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The miR-193a-3p-regulated ING5 gene activates the DNA damage response pathway and inhibits multi-chemoresistance in bladder cancer.

Li Y, Deng H, Lv L, Zhang C, Qian L, Xiao J, Zhao W, Liu Q, Zhang D, Wang Y, Yan J, Zhang H, He Y, Zhu J - Oncotarget (2015)

Bottom Line: We have previously shown that the bladder cancer (BCa) cell line 5637 is significantly more sensitive to the cytoxicity of five chemotherapeutic agents than H-bc cells.We also showed that ING5 gene expression is inhibited by miR-193a-3p and is instrumental in miR-193a-3p's role in activating BCa chemoresistance.Our results demonstrate both the role and mechanism of inhibition of BCa chemoresistance by ING5.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, School of Life Science, Anhui Medical University, Hefei, Anhui, China.

ABSTRACT
As the major barrier to curative cancer chemotherapy, chemoresistance presents a formidable challenge to both cancer researchers and clinicians. We have previously shown that the bladder cancer (BCa) cell line 5637 is significantly more sensitive to the cytoxicity of five chemotherapeutic agents than H-bc cells. Using an RNA-seq-based omic analysis and validation at both the mRNA and protein levels, we found that the inhibitor of growth 5 (ING5) gene was upregulated in 5637 cells compared with H-bc cells, indicating that it has an inhibitory role in BCa chemoresistance. siRNA-mediated inhibition of ING5 increased the chemoresistance and inhibited the DNA damage response pathway in 5637 cells. Conversely, forced expression of EGFP-ING5 decreased the chemoresistance of and activated the DNA damage response pathway in H-bc cells. We also showed that ING5 gene expression is inhibited by miR-193a-3p and is instrumental in miR-193a-3p's role in activating BCa chemoresistance. Our results demonstrate both the role and mechanism of inhibition of BCa chemoresistance by ING5.

No MeSH data available.


Related in: MedlinePlus

ING5 expression is negatively correlated with chemoresistance in BCa cellsA, ING5 level in 5637 versus H-bc cells by RNA-seq, qRT-PCR, and Western blot analyses. B, Graph of RNA levels by RNA-seq and qRT-PCR. C, Western blot of ING5 in 5637 and H-bc cells. D, The ING5 mRNA (by qRT-PCR) and protein (by Western analysis) levels in the each of three ING5 siRNA (si-ING5-1, -2 and -3)-transfected versus the NC-transfected 5637 cells. E, The relative cell survival in the 5637 cells transfected with ING5 siRNAs vs. NC siRNA, assayed 72 hours after treatment with IC50-dosed drug (Pa, Ad, Ci, Pi or EH). F, The ING5 protein level in the EGFP-ING5-transfected versus the GFP (control)-transfected H-bc cells. G, The relative cell survival of the H-bc cells transfected with EGFP-ING5 versus GFP expression vector, assayed 72 hours after treatment with IC50-dosed drug. (*P<0.05; **P<0.01).
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Figure 1: ING5 expression is negatively correlated with chemoresistance in BCa cellsA, ING5 level in 5637 versus H-bc cells by RNA-seq, qRT-PCR, and Western blot analyses. B, Graph of RNA levels by RNA-seq and qRT-PCR. C, Western blot of ING5 in 5637 and H-bc cells. D, The ING5 mRNA (by qRT-PCR) and protein (by Western analysis) levels in the each of three ING5 siRNA (si-ING5-1, -2 and -3)-transfected versus the NC-transfected 5637 cells. E, The relative cell survival in the 5637 cells transfected with ING5 siRNAs vs. NC siRNA, assayed 72 hours after treatment with IC50-dosed drug (Pa, Ad, Ci, Pi or EH). F, The ING5 protein level in the EGFP-ING5-transfected versus the GFP (control)-transfected H-bc cells. G, The relative cell survival of the H-bc cells transfected with EGFP-ING5 versus GFP expression vector, assayed 72 hours after treatment with IC50-dosed drug. (*P<0.05; **P<0.01).

Mentions: Among 9051 differentially expressed genes between a multi-chemosensitive (5637) and a resistant BCa cell line (H-bc) that were revealed by an RNA-seq-based omic study (data not shown), ING5 was among the top 10% most differentially expressed (Fig. 1A). The BCa chemoresistance-associated expression of the ING5 gene was confirmed by qRT-PCR analysis at the steady state mRNA level and a Western blot analysis at the protein level. The results showed that the ING5 protein (1.00:0.61, Fig. 1A and C) and mRNA expression levels were higher (RNA-seq-based omic: 1.00:0.28, and qRT-PCR analysis: 1.00:0.72, Fig. 1A and B) in 5637 cells than in H-bc cells.


