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CXXC5 (retinoid-inducible nuclear factor, RINF) is a potential therapeutic target in high-risk human acute myeloid leukemia.

Astori A, Fredly H, Aloysius TA, Bullinger L, Mansat-De Mas V, de la Grange P, Delhommeau F, Hagen KM, Récher C, Dusanter-Fourt I, Knappskog S, Lillehaug JR, Pendino F, Bruserud Ø - Oncotarget (2013)

Bottom Line: Furthermore, patients with low-risk cytogenetic abnormalities showed significantly lower levels compared to patients with high-risk abnormalities, and high RINF/CXXC5/ mRNA levels were associated with decreased overall survival for patients receiving intensive chemotherapy for newly diagnosed AML.This association with prognosis was seen both when investigating (i) an unselected patient population as well as for patients with (ii) normal cytogenetic and (iii) core-binding factor AML.The association with adverse prognosis together with the antiapoptotic effect of CXXC5/RINF suggests that targeting of CXXC5/RINF should be considered as a possible therapeutic strategy, especially in high-risk patients who show increased expression in AML cells compared with normal hematopoietic cells.

View Article: PubMed Central - PubMed

Affiliation: Inserm, U1016, Institut Cochin, F-75014, Paris, France.

ABSTRACT
The retinoid-responsive gene CXXC5 localizes to the 5q31.2 chromosomal region and encodes a retinoid-inducible nuclear factor (RINF) that seems important during normal myelopoiesis. We investigated CXXC5/RINF expression in primary human acute myeloid leukemia (AML) cells derived from 594 patients, and a wide variation in CXXC5/RINF mRNA levels was observed both in the immature leukemic myeloblasts and in immature acute lymphoblastic leukemia cells. Furthermore, patients with low-risk cytogenetic abnormalities showed significantly lower levels compared to patients with high-risk abnormalities, and high RINF/CXXC5/ mRNA levels were associated with decreased overall survival for patients receiving intensive chemotherapy for newly diagnosed AML. This association with prognosis was seen both when investigating (i) an unselected patient population as well as for patients with (ii) normal cytogenetic and (iii) core-binding factor AML. CXXC5/RINF knockdown in AML cell lines caused increased susceptibility to chemotherapy-induced apoptosis, and regulation of apoptosis also seemed to differ between primary human AML cells with high and low RINF expression. The association with adverse prognosis together with the antiapoptotic effect of CXXC5/RINF suggests that targeting of CXXC5/RINF should be considered as a possible therapeutic strategy, especially in high-risk patients who show increased expression in AML cells compared with normal hematopoietic cells.

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RINF knock-down sensitizes K562 cells to doxorubicin- and lenalidomid-induced apoptosisK562 cells stably expressing shRNA/RINF or a control shRNA/scramble were examined. (A) RINF expression; shRNA/RINF was associated with decreased RINF expression both at the mRNA and protein level. (B) Effect of RINF knockdown on AML cell proliferation, RINF knockdown did not alter K562 proliferation during 7 days of in vitro culture. (C, D) K562 cells were cultured for 4 days with 50 μM of lenalidomide or 200 ng/μl of doxorubicin. Cell viability and cell death was then assessed at the same time. (C) WST1-cell viability and proliferation assay or (D) TUNEL assay was realized before a DAPI staining. (D) Morphology was evaluated by light microscopy of May-Grünwald-Giemsa stained cytospin smears. The observations are consistent with an increased susceptibility to chemotherapy-induced apoptosis after RINF knockdown.
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Figure 3: RINF knock-down sensitizes K562 cells to doxorubicin- and lenalidomid-induced apoptosisK562 cells stably expressing shRNA/RINF or a control shRNA/scramble were examined. (A) RINF expression; shRNA/RINF was associated with decreased RINF expression both at the mRNA and protein level. (B) Effect of RINF knockdown on AML cell proliferation, RINF knockdown did not alter K562 proliferation during 7 days of in vitro culture. (C, D) K562 cells were cultured for 4 days with 50 μM of lenalidomide or 200 ng/μl of doxorubicin. Cell viability and cell death was then assessed at the same time. (C) WST1-cell viability and proliferation assay or (D) TUNEL assay was realized before a DAPI staining. (D) Morphology was evaluated by light microscopy of May-Grünwald-Giemsa stained cytospin smears. The observations are consistent with an increased susceptibility to chemotherapy-induced apoptosis after RINF knockdown.

Mentions: To further investigate whether RINF contributes to the chemoresistant phenotype of high-risk patients we investigated the effect of RINF knockdown on the susceptibility of five different immature myeloid leukemic cell lines to chemotherapy-induced apoptosis, including K562, MV4-11, OCI-AML3, NB4 and HL60. The highest RINF mRNA levels were observed for K562 and MV4-11 cells, and these cells showed an increased susceptibility to drug-induced apoptosis after RINF silencing. The results for K562 are presented in detail in Fig. 3. An efficient knock-down of RINF at the protein level was documented (Fig. 3A), but the in vitro proliferation was not altered (Fig. 3B) and apoptosis was minimal during in vitro culture in the absence of chemotherapy (Fig. 3C, left). However, RINF knockdown was associated with increased sensitivity to daunorubicin- and lenalidomide-induced apoptosis (Figs. 3C and 3D). The susceptibility to drug-induced apoptosis was not affected by RINF silencing for the three cell lines with low expression (data not shown). All these effects were reproduced in independent experiments.


