Limits...
Tubulin Beta3 Serves as a Target of HDAC3 and Mediates Resistance to Microtubule-Targeting Drugs.

Kim Y, Kim H, Jeoung D - Mol. Cells (2015)

Bottom Line: The down-regulation of HDAC6 decreased the expression of MDR1 and tubulin β3, but did not affect HDAC3 expression.The down-regulation of tubulin β3 did not affect the expression of HDAC6 or MDR1.Our results showed that tubulin β3 serves as a downstream target of HDAC3 and mediates resistance to microtubule-targeting drugs.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, College of Natural Sciences, Kangwon National University, Chunchon 200-701, Korea.

ABSTRACT
We investigated the role of HDAC3 in anti-cancer drug-resistance. The expression of HDAC3 was decreased in cancer cell lines resistant to anti-cancer drugs such as celastrol and taxol. HDAC3 conferred sensitivity to these anti-cancer drugs. HDAC3 activity was necessary for conferring sensitivity to these anti-cancer drugs. The down-regulation of HDAC3 increased the expression of MDR1 and conferred resistance to anti-cancer drugs. The expression of tubulin β3 was increased in drug-resistant cancer cell lines. ChIP assays showed the binding of HDAC3 to the promoter sequences of tubulin β3 and HDAC6. HDAC6 showed an interaction with tubulin β3. HDAC3 had a negative regulatory role in the expression of tubulin β3 and HDAC6. The down-regulation of HDAC6 decreased the expression of MDR1 and tubulin β3, but did not affect HDAC3 expression. The down-regulation of HDAC6 conferred sensitivity to taxol. The down-regulation of tubulin β3 did not affect the expression of HDAC6 or MDR1. The down-regulation of tubulin β3 conferred sensitivity to anti-cancer drugs. Our results showed that tubulin β3 serves as a downstream target of HDAC3 and mediates resistance to microtubule-targeting drugs. Thus, the HDAC3-HDAC6-Tubulin β axis can be employed for the development of anti-cancer drugs.

No MeSH data available.


Related in: MedlinePlus

Wild type, but not mutant HDAC3, confers sensitivity to microtubule-disrupting drugs in cancer cell lines that stably express antisense-HDAC3. Cell lysates from the indicated cell line were subjected to Western blot (A, B). (C) SNU387 cells that stably express antsense-HDAC3 (SNU387-As-HDAC3) were transiently transfected with control vector, HDAC3-Myc/His or HDAC3S424A-Myc/His. At 24 h after transfection, cells were treated with or without celastrol (1 μM), taxol (1 μM) or vinblastine (100 nM) for 24 h, followed by Western blot analysis. (D) The same as C except that Malme3M-As-HDAC3 cell line was employed.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4546942&req=5

f3-molce-38-8-705: Wild type, but not mutant HDAC3, confers sensitivity to microtubule-disrupting drugs in cancer cell lines that stably express antisense-HDAC3. Cell lysates from the indicated cell line were subjected to Western blot (A, B). (C) SNU387 cells that stably express antsense-HDAC3 (SNU387-As-HDAC3) were transiently transfected with control vector, HDAC3-Myc/His or HDAC3S424A-Myc/His. At 24 h after transfection, cells were treated with or without celastrol (1 μM), taxol (1 μM) or vinblastine (100 nM) for 24 h, followed by Western blot analysis. (D) The same as C except that Malme3M-As-HDAC3 cell line was employed.

Mentions: Because the down-regulation of HDAC 3was correlated with resistance to taxol, we hypothesized that over-expression of HDAC3 confers sensitivity to microtubule-targeting drugs. In this study, we employed catalytically inactive mutant HDAC3S424A to determine whether HDAC3 activity is necessary to confer sensitivity to microtubule-targeting drugs. Ser424 is a non-conserved residue among class I HDACs. Ser 424 is the the protein kinase CK2 phosphoacceptor site in HDAC3 (Zhang et al., 2005). HDAC3Ser424 lacks histone deacetylase activity (Zhang et al., 2005). Wild type HDAC3, but not HDAC3S424A, enhanced the sensitivity of HepG2, Hep3B and WM-266-4 cells (Figs. 2A, 2C, and 2E) to microtubule-targeting drugs such as celsatrol, taxol and vinblastine. The enhanced sensitivity was accompanied by enhanced cleavages of PARP and caspase-3 in these cancer cells (Figs. 2B, 2D, and 2F). These results suggested that HDAC3 activity is necessary for conferring sensitivity to microtubule-targeting drugs. We established cancer cell lines that stably express anti-sense HDAC3 (SNU387-As-HDAC3, Mame3MAs-HDAC3) to further confirm the role of HDAC3. SNU387-As-HDAC3 cells and Malme3M-As-HDAC3 cells showed higher resistance to microtubule-targeting drugs than the respective controls (Table 1). They also showed increased expression of MDR1 (Figs. 3A and 3B). Wild type HDAC3, but not HDAC3Ser424A, enhanced cleavages of PARP and caspase-3 in SNU387-As-HDAC3 and Malme3M-As-HDAC3 in response to microtubule-targeting drugs (Figs. 3C and 3D). Wild type HDAC3, but not HDAC3S424A, enhanced the sensitivity of SNU387-As-HDAC3 cells and Malme3M-As-HDAC3 cells to microtubule-targeting drugs (Table 1). This result confirmed that resistance to microtubule-targeting drugs results from the down-regulation of HDAC3. Taken together, these results suggested that HDAC3 regulates response to microtubule-targeting drugs.


