PHA-Induced Peripheral Blood Cytogenetics and Molecular Analysis: a Valid Diagnostic and Follow-up Modality for Acute Promyelocytic Leukemia Patients Treated with ATRA and/or Arsenic Tri-oxide

Asian Pac J Cancer Prev. 2016;17(4):1999-2006. doi: 10.7314/apjcp.2016.17.4.1999.

Abstract

Background: Acute promyelocytic leukemia (APML) is characterized by the reciprocal translocation t(15;17) (q22;q12) resulting in the PML-RARα fusion gene. A dual diagnostic and follow up approach was applied including cytogenetic demonstration of the t(15;17) translocation and detection of PML-RARα chimeric transcripts by molecular means.

Purpose: Conventional cytogenetics involving bone marrow is beset with high probability of poor metaphase index and was substituted with phytohemagglutinin (PHA)-induced peripheral blood culture based cytogenetic analysis as a diagnostic and follow up modality in APML patients of Kashmir (North India). Both qualitative (RT-PCR) and quantitative (Q-PCR) tests were simultaneously carried out to authenticate the modified cytogenetics.

Materials and methods: Patient samples were subjected to the said techniques to establish their baseline as well as follow-up status.

Results: Initial cytogenetics revealed 30 patients (81%) positive for t(15;17) whereas 7 (19%) had either cryptic translocation or were negative for t(15;17). Two cases had chromosome 16q deletion and no hallmark translocation t(15;17). Q-PCR status for PML-RARα was found to be positive for all patients. All the APML patients were reassessed at the end of consolidation phase and during maintenance phase of chemotherapy where 6 patients had molecular relapse, wherein 4 also demonstrated cytogenetic relapse.

Conclusions: It was found that PHA-induced peripheral blood cytogenetics along with molecular analysis could prove a reliable modality in the diagnosis and assessment of follow up response of APML patients.

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Antineoplastic Combined Chemotherapy Protocols / therapeutic use*
  • Arsenic Trioxide
  • Arsenicals / administration & dosage
  • Biomarkers, Tumor / genetics
  • Blood Cells / metabolism
  • Blood Cells / pathology*
  • Child
  • Chromosomes, Human, Pair 15 / genetics
  • Chromosomes, Human, Pair 17 / genetics
  • Cytogenetic Analysis
  • Female
  • Follow-Up Studies
  • Gene Rearrangement
  • Humans
  • In Situ Hybridization, Fluorescence
  • India
  • Leukemia, Promyelocytic, Acute / blood
  • Leukemia, Promyelocytic, Acute / diagnosis*
  • Leukemia, Promyelocytic, Acute / drug therapy
  • Leukemia, Promyelocytic, Acute / genetics
  • Male
  • Middle Aged
  • Neoplasm Recurrence, Local / drug therapy
  • Neoplasm Recurrence, Local / pathology
  • Neoplasm Staging
  • Oncogene Proteins, Fusion / genetics
  • Oxides / administration & dosage
  • Phytohemagglutinins / pharmacology
  • Prognosis
  • Translocation, Genetic
  • Tretinoin / administration & dosage

Substances

  • Arsenicals
  • Biomarkers, Tumor
  • Oncogene Proteins, Fusion
  • Oxides
  • Phytohemagglutinins
  • promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein
  • Tretinoin
  • Arsenic Trioxide