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MLL-SEPT5 Fusion Transcript in Two de novo Acute Myeloid Leukemia Patients With t(11;22)(q23;q11)

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Dear Editor, Rearrangements involving mixed lineage leukemia (MLL) are common chromosome aberrations in infant, pediatric and adult acute leukemia, which are generally associated with poor prognosis... Total RNA was extracted from the bone marrow cells using TRIzol reagent (Invitrogen, Paisley, UK) and chloroform. cDNA was obtained by RT-PCR using the M-MLV reverse transcriptase (Promega, Madison, WI, USA)... The resulting cDNA was amplified by PCR using 2×Hieff PCR Master Mix (Yaeson, Shanghai, China) and the following primers: 5'-GCTCCACCCATCAAACCAAT-3' from exon 5 of MLL and 5'-TTCTTCTCAATGTCCACCGT-3' from exon 4 of SEPT5 (Fig. 2)... Karyotype was 46, XX, t(11;22)(q23;q11.2) in all 10 cells examined... She did not receive chemotherapy in our hospital... MLL-SEPT5 fusion was verified by the previously used method and the following PCR primers: 5'-CCGGTCAATAAGCAGGAGAA-3' from exon 10 of MLL and 5'-CAGCCATGAGTGTGAAG-3' from exon 3 of SEPT5... Long distance inverse-PCR is very useful for recognizing unusual cryptic cytogenetic findings, such as a rare MLL partner gene, or discrepancies in molecular results that are hard to identify through common molecular diagnostic methods... Since we have identified the partner gene from previous studies, we utilized RT-PCR and Sanger DNA sequencing to confirm the fusion gene... The results indicate the rearrangement between MLL exon 10 and SEPT5 exon 3 (Fig. 3B)... We described two new cases of de novo AML with t(11;22)(q23;q11.2), resulting in a MLL-SEPT5 fusion... All patients with the MLL-SEPT5 fusion gene were diagnosed with AML... Tatsumi et al. examined SEPT5 expression in 27 leukemia cell lines, including 13 AML and 14 ALL cell lines, which indicated that SEPT5 expression in AML cell lines was significantly higher than that in ALL... MLL-SEPT5 fusion gene was important in the leukemogenesis of AML with t(11;22)(q23;q11.2).

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Identification of the MLL-SEPT5 fusion transcripts by reverse transcription-PCR. Lane M, 1kb DNA ladder; Lane E, empty control; Lane N, PCR control; Lane P1, patient 1; Lane P2, patient 2.Abbreviation: MLL, mixed lineage leukemia.
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Figure 2: Identification of the MLL-SEPT5 fusion transcripts by reverse transcription-PCR. Lane M, 1kb DNA ladder; Lane E, empty control; Lane N, PCR control; Lane P1, patient 1; Lane P2, patient 2.Abbreviation: MLL, mixed lineage leukemia.

Mentions: To validate MLL-SEPT5 existence, we performed RT-PCR and direct DNA sequencing. Total RNA was extracted from the bone marrow cells using TRIzol reagent (Invitrogen, Paisley, UK) and chloroform. cDNA was obtained by RT-PCR using the M-MLV reverse transcriptase (Promega, Madison, WI, USA). The resulting cDNA was amplified by PCR using 2×Hieff PCR Master Mix (Yaeson, Shanghai, China) and the following primers: 5'-GCTCCACCCATCAAACCAAT-3' from exon 5 of MLL and 5'-TTCTTCTCAATGTCCACCGT-3' from exon 4 of SEPT5 (Fig. 2). PCR products were sequenced on both strands by using an Applied Biosystems ABI 3730 XL DNA analyzer (Thermo Fisher Scientific, Waltham, MA, USA). The sequence was analyzed with Chromas Lite software (version 2.01, Technelysium, Brisbane, Australia). The fusion between MLL exon 8 and SEPT5 intron 2 is indicated by the chromatogram (Fig. 3A). Both breakpoints in the two genes have not been reported previously.