Tubulin Beta3 Serves as a Target of HDAC3 and Mediates Resistance to Microtubule-Targeting Drugs.

Kim Y, Kim H, Jeoung D - Mol. Cells (2015)

Wild type, but not mutant HDAC3, confers sensitivity to microtubule-disrupting drugs in cancer cell lines that stably express antisense-HDAC3. Cell lysates from the indicated cell line were subjected to Western blot (A, B). (C) SNU387 cells that stably express antsense-HDAC3 (SNU387-As-HDAC3) were transiently transfected with control vector, HDAC3-Myc/His or HDAC3S424A-Myc/His. At 24 h after transfection, cells were treated with or without celastrol (1 μM), taxol (1 μM) or vinblastine (100 nM) for 24 h, followed by Western blot analysis. (D) The same as C except that Malme3M-As-HDAC3 cell line was employed.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4546942&req=5

f3-molce-38-8-705: Wild type, but not mutant HDAC3, confers sensitivity to microtubule-disrupting drugs in cancer cell lines that stably express antisense-HDAC3. Cell lysates from the indicated cell line were subjected to Western blot (A, B). (C) SNU387 cells that stably express antsense-HDAC3 (SNU387-As-HDAC3) were transiently transfected with control vector, HDAC3-Myc/His or HDAC3S424A-Myc/His. At 24 h after transfection, cells were treated with or without celastrol (1 μM), taxol (1 μM) or vinblastine (100 nM) for 24 h, followed by Western blot analysis. (D) The same as C except that Malme3M-As-HDAC3 cell line was employed.
Mentions: Because the down-regulation of HDAC 3was correlated with resistance to taxol, we hypothesized that over-expression of HDAC3 confers sensitivity to microtubule-targeting drugs. In this study, we employed catalytically inactive mutant HDAC3S424A to determine whether HDAC3 activity is necessary to confer sensitivity to microtubule-targeting drugs. Ser424 is a non-conserved residue among class I HDACs. Ser 424 is the the protein kinase CK2 phosphoacceptor site in HDAC3 (Zhang et al., 2005). HDAC3Ser424 lacks histone deacetylase activity (Zhang et al., 2005). Wild type HDAC3, but not HDAC3S424A, enhanced the sensitivity of HepG2, Hep3B and WM-266-4 cells (Figs. 2A, 2C, and 2E) to microtubule-targeting drugs such as celsatrol, taxol and vinblastine. The enhanced sensitivity was accompanied by enhanced cleavages of PARP and caspase-3 in these cancer cells (Figs. 2B, 2D, and 2F). These results suggested that HDAC3 activity is necessary for conferring sensitivity to microtubule-targeting drugs. We established cancer cell lines that stably express anti-sense HDAC3 (SNU387-As-HDAC3, Mame3MAs-HDAC3) to further confirm the role of HDAC3. SNU387-As-HDAC3 cells and Malme3M-As-HDAC3 cells showed higher resistance to microtubule-targeting drugs than the respective controls (Table 1). They also showed increased expression of MDR1 (Figs. 3A and 3B). Wild type HDAC3, but not HDAC3Ser424A, enhanced cleavages of PARP and caspase-3 in SNU387-As-HDAC3 and Malme3M-As-HDAC3 in response to microtubule-targeting drugs (Figs. 3C and 3D). Wild type HDAC3, but not HDAC3S424A, enhanced the sensitivity of SNU387-As-HDAC3 cells and Malme3M-As-HDAC3 cells to microtubule-targeting drugs (Table 1). This result confirmed that resistance to microtubule-targeting drugs results from the down-regulation of HDAC3. Taken together, these results suggested that HDAC3 regulates response to microtubule-targeting drugs.

Bottom Line: The down-regulation of HDAC6 decreased the expression of MDR1 and tubulin β3, but did not affect HDAC3 expression.The down-regulation of tubulin β3 did not affect the expression of HDAC6 or MDR1.Our results showed that tubulin β3 serves as a downstream target of HDAC3 and mediates resistance to microtubule-targeting drugs.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, College of Natural Sciences, Kangwon National University, Chunchon 200-701, Korea.

ABSTRACT
We investigated the role of HDAC3 in anti-cancer drug-resistance. The expression of HDAC3 was decreased in cancer cell lines resistant to anti-cancer drugs such as celastrol and taxol. HDAC3 conferred sensitivity to these anti-cancer drugs. HDAC3 activity was necessary for conferring sensitivity to these anti-cancer drugs. The down-regulation of HDAC3 increased the expression of MDR1 and conferred resistance to anti-cancer drugs. The expression of tubulin β3 was increased in drug-resistant cancer cell lines. ChIP assays showed the binding of HDAC3 to the promoter sequences of tubulin β3 and HDAC6. HDAC6 showed an interaction with tubulin β3. HDAC3 had a negative regulatory role in the expression of tubulin β3 and HDAC6. The down-regulation of HDAC6 decreased the expression of MDR1 and tubulin β3, but did not affect HDAC3 expression. The down-regulation of HDAC6 conferred sensitivity to taxol. The down-regulation of tubulin β3 did not affect the expression of HDAC6 or MDR1. The down-regulation of tubulin β3 conferred sensitivity to anti-cancer drugs. Our results showed that tubulin β3 serves as a downstream target of HDAC3 and mediates resistance to microtubule-targeting drugs. Thus, the HDAC3-HDAC6-Tubulin β axis can be employed for the development of anti-cancer drugs.

No MeSH data available.


Related in: MedlinePlus