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Knockdown of a novel lincRNA AATBC suppresses proliferation and induces apoptosis in bladder cancer.

Zhao F, Lin T, He W, Han J, Zhu D, Hu K, Li W, Zheng Z, Huang J, Xie W - Oncotarget (2015)

Bottom Line: We also found that inhibition of AATBC resulted in cell proliferation arrest through G1 cell cycle mediated by cyclin D1, CDK4, p18 and phosphorylated Rb.Furthermore, JNK inhibitor SP600125 could attenuate the apoptotic effect achieved by AATBC knockdown, confirming the involvement of JNK signaling in the induced apoptosis.Moreover, mouse xenograft model revealed that knockdown of AATBC led to suppress tumorigenesis in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

ABSTRACT
Long intergenic noncoding RNAs (lincRNAs) play important roles in regulating various biological processes in cancer, including proliferation and apoptosis. However, the roles of lincRNAs in bladder cancer remain elusive. In this study, we identified a novel lincRNA, which we termed AATBC. We found that AATBC was overexpressed in bladder cancer patient tissues and positively correlated with tumor grade and pT stage. We also found that inhibition of AATBC resulted in cell proliferation arrest through G1 cell cycle mediated by cyclin D1, CDK4, p18 and phosphorylated Rb. In addition, inhibition of AATBC induced cell apoptosis through the intrinsic apoptosis signaling pathway, as evidenced by the activation of caspase-9 and caspase-3. The investigation for the signaling pathway revealed that the apoptosis following AATBC knockdown was mediated by activation of phosphorylated JNK and suppression of NRF2. Furthermore, JNK inhibitor SP600125 could attenuate the apoptotic effect achieved by AATBC knockdown, confirming the involvement of JNK signaling in the induced apoptosis. Moreover, mouse xenograft model revealed that knockdown of AATBC led to suppress tumorigenesis in vivo. Taken together, our study indicated that AATBC might play a critical role in pro-proliferation and anti-apoptosis in bladder cancer by regulating cell cycle, intrinsic apoptosis signaling, JNK signaling and NRF2. AATBC could be a potential therapeutic target and molecular biomarker for bladder cancer.

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AATBC knockdown attenuated bladder cancer cell proliferationA. AATBC expression level was evaluated by quantitative RT-PCR in UM-UC-3 and EJ bladder cancer cell lines after transfection with siRNAs and negative controls. Means ± SD were shown (n = 3). *P<0.05 (vs. control). B. AATBC knockdown attenuated bladder cancer cell lines UM-UC-3 and EJ proliferation as determined by CellTiter 96 AQueous One Solution Cell Proliferation Assay. Means ± SD were shown (n = 3). *P<0.05 (vs. control). C. Colony formation by AATBC knockdown bladder cancer cells. Histological analysis of the rate of colony formation in control and AATBC knockdown groups. Means ± SD were shown (n = 3). *P<0.05 (vs. control). D. Analysis of the percent of EdU positive cells in negative control and AATBC knockdown groups in UM-UC-3 and EJ cells. Means ± SD were shown (n = 3). *P<0.05 (vs. control). AATBC knockdown led to decrease in S phase in both UM-UC-3 and EJ cells. Blue color represented the nucleus and red color indicated S phase cells (EdU positive).
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Figure 2: AATBC knockdown attenuated bladder cancer cell proliferationA. AATBC expression level was evaluated by quantitative RT-PCR in UM-UC-3 and EJ bladder cancer cell lines after transfection with siRNAs and negative controls. Means ± SD were shown (n = 3). *P<0.05 (vs. control). B. AATBC knockdown attenuated bladder cancer cell lines UM-UC-3 and EJ proliferation as determined by CellTiter 96 AQueous One Solution Cell Proliferation Assay. Means ± SD were shown (n = 3). *P<0.05 (vs. control). C. Colony formation by AATBC knockdown bladder cancer cells. Histological analysis of the rate of colony formation in control and AATBC knockdown groups. Means ± SD were shown (n = 3). *P<0.05 (vs. control). D. Analysis of the percent of EdU positive cells in negative control and AATBC knockdown groups in UM-UC-3 and EJ cells. Means ± SD were shown (n = 3). *P<0.05 (vs. control). AATBC knockdown led to decrease in S phase in both UM-UC-3 and EJ cells. Blue color represented the nucleus and red color indicated S phase cells (EdU positive).

Mentions: We knocked down AATBC in UM-UC-3 and EJ bladder cancer cells using small interfering RNAs (siRNAs). The knockdown efficiency in UM-UC-3 cells was si#1 64.3% ± 3.0% and si#2 81.0% ± 3.6%. In EJ cells, the knockdown efficiency was si#1 61.7% ± 3.5% and si#2 77.7% ± 4.9% (Fig. 2A). In order to investigate the effect of AATBC on cell growth of bladder cancer cell lines in vitro, CellTiter 96 AQueous One Solution Cell Proliferation assay, panel clonogenic assay, EdU incorporation assay and flow cytometry were employed. These results show that AATBC depletion led to an obvious inhibitory effect on the growth of bladder cancer cells.


