Limits...
HDAC9 promotes glioblastoma growth via TAZ-mediated EGFR pathway activation.

Yang R, Wu Y, Wang M, Sun Z, Zou J, Zhang Y, Cui H - Oncotarget (2015)

Bottom Line: Also, HDAC9 interacted with TAZ, a key downstream effector of Hippo pathway.Knockdown of HDAC9 decreased the expression of TAZ.We found that overexpressed TAZ in HDAC9-knockdown cells abrogated the effects induced by HDAC9 silencing both in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, P.R. China.

ABSTRACT
Histone deacetylase 9 (HDAC9), a member of class II HDACs, regulates a wide variety of normal and abnormal physiological functions. We found that HDAC9 is over-expressed in prognostically poor glioblastoma patients. Knockdown HDAC9 decreased proliferation in vitro and tumor formation in vivo. HDAC9 accelerated cell cycle in part by potentiating the EGFR signaling pathway. Also, HDAC9 interacted with TAZ, a key downstream effector of Hippo pathway. Knockdown of HDAC9 decreased the expression of TAZ. We found that overexpressed TAZ in HDAC9-knockdown cells abrogated the effects induced by HDAC9 silencing both in vitro and in vivo. We demonstrated that HDAC9 promotes tumor formation of glioblastoma via TAZ-mediated EGFR pathway activation, and provide the evidence for promising target for the treatment of glioblastoma.

No MeSH data available.


Related in: MedlinePlus

Knockdown of HDAC9 inhibits GBM cell growth and proliferation(A) Western blot assay was used to characterize the expression of HDAC9 in HDAC9-knockdown U87 cells. (B) The effect of HDAC9 on the proliferation of U87 cells. (C) The effect of HDAC9 on the viability of U87 cells. (D) Western blot assay was used to characterize the expression of HDAC9 in HDAC9-knockdown LN229 cells. (E) The effect of HDAC9 on the proliferation of LN229 cells. (F) The effect of HDAC9 on the viability of LN229 cells. (G) Western blot assay was used to characterize the expression of HDAC9 in HDAC9-knockdown T0807 cells. (H) The effect of HDAC9 on the proliferation of T0807 cells. (I) The effect of HDAC9 on the viability of T0807 cells. (J, K) Image and quantification of U87 and LN229 cells positive for Brdu staining. All data are shown as the mean ± SD, *p < 0.05, **p < 0.01. All p values are based on analysis control versus treatment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4480706&req=5

Figure 2: Knockdown of HDAC9 inhibits GBM cell growth and proliferation(A) Western blot assay was used to characterize the expression of HDAC9 in HDAC9-knockdown U87 cells. (B) The effect of HDAC9 on the proliferation of U87 cells. (C) The effect of HDAC9 on the viability of U87 cells. (D) Western blot assay was used to characterize the expression of HDAC9 in HDAC9-knockdown LN229 cells. (E) The effect of HDAC9 on the proliferation of LN229 cells. (F) The effect of HDAC9 on the viability of LN229 cells. (G) Western blot assay was used to characterize the expression of HDAC9 in HDAC9-knockdown T0807 cells. (H) The effect of HDAC9 on the proliferation of T0807 cells. (I) The effect of HDAC9 on the viability of T0807 cells. (J, K) Image and quantification of U87 and LN229 cells positive for Brdu staining. All data are shown as the mean ± SD, *p < 0.05, **p < 0.01. All p values are based on analysis control versus treatment.

Mentions: To address the importance of HDAC9 in cell growth and proliferation, we utilized the human glioblastoma cell lines U-87 MG (U87) and LN229, as well as primary cells obtained from GBM patients. Cells were infected with Lentivirus carrying small hairpin RNA (shRNA) constructing against HDAC9 (shHDAC9) or a shGFP control and were subsequently selected by puromycin. Western blot analysis showed that HDAC9 was significantly down-regulated after knockdown by shRNA in different GBM cells (Figure 2A, 2D, 2G). Next, we investigated the proliferation kinetics of GBM cells by using cell growth curve and MTT assay. The growth curve revealed that knockdown of HDAC9 in GBM cells resulted in a significant growth inhibition (Figure 2B and 2H). Furthermore, MTT assay confirmed that down-regulation of HDAC9 induced a significant decrease in cell viability (Figure 2C, 2F and 2I). Above data were confirmed by BrdU incorporation in the U87 and LN229 cell lines, where the HDAC9-knockdown cells showed over a 40% reduction in DNA synthesis compared to control cells in the two cell lines (Figure 2J and 2K). These results demonstrated that HDAC9 was essential for proliferation of GBM cells.