MLL-SEPT5 Fusion Transcript in Two de novo Acute Myeloid Leukemia Patients With t(11;22)(q23;q11)
Identification of the MLL-SEPT5 fusion transcripts by reverse transcription-PCR. Lane M, 1kb DNA ladder; Lane E, empty control; Lane N, PCR control; Lane P1, patient 1; Lane P2, patient 2.Abbreviation: MLL, mixed lineage leukemia.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940498&req=5

Figure 2: Identification of the MLL-SEPT5 fusion transcripts by reverse transcription-PCR. Lane M, 1kb DNA ladder; Lane E, empty control; Lane N, PCR control; Lane P1, patient 1; Lane P2, patient 2.Abbreviation: MLL, mixed lineage leukemia.
Mentions: To validate MLL-SEPT5 existence, we performed RT-PCR and direct DNA sequencing. Total RNA was extracted from the bone marrow cells using TRIzol reagent (Invitrogen, Paisley, UK) and chloroform. cDNA was obtained by RT-PCR using the M-MLV reverse transcriptase (Promega, Madison, WI, USA). The resulting cDNA was amplified by PCR using 2×Hieff PCR Master Mix (Yaeson, Shanghai, China) and the following primers: 5'-GCTCCACCCATCAAACCAAT-3' from exon 5 of MLL and 5'-TTCTTCTCAATGTCCACCGT-3' from exon 4 of SEPT5 (Fig. 2). PCR products were sequenced on both strands by using an Applied Biosystems ABI 3730 XL DNA analyzer (Thermo Fisher Scientific, Waltham, MA, USA). The sequence was analyzed with Chromas Lite software (version 2.01, Technelysium, Brisbane, Australia). The fusion between MLL exon 8 and SEPT5 intron 2 is indicated by the chromatogram (Fig. 3A). Both breakpoints in the two genes have not been reported previously.

View Article: PubMed Central - PubMed

AUTOMATICALLY GENERATED EXCERPT
Please rate it.

Dear Editor, Rearrangements involving mixed lineage leukemia (MLL) are common chromosome aberrations in infant, pediatric and adult acute leukemia, which are generally associated with poor prognosis... Total RNA was extracted from the bone marrow cells using TRIzol reagent (Invitrogen, Paisley, UK) and chloroform. cDNA was obtained by RT-PCR using the M-MLV reverse transcriptase (Promega, Madison, WI, USA)... The resulting cDNA was amplified by PCR using 2×Hieff PCR Master Mix (Yaeson, Shanghai, China) and the following primers: 5'-GCTCCACCCATCAAACCAAT-3' from exon 5 of MLL and 5'-TTCTTCTCAATGTCCACCGT-3' from exon 4 of SEPT5 (Fig. 2)... Karyotype was 46, XX, t(11;22)(q23;q11.2) in all 10 cells examined... She did not receive chemotherapy in our hospital... MLL-SEPT5 fusion was verified by the previously used method and the following PCR primers: 5'-CCGGTCAATAAGCAGGAGAA-3' from exon 10 of MLL and 5'-CAGCCATGAGTGTGAAG-3' from exon 3 of SEPT5... Long distance inverse-PCR is very useful for recognizing unusual cryptic cytogenetic findings, such as a rare MLL partner gene, or discrepancies in molecular results that are hard to identify through common molecular diagnostic methods... Since we have identified the partner gene from previous studies, we utilized RT-PCR and Sanger DNA sequencing to confirm the fusion gene... The results indicate the rearrangement between MLL exon 10 and SEPT5 exon 3 (Fig. 3B)... We described two new cases of de novo AML with t(11;22)(q23;q11.2), resulting in a MLL-SEPT5 fusion... All patients with the MLL-SEPT5 fusion gene were diagnosed with AML... Tatsumi et al. examined SEPT5 expression in 27 leukemia cell lines, including 13 AML and 14 ALL cell lines, which indicated that SEPT5 expression in AML cell lines was significantly higher than that in ALL... MLL-SEPT5 fusion gene was important in the leukemogenesis of AML with t(11;22)(q23;q11.2).

No MeSH data available.


Related in: MedlinePlus