Knockdown of a novel lincRNA AATBC suppresses proliferation and induces apoptosis in bladder cancer.

Zhao F, Lin T, He W, Han J, Zhu D, Hu K, Li W, Zheng Z, Huang J, Xie W - Oncotarget (2015)

AATBC knockdown attenuated bladder cancer cell proliferationA. AATBC expression level was evaluated by quantitative RT-PCR in UM-UC-3 and EJ bladder cancer cell lines after transfection with siRNAs and negative controls. Means ± SD were shown (n = 3). *P<0.05 (vs. control). B. AATBC knockdown attenuated bladder cancer cell lines UM-UC-3 and EJ proliferation as determined by CellTiter 96 AQueous One Solution Cell Proliferation Assay. Means ± SD were shown (n = 3). *P<0.05 (vs. control). C. Colony formation by AATBC knockdown bladder cancer cells. Histological analysis of the rate of colony formation in control and AATBC knockdown groups. Means ± SD were shown (n = 3). *P<0.05 (vs. control). D. Analysis of the percent of EdU positive cells in negative control and AATBC knockdown groups in UM-UC-3 and EJ cells. Means ± SD were shown (n = 3). *P<0.05 (vs. control). AATBC knockdown led to decrease in S phase in both UM-UC-3 and EJ cells. Blue color represented the nucleus and red color indicated S phase cells (EdU positive).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4359217&req=5

Figure 2: AATBC knockdown attenuated bladder cancer cell proliferationA. AATBC expression level was evaluated by quantitative RT-PCR in UM-UC-3 and EJ bladder cancer cell lines after transfection with siRNAs and negative controls. Means ± SD were shown (n = 3). *P<0.05 (vs. control). B. AATBC knockdown attenuated bladder cancer cell lines UM-UC-3 and EJ proliferation as determined by CellTiter 96 AQueous One Solution Cell Proliferation Assay. Means ± SD were shown (n = 3). *P<0.05 (vs. control). C. Colony formation by AATBC knockdown bladder cancer cells. Histological analysis of the rate of colony formation in control and AATBC knockdown groups. Means ± SD were shown (n = 3). *P<0.05 (vs. control). D. Analysis of the percent of EdU positive cells in negative control and AATBC knockdown groups in UM-UC-3 and EJ cells. Means ± SD were shown (n = 3). *P<0.05 (vs. control). AATBC knockdown led to decrease in S phase in both UM-UC-3 and EJ cells. Blue color represented the nucleus and red color indicated S phase cells (EdU positive).
Mentions: We knocked down AATBC in UM-UC-3 and EJ bladder cancer cells using small interfering RNAs (siRNAs). The knockdown efficiency in UM-UC-3 cells was si#1 64.3% ± 3.0% and si#2 81.0% ± 3.6%. In EJ cells, the knockdown efficiency was si#1 61.7% ± 3.5% and si#2 77.7% ± 4.9% (Fig. 2A). In order to investigate the effect of AATBC on cell growth of bladder cancer cell lines in vitro, CellTiter 96 AQueous One Solution Cell Proliferation assay, panel clonogenic assay, EdU incorporation assay and flow cytometry were employed. These results show that AATBC depletion led to an obvious inhibitory effect on the growth of bladder cancer cells.

Bottom Line: We also found that inhibition of AATBC resulted in cell proliferation arrest through G1 cell cycle mediated by cyclin D1, CDK4, p18 and phosphorylated Rb.Furthermore, JNK inhibitor SP600125 could attenuate the apoptotic effect achieved by AATBC knockdown, confirming the involvement of JNK signaling in the induced apoptosis.Moreover, mouse xenograft model revealed that knockdown of AATBC led to suppress tumorigenesis in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

ABSTRACT
Long intergenic noncoding RNAs (lincRNAs) play important roles in regulating various biological processes in cancer, including proliferation and apoptosis. However, the roles of lincRNAs in bladder cancer remain elusive. In this study, we identified a novel lincRNA, which we termed AATBC. We found that AATBC was overexpressed in bladder cancer patient tissues and positively correlated with tumor grade and pT stage. We also found that inhibition of AATBC resulted in cell proliferation arrest through G1 cell cycle mediated by cyclin D1, CDK4, p18 and phosphorylated Rb. In addition, inhibition of AATBC induced cell apoptosis through the intrinsic apoptosis signaling pathway, as evidenced by the activation of caspase-9 and caspase-3. The investigation for the signaling pathway revealed that the apoptosis following AATBC knockdown was mediated by activation of phosphorylated JNK and suppression of NRF2. Furthermore, JNK inhibitor SP600125 could attenuate the apoptotic effect achieved by AATBC knockdown, confirming the involvement of JNK signaling in the induced apoptosis. Moreover, mouse xenograft model revealed that knockdown of AATBC led to suppress tumorigenesis in vivo. Taken together, our study indicated that AATBC might play a critical role in pro-proliferation and anti-apoptosis in bladder cancer by regulating cell cycle, intrinsic apoptosis signaling, JNK signaling and NRF2. AATBC could be a potential therapeutic target and molecular biomarker for bladder cancer.

Show MeSH
Related in: MedlinePlus