HDAC9 promotes glioblastoma growth via TAZ-mediated EGFR pathway activation.

Yang R, Wu Y, Wang M, Sun Z, Zou J, Zhang Y, Cui H - Oncotarget (2015)

Knockdown of HDAC9 inhibits GBM cell growth and proliferation(A) Western blot assay was used to characterize the expression of HDAC9 in HDAC9-knockdown U87 cells. (B) The effect of HDAC9 on the proliferation of U87 cells. (C) The effect of HDAC9 on the viability of U87 cells. (D) Western blot assay was used to characterize the expression of HDAC9 in HDAC9-knockdown LN229 cells. (E) The effect of HDAC9 on the proliferation of LN229 cells. (F) The effect of HDAC9 on the viability of LN229 cells. (G) Western blot assay was used to characterize the expression of HDAC9 in HDAC9-knockdown T0807 cells. (H) The effect of HDAC9 on the proliferation of T0807 cells. (I) The effect of HDAC9 on the viability of T0807 cells. (J, K) Image and quantification of U87 and LN229 cells positive for Brdu staining. All data are shown as the mean ± SD, *p < 0.05, **p < 0.01. All p values are based on analysis control versus treatment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4480706&req=5

Figure 2: Knockdown of HDAC9 inhibits GBM cell growth and proliferation(A) Western blot assay was used to characterize the expression of HDAC9 in HDAC9-knockdown U87 cells. (B) The effect of HDAC9 on the proliferation of U87 cells. (C) The effect of HDAC9 on the viability of U87 cells. (D) Western blot assay was used to characterize the expression of HDAC9 in HDAC9-knockdown LN229 cells. (E) The effect of HDAC9 on the proliferation of LN229 cells. (F) The effect of HDAC9 on the viability of LN229 cells. (G) Western blot assay was used to characterize the expression of HDAC9 in HDAC9-knockdown T0807 cells. (H) The effect of HDAC9 on the proliferation of T0807 cells. (I) The effect of HDAC9 on the viability of T0807 cells. (J, K) Image and quantification of U87 and LN229 cells positive for Brdu staining. All data are shown as the mean ± SD, *p < 0.05, **p < 0.01. All p values are based on analysis control versus treatment.
Mentions: To address the importance of HDAC9 in cell growth and proliferation, we utilized the human glioblastoma cell lines U-87 MG (U87) and LN229, as well as primary cells obtained from GBM patients. Cells were infected with Lentivirus carrying small hairpin RNA (shRNA) constructing against HDAC9 (shHDAC9) or a shGFP control and were subsequently selected by puromycin. Western blot analysis showed that HDAC9 was significantly down-regulated after knockdown by shRNA in different GBM cells (Figure 2A, 2D, 2G). Next, we investigated the proliferation kinetics of GBM cells by using cell growth curve and MTT assay. The growth curve revealed that knockdown of HDAC9 in GBM cells resulted in a significant growth inhibition (Figure 2B and 2H). Furthermore, MTT assay confirmed that down-regulation of HDAC9 induced a significant decrease in cell viability (Figure 2C, 2F and 2I). Above data were confirmed by BrdU incorporation in the U87 and LN229 cell lines, where the HDAC9-knockdown cells showed over a 40% reduction in DNA synthesis compared to control cells in the two cell lines (Figure 2J and 2K). These results demonstrated that HDAC9 was essential for proliferation of GBM cells.

Bottom Line: Also, HDAC9 interacted with TAZ, a key downstream effector of Hippo pathway.Knockdown of HDAC9 decreased the expression of TAZ.We found that overexpressed TAZ in HDAC9-knockdown cells abrogated the effects induced by HDAC9 silencing both in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, P.R. China.

ABSTRACT
Histone deacetylase 9 (HDAC9), a member of class II HDACs, regulates a wide variety of normal and abnormal physiological functions. We found that HDAC9 is over-expressed in prognostically poor glioblastoma patients. Knockdown HDAC9 decreased proliferation in vitro and tumor formation in vivo. HDAC9 accelerated cell cycle in part by potentiating the EGFR signaling pathway. Also, HDAC9 interacted with TAZ, a key downstream effector of Hippo pathway. Knockdown of HDAC9 decreased the expression of TAZ. We found that overexpressed TAZ in HDAC9-knockdown cells abrogated the effects induced by HDAC9 silencing both in vitro and in vivo. We demonstrated that HDAC9 promotes tumor formation of glioblastoma via TAZ-mediated EGFR pathway activation, and provide the evidence for promising target for the treatment of glioblastoma.

No MeSH data available.


Related in: MedlinePlus