Biochemical differences between SUDHL-1 and KARPAS 299 cells derived from t(2;5)-positive anaplastic large cell lymphoma are responsible for the different sensitivity to the antiproliferative effect of p27Kip1

Abstract

An inverse correlation between p27 Kip1 expression and proliferation has been recently established in tissues derived from human lymphomas. The nucleophosminanaplastic lymphoma kinase (NPM ± ALK)/phospholipase C-g (PLCg) complex also appears to play an important role in cell proliferation and malignant transformation of anaplastic large cell lymphoma (ALCL). In this study, we report that SUDHL-1 and KARPAS 299 ALCL-derived cell lines present di erent sensitivity to the antiproliferative e ect of recombinant adenovirus-mediated p27 Kip1 expression or to serumstarvation in culture media. The results indicate that exogenous p27 Kip1 may interact with the NPM ± ALK/ PLCg pathway in SUDHL-1 but not in KARPAS 299 cells. This interaction correlates with changes in cell cycle and cell morphology observed mainly in SUDHL-1 cells. The percentage of SUDHL-1 cells in S phase declines, whereas it is almost unchanged in KARPAS 299 cells as compared to the controls after 96 h of infection with the recombinant adenovirus. Furthermore KARPAS 299 cells are resistant to serum-starvation due to de®cient p27 Kip1 -upregulation and G1 arrest, whereas SUDHL-1 cells respond with increased G1 phase and p27 Kip1 -upregulation after 48 h of serum-starvation. Both cell lines express appropriate variation of levels of cyclins E and A, and Rb-phosphorylation as expected by growing them in culture media with di erent FBS content. Although both cell lines express cyclin D2, SUDHL-1 cells only present high level of cyclin D3. Moreover SUDHL-1 cells express high level of PTEN and the PKB/Akt pathway is constitutively activated in both cell lines. Lastly SUDHL-1 cells show higher levels of phosphotyrosine-containing proteins that is correlated with a higher NPM ± ALK-associated autophosphorylation activity compared to KARPAS 299 cells. Our study clearly identi®es some of the biochemical di erences that may explain the di erence in sensitivity to antiproliferative stimuli shown by two cell lines derived from the same type of lymphoma. Oncogene (2001) 20, 4466 ± 4475.

Recombinant adenovirus-mediated cytotoxic gene therapy of lymphoproliferative disorders: is CAR important for the vector to ride?

“…[17][18][19] For the first time, we showed that adenovirus-mediated p27 Kip1 expression was able to induce in vitro apoptosis in SUDHL-1 cells derived from t(2:5)-ALCL. 20, 21 We also demonstrated that the effects of adenovirus-mediated p27 Kip1 expression were cell dependent and differed from the effects of adenovirus-mediated p53 expression in SUDHL-1 cells. 20 22 On the contrary, cells derived from HD were resistant to apoptosis induced by the same recombinant adenoviruses.…”

“…20, 21 We also demonstrated that the effects of adenovirus-mediated p27 Kip1 expression were cell dependent and differed from the effects of adenovirus-mediated p53 expression in SUDHL-1 cells. 20 22 On the contrary, cells derived from HD were resistant to apoptosis induced by the same recombinant adenoviruses. 22 The difference in the level of apoptosis was correlated with the level of phosphorylation of the retinoblastoma (Rb) protein in the cell lines tested.…”

“…12,15,[20][21][22] In one study, we showed that the recombinant adenovirusmediated expression of exogenous p27 Kip1 was equally efficient in the cytoplasm and in the nucleus of SUDHL-1 and KARPAS 299 cells derived from t(2;5)-ALCL. 20 The presence of CAR and integrins on the cell surface of SUDHL-1 (Ref. 12) and KARPAS 299 cells (unpublished data) and the efficiency of the CMV promoter allowed the recombinant adenovirus Adp27 to express the cytotoxic gene.…”

“…12) and KARPAS 299 cells (unpublished data) and the efficiency of the CMV promoter allowed the recombinant adenovirus Adp27 to express the cytotoxic gene. 20 When we assessed the consequences of the expression of p27 Kip1 on the life of the two cell lines, there were differences in response between the two cell lines tested. Biochemical differences in the signaling pathways between the two cell lines were responsible for the different response to the same recombinant adenovirus.…”

TrkA is a binding partner of NPM‐ALK that promotes the survival of ALK+ T‐cell lymphoma

“…Notably, some variability in the cellular response to TrkAi was observed among the different lymphoma cell lines. This variability could be explained, at least in part, by the intrinsic biochemical differences among these cell lines (Turturro et al ., 2001). Importantly, the cellular and biochemical effects of selective inhibition of TrkA by TrkAi were consistent when specific downregulation of TrkA was achieved by using siRNA.…”