No evidence for amplification of V617F JAK2 in myeloproliferative disorders

Selective reduction of JAK2V617F-dependent cell growth by siRNA/shRNA and its reversal by cytokines

“…The HEL cells are homozygous for JAK2 V617F and carry an estimated 8 copies of the JAK2 V617F gene per cell. 24,25 In contrast, UT-7 cells are homozygous for the WT gene. Transfection of HEL cells with the mt1-and mt4-siRNAs, which include the mutation, resulted in a 55% (Ϯ 6%; n ϭ 3) decrease in JAK2 protein expression in nonselected cell population compared with nontreated cells ( Figure 1B).…”

“…24 The SET-2 cells carry 5 JAK2 copies probably including 4 mutated JAK2 V617F . 25 All these cells were infected with the pRRL lentiviruses carrying the shRNAs, and transduced cells were sorted based on GFP expression. Western blot analysis revealed that either pRRL-mt1 or pRRL-mt4 reduced the expression of the JAK2 V617F protein in HEL (54% or 77%, respectively) and UKE-1 (56% or 78%, respectively) cells ( Figure 2C).…”

Identification of myeloproliferative neoplasm drug agents via predictive simulation modeling: assessing responsiveness with micro-environment derived cytokines

“…The SET2 and HEL cell line models that are representative of Jak2-V617F driven MPN were created as described in the methods. The mutation and copy number variation (CNV) aberration data for these cell lines were obtained from the literature and cancer portals such as Sanger and cBioportal [14–16, Appendix 1]. SET2 cells, besides having the Jak2-V617F mutation, also has mutations in RPTOR, TP53, CCNA2, NOTCH2, EGF, MAP2K1 as well as multiple gene amplifications and deletions as listed in Appendix 1.…”

“…To test this, we selected two representative MPN cell lines, Human erythroleukemia 92.1.7 (HEL) and SET2, and obtained their genomic profile. The HEL cell is largely defined by the Jak2-V617F mutation, along with deletion of CDKN2A, RB1 and TP53 [14–16]; while the SET-2 cells are defined by Jak2 mutations as well as TP53, and additional mutations and copy number variations as reported in C-BioPortal [16]. Simulation avatars were then created for each cell line and the signaling networks were simulated, both with and without inflammatory cytokines tumor necrosis factor alpha (TNFα), interferon gamma (IFNγ) and interleukin 6 (IL-6).…”