the frequent site for mutations in different kinds of tumours.

Methods: In order to determine the frequency and prevalence of exon 11 mutations in 51 AML cases, we have done polymerase chain reaction-single-strand conformational polymorphism followed by direct DNA sequencing.

Results: The c-kit mutations in exon 11 were detected in 15.68% (8/51) in AML cases. We have detected totally ten missense mutations in eight AML cases those include Lys550Asn, Tyr568Ser, Ile571Leu, Tyr578Pro, Trp582Ser and Arg588Met and novel missense mutations at codons Ile563Lys and Val569Leu. Mutations at codons Ile571Leu and Trp582Ser was found in two independent cases.

Conclusion: The presence of c-kit mutations in our study adds to investigative spectrum of AML cases. Since the c-kit mutations are seen in other malignancies, mutations in exon 11 of the c-kit gene might be involve in pathogenesis and represent useful predictive genetic marker in AML. Further studies in larger group of cases possibly will be required to determine the prognostic implications and to investigate how these mutations are co-related to the progression and pathogenesis of AML.

No MeSH data available.


Related in: MedlinePlus