Limits...
Molecular targeting of the oncoprotein PLK1 in pediatric acute myeloid leukemia: RO3280, a novel PLK1 inhibitor, induces apoptosis in leukemia cells.

Wang NN, Li ZH, Zhao H, Tao YF, Xu LX, Lu J, Cao L, Du XJ, Sun LC, Zhao WL, Xiao PF, Fang F, Su GH, Li YH, Li G, Li YP, Xu YY, Zhou HT, Wu Y, Jin MF, Liu L, Ni J, Wang J, Hu SY, Zhu XM, Feng X, Pan J - Int J Mol Sci (2015)

Bottom Line: PLK1 mRNA expression was significantly higher in AML samples compared with control samples (82.95 ± 110.28 vs. 6.36 ± 6.35; p < 0.001).RO3280 treatment regulated several apoptosis-associated genes.These results provide insights into the potential use of RO3280 for AML therapy; however, the underlying mechanisms remain to be determined.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou 215003, China. wangnn90s@163.com.

ABSTRACT
Polo-like kinase 1 (PLK1) is highly expressed in many cancers and therefore a biomarker of transformation and potential target for the development of cancer-specific small molecule drugs. RO3280 was recently identified as a novel PLK1 inhibitor; however its therapeutic effects in leukemia treatment are still unknown. We found that the PLK1 protein was highly expressed in leukemia cell lines as well as 73.3% (11/15) of pediatric acute myeloid leukemia (AML) samples. PLK1 mRNA expression was significantly higher in AML samples compared with control samples (82.95 ± 110.28 vs. 6.36 ± 6.35; p < 0.001). Kaplan-Meier survival analysis revealed that shorter survival time correlated with high tumor PLK1 expression (p = 0.002). The 50% inhibitory concentration (IC50) of RO3280 for acute leukemia cells was between 74 and 797 nM. The IC50 of RO3280 in primary acute lymphocytic leukemia (ALL) and AML cells was between 35.49 and 110.76 nM and 52.80 and 147.50 nM, respectively. RO3280 induced apoptosis and cell cycle disorder in leukemia cells. RO3280 treatment regulated several apoptosis-associated genes. The regulation of DCC, CDKN1A, BTK, and SOCS2 was verified by western blot. These results provide insights into the potential use of RO3280 for AML therapy; however, the underlying mechanisms remain to be determined.

Show MeSH

Related in: MedlinePlus

Western blot verification of real-time PCR array results. Western blot analysis of cells following a 24 h treatment with RO3280 at 50 or 100 nM compared with DMSO control mock treatment. The upregulation of DCC and CDKN1A and down regulation of BTK and SOCS2 in RO3280 treated cells. Protein lysates from treated cells were tested for expression levels by Western blot analysis.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4307303&req=5

ijms-16-01266-f007: Western blot verification of real-time PCR array results. Western blot analysis of cells following a 24 h treatment with RO3280 at 50 or 100 nM compared with DMSO control mock treatment. The upregulation of DCC and CDKN1A and down regulation of BTK and SOCS2 in RO3280 treated cells. Protein lysates from treated cells were tested for expression levels by Western blot analysis.

Mentions: In order to identify apoptosis and/or programmed cell death molecules implicated in the effects of RO3280, we used the SABioscience Human Apoptosis PCR Array PAHS-3012. With this real-time PCR array we analyzed and clustered the expression of 370 genes associated with apoptosis in DMSO or RO3280 treated cells (Figure 6A). The genes most significantly upregulated or downregulated are shown in Figure 6B,C, respectively. Examination of the array data revealed that 32 genes were significantly upregulated and 16 genes were significantly downregulated in RO3280 treatment group compared with DMSO control group (Table 6 and Table 7, respectively). The upregulated genes included protein kinase C zeta, receptor-interacting serine-threonine kinase 3, harakiri BCL2 interacting protein, DCC netrin 1 receptor, and cyclin-dependent kinase inhibitor 1A. The downregulated genes included interleukin 1α, nucleotide-binding oligomerization domain containing 2, caspase-1, apoptosis-related cysteine peptidase, serpin peptidase inhibitor clade B, B-cell CLL/lymphoma 2, and Bruton agammaglobulinemia tyrosine kinase. The RO3280-dependent upregulation of DCC and CDKN1A and downregulation of BTK and SOCS2 was verified by western blot analysis (Figure 7).