The miR-193a-3p-regulated ING5 gene activates the DNA damage response pathway and inhibits multi-chemoresistance in bladder cancer.

Li Y, Deng H, Lv L, Zhang C, Qian L, Xiao J, Zhao W, Liu Q, Zhang D, Wang Y, Yan J, Zhang H, He Y, Zhu J - Oncotarget (2015)

ING5 expression is negatively correlated with chemoresistance in BCa cellsA, ING5 level in 5637 versus H-bc cells by RNA-seq, qRT-PCR, and Western blot analyses. B, Graph of RNA levels by RNA-seq and qRT-PCR. C, Western blot of ING5 in 5637 and H-bc cells. D, The ING5 mRNA (by qRT-PCR) and protein (by Western analysis) levels in the each of three ING5 siRNA (si-ING5-1, -2 and -3)-transfected versus the NC-transfected 5637 cells. E, The relative cell survival in the 5637 cells transfected with ING5 siRNAs vs. NC siRNA, assayed 72 hours after treatment with IC50-dosed drug (Pa, Ad, Ci, Pi or EH). F, The ING5 protein level in the EGFP-ING5-transfected versus the GFP (control)-transfected H-bc cells. G, The relative cell survival of the H-bc cells transfected with EGFP-ING5 versus GFP expression vector, assayed 72 hours after treatment with IC50-dosed drug. (*P<0.05; **P<0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4496349&req=5

Figure 1: ING5 expression is negatively correlated with chemoresistance in BCa cellsA, ING5 level in 5637 versus H-bc cells by RNA-seq, qRT-PCR, and Western blot analyses. B, Graph of RNA levels by RNA-seq and qRT-PCR. C, Western blot of ING5 in 5637 and H-bc cells. D, The ING5 mRNA (by qRT-PCR) and protein (by Western analysis) levels in the each of three ING5 siRNA (si-ING5-1, -2 and -3)-transfected versus the NC-transfected 5637 cells. E, The relative cell survival in the 5637 cells transfected with ING5 siRNAs vs. NC siRNA, assayed 72 hours after treatment with IC50-dosed drug (Pa, Ad, Ci, Pi or EH). F, The ING5 protein level in the EGFP-ING5-transfected versus the GFP (control)-transfected H-bc cells. G, The relative cell survival of the H-bc cells transfected with EGFP-ING5 versus GFP expression vector, assayed 72 hours after treatment with IC50-dosed drug. (*P<0.05; **P<0.01).
Mentions: Among 9051 differentially expressed genes between a multi-chemosensitive (5637) and a resistant BCa cell line (H-bc) that were revealed by an RNA-seq-based omic study (data not shown), ING5 was among the top 10% most differentially expressed (Fig. 1A). The BCa chemoresistance-associated expression of the ING5 gene was confirmed by qRT-PCR analysis at the steady state mRNA level and a Western blot analysis at the protein level. The results showed that the ING5 protein (1.00:0.61, Fig. 1A and C) and mRNA expression levels were higher (RNA-seq-based omic: 1.00:0.28, and qRT-PCR analysis: 1.00:0.72, Fig. 1A and B) in 5637 cells than in H-bc cells.

Bottom Line: We have previously shown that the bladder cancer (BCa) cell line 5637 is significantly more sensitive to the cytoxicity of five chemotherapeutic agents than H-bc cells.We also showed that ING5 gene expression is inhibited by miR-193a-3p and is instrumental in miR-193a-3p's role in activating BCa chemoresistance.Our results demonstrate both the role and mechanism of inhibition of BCa chemoresistance by ING5.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, School of Life Science, Anhui Medical University, Hefei, Anhui, China.

ABSTRACT
As the major barrier to curative cancer chemotherapy, chemoresistance presents a formidable challenge to both cancer researchers and clinicians. We have previously shown that the bladder cancer (BCa) cell line 5637 is significantly more sensitive to the cytoxicity of five chemotherapeutic agents than H-bc cells. Using an RNA-seq-based omic analysis and validation at both the mRNA and protein levels, we found that the inhibitor of growth 5 (ING5) gene was upregulated in 5637 cells compared with H-bc cells, indicating that it has an inhibitory role in BCa chemoresistance. siRNA-mediated inhibition of ING5 increased the chemoresistance and inhibited the DNA damage response pathway in 5637 cells. Conversely, forced expression of EGFP-ING5 decreased the chemoresistance of and activated the DNA damage response pathway in H-bc cells. We also showed that ING5 gene expression is inhibited by miR-193a-3p and is instrumental in miR-193a-3p's role in activating BCa chemoresistance. Our results demonstrate both the role and mechanism of inhibition of BCa chemoresistance by ING5.

No MeSH data available.


Related in: MedlinePlus