CXXC5 (retinoid-inducible nuclear factor, RINF) is a potential therapeutic target in high-risk human acute myeloid leukemia.

Astori A, Fredly H, Aloysius TA, Bullinger L, Mansat-De Mas V, de la Grange P, Delhommeau F, Hagen KM, Récher C, Dusanter-Fourt I, Knappskog S, Lillehaug JR, Pendino F, Bruserud Ø - Oncotarget (2013)

RINF knock-down sensitizes K562 cells to doxorubicin- and lenalidomid-induced apoptosisK562 cells stably expressing shRNA/RINF or a control shRNA/scramble were examined. (A) RINF expression; shRNA/RINF was associated with decreased RINF expression both at the mRNA and protein level. (B) Effect of RINF knockdown on AML cell proliferation, RINF knockdown did not alter K562 proliferation during 7 days of in vitro culture. (C, D) K562 cells were cultured for 4 days with 50 μM of lenalidomide or 200 ng/μl of doxorubicin. Cell viability and cell death was then assessed at the same time. (C) WST1-cell viability and proliferation assay or (D) TUNEL assay was realized before a DAPI staining. (D) Morphology was evaluated by light microscopy of May-Grünwald-Giemsa stained cytospin smears. The observations are consistent with an increased susceptibility to chemotherapy-induced apoptosis after RINF knockdown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3824541&req=5

Figure 3: RINF knock-down sensitizes K562 cells to doxorubicin- and lenalidomid-induced apoptosisK562 cells stably expressing shRNA/RINF or a control shRNA/scramble were examined. (A) RINF expression; shRNA/RINF was associated with decreased RINF expression both at the mRNA and protein level. (B) Effect of RINF knockdown on AML cell proliferation, RINF knockdown did not alter K562 proliferation during 7 days of in vitro culture. (C, D) K562 cells were cultured for 4 days with 50 μM of lenalidomide or 200 ng/μl of doxorubicin. Cell viability and cell death was then assessed at the same time. (C) WST1-cell viability and proliferation assay or (D) TUNEL assay was realized before a DAPI staining. (D) Morphology was evaluated by light microscopy of May-Grünwald-Giemsa stained cytospin smears. The observations are consistent with an increased susceptibility to chemotherapy-induced apoptosis after RINF knockdown.
Mentions: To further investigate whether RINF contributes to the chemoresistant phenotype of high-risk patients we investigated the effect of RINF knockdown on the susceptibility of five different immature myeloid leukemic cell lines to chemotherapy-induced apoptosis, including K562, MV4-11, OCI-AML3, NB4 and HL60. The highest RINF mRNA levels were observed for K562 and MV4-11 cells, and these cells showed an increased susceptibility to drug-induced apoptosis after RINF silencing. The results for K562 are presented in detail in Fig. 3. An efficient knock-down of RINF at the protein level was documented (Fig. 3A), but the in vitro proliferation was not altered (Fig. 3B) and apoptosis was minimal during in vitro culture in the absence of chemotherapy (Fig. 3C, left). However, RINF knockdown was associated with increased sensitivity to daunorubicin- and lenalidomide-induced apoptosis (Figs. 3C and 3D). The susceptibility to drug-induced apoptosis was not affected by RINF silencing for the three cell lines with low expression (data not shown). All these effects were reproduced in independent experiments.

Bottom Line: Furthermore, patients with low-risk cytogenetic abnormalities showed significantly lower levels compared to patients with high-risk abnormalities, and high RINF/CXXC5/ mRNA levels were associated with decreased overall survival for patients receiving intensive chemotherapy for newly diagnosed AML.This association with prognosis was seen both when investigating (i) an unselected patient population as well as for patients with (ii) normal cytogenetic and (iii) core-binding factor AML.The association with adverse prognosis together with the antiapoptotic effect of CXXC5/RINF suggests that targeting of CXXC5/RINF should be considered as a possible therapeutic strategy, especially in high-risk patients who show increased expression in AML cells compared with normal hematopoietic cells.

View Article: PubMed Central - PubMed

Affiliation: Inserm, U1016, Institut Cochin, F-75014, Paris, France.

ABSTRACT
The retinoid-responsive gene CXXC5 localizes to the 5q31.2 chromosomal region and encodes a retinoid-inducible nuclear factor (RINF) that seems important during normal myelopoiesis. We investigated CXXC5/RINF expression in primary human acute myeloid leukemia (AML) cells derived from 594 patients, and a wide variation in CXXC5/RINF mRNA levels was observed both in the immature leukemic myeloblasts and in immature acute lymphoblastic leukemia cells. Furthermore, patients with low-risk cytogenetic abnormalities showed significantly lower levels compared to patients with high-risk abnormalities, and high RINF/CXXC5/ mRNA levels were associated with decreased overall survival for patients receiving intensive chemotherapy for newly diagnosed AML. This association with prognosis was seen both when investigating (i) an unselected patient population as well as for patients with (ii) normal cytogenetic and (iii) core-binding factor AML. CXXC5/RINF knockdown in AML cell lines caused increased susceptibility to chemotherapy-induced apoptosis, and regulation of apoptosis also seemed to differ between primary human AML cells with high and low RINF expression. The association with adverse prognosis together with the antiapoptotic effect of CXXC5/RINF suggests that targeting of CXXC5/RINF should be considered as a possible therapeutic strategy, especially in high-risk patients who show increased expression in AML cells compared with normal hematopoietic cells.

Show MeSH
Related in: MedlinePlus