Molecular targeting of the oncoprotein PLK1 in pediatric acute myeloid leukemia: RO3280, a novel PLK1 inhibitor, induces apoptosis in leukemia cells.

Wang NN, Li ZH, Zhao H, Tao YF, Xu LX, Lu J, Cao L, Du XJ, Sun LC, Zhao WL, Xiao PF, Fang F, Su GH, Li YH, Li G, Li YP, Xu YY, Zhou HT, Wu Y, Jin MF, Liu L, Ni J, Wang J, Hu SY, Zhu XM, Feng X, Pan J - Int J Mol Sci (2015)

Western blot verification of real-time PCR array results. Western blot analysis of cells following a 24 h treatment with RO3280 at 50 or 100 nM compared with DMSO control mock treatment. The upregulation of DCC and CDKN1A and down regulation of BTK and SOCS2 in RO3280 treated cells. Protein lysates from treated cells were tested for expression levels by Western blot analysis.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4307303&req=5

ijms-16-01266-f007: Western blot verification of real-time PCR array results. Western blot analysis of cells following a 24 h treatment with RO3280 at 50 or 100 nM compared with DMSO control mock treatment. The upregulation of DCC and CDKN1A and down regulation of BTK and SOCS2 in RO3280 treated cells. Protein lysates from treated cells were tested for expression levels by Western blot analysis.
Mentions: In order to identify apoptosis and/or programmed cell death molecules implicated in the effects of RO3280, we used the SABioscience Human Apoptosis PCR Array PAHS-3012. With this real-time PCR array we analyzed and clustered the expression of 370 genes associated with apoptosis in DMSO or RO3280 treated cells (Figure 6A). The genes most significantly upregulated or downregulated are shown in Figure 6B,C, respectively. Examination of the array data revealed that 32 genes were significantly upregulated and 16 genes were significantly downregulated in RO3280 treatment group compared with DMSO control group (Table 6 and Table 7, respectively). The upregulated genes included protein kinase C zeta, receptor-interacting serine-threonine kinase 3, harakiri BCL2 interacting protein, DCC netrin 1 receptor, and cyclin-dependent kinase inhibitor 1A. The downregulated genes included interleukin 1α, nucleotide-binding oligomerization domain containing 2, caspase-1, apoptosis-related cysteine peptidase, serpin peptidase inhibitor clade B, B-cell CLL/lymphoma 2, and Bruton agammaglobulinemia tyrosine kinase. The RO3280-dependent upregulation of DCC and CDKN1A and downregulation of BTK and SOCS2 was verified by western blot analysis (Figure 7).

Bottom Line: PLK1 mRNA expression was significantly higher in AML samples compared with control samples (82.95 ± 110.28 vs. 6.36 ± 6.35; p < 0.001).RO3280 treatment regulated several apoptosis-associated genes.These results provide insights into the potential use of RO3280 for AML therapy; however, the underlying mechanisms remain to be determined.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou 215003, China. wangnn90s@163.com.

ABSTRACT
Polo-like kinase 1 (PLK1) is highly expressed in many cancers and therefore a biomarker of transformation and potential target for the development of cancer-specific small molecule drugs. RO3280 was recently identified as a novel PLK1 inhibitor; however its therapeutic effects in leukemia treatment are still unknown. We found that the PLK1 protein was highly expressed in leukemia cell lines as well as 73.3% (11/15) of pediatric acute myeloid leukemia (AML) samples. PLK1 mRNA expression was significantly higher in AML samples compared with control samples (82.95 ± 110.28 vs. 6.36 ± 6.35; p < 0.001). Kaplan-Meier survival analysis revealed that shorter survival time correlated with high tumor PLK1 expression (p = 0.002). The 50% inhibitory concentration (IC50) of RO3280 for acute leukemia cells was between 74 and 797 nM. The IC50 of RO3280 in primary acute lymphocytic leukemia (ALL) and AML cells was between 35.49 and 110.76 nM and 52.80 and 147.50 nM, respectively. RO3280 induced apoptosis and cell cycle disorder in leukemia cells. RO3280 treatment regulated several apoptosis-associated genes. The regulation of DCC, CDKN1A, BTK, and SOCS2 was verified by western blot. These results provide insights into the potential use of RO3280 for AML therapy; however, the underlying mechanisms remain to be determined.

Show MeSH
Related in: MedlinePlus