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N‑Myc downstream‑regulated gene 2 suppresses the proliferation of T24 human bladder cancer cells via induction of oncosis.

Huang J, Wu Z, Wang G, Cai Y, Cai M, Li Y - Mol Med Rep (2015)

Bottom Line: The proliferation rate of T24 cells was significantly reduced by NDRG2 expression (P<0.01).In addition, 82.1% of NDRG2‑expressing cells were in S‑phase, compared to 74.4% in the control virus‑infected cells (P<0.05).In conclusion, the results of the present study indicated that NDRG2 expression in mitochondria may arrest bladder cancer cells in S‑phase as well as decrease cell proliferation through inducing oncosis.

View Article: PubMed Central - PubMed

Affiliation: Cancer Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong 524001, P.R. China.

ABSTRACT
Previous studies have reported the antitumor activity of N‑Myc downstream‑regulated gene 2 (NDRG2), a novel p53‑inducible gene, in several types of cancer. The present study aimed to investigate the effects of NDRG2 expression on the proliferation of a human bladder cancer cell line. NDRG2 and control green fluorescent protein (GFP) recombinant adenovirus plasmids were constructed and transfected into a bladder cancer cell line with mutant p53 (T24 cells). NDRG2 expression was analyzed using western blot analysis and immunofluorescence assay (IFA); in addition, the subcellular localization of NDRG2 was detected using a confocal microscope. The proliferation rate of cells was measured using colony formation and MTT assays. Furthermore, the cell cycle of transfected T24 cells was detected by flow cytometry. The results indicated that T24 cells expressed low levels of NDRG2 prior to infection with GFP‑NDRG2 recombinant adenovirus; by contrast, following infection, NDRG2 was primarily overexpressed in mitochondria. The proliferation rate of T24 cells was significantly reduced by NDRG2 expression (P<0.01). In addition, 82.1% of NDRG2‑expressing cells were in S‑phase, compared to 74.4% in the control virus‑infected cells (P<0.05). Furthermore, upregulation of NDRG2 induced an increase in oncosis, rather than apoptosis, in T24 cell. In conclusion, the results of the present study indicated that NDRG2 expression in mitochondria may arrest bladder cancer cells in S‑phase as well as decrease cell proliferation through inducing oncosis. It was therefore proposed that NDRG2 was not only a biomarker, but also a tumor suppressor for bladder cancer.

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Subcellular localization of NDRG2 protein following gene transfection. (A) Subcellular localization of NDRG2 protein in the mitochondria of the NDRG2-transfected cells at 6, 16, 24, 48 and 72 h following transfection. (B) Quantification of NDRG2 protein expression following transfection. Values are presented as the mean ± standard deviation. *P<0.05 and **P<0.01 vs. 6 h. NDRG2, N-Myc dowstream-regulated gene 2; MFI, mean fluorescence intensity.
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f3-mmr-12-04-5730: Subcellular localization of NDRG2 protein following gene transfection. (A) Subcellular localization of NDRG2 protein in the mitochondria of the NDRG2-transfected cells at 6, 16, 24, 48 and 72 h following transfection. (B) Quantification of NDRG2 protein expression following transfection. Values are presented as the mean ± standard deviation. *P<0.05 and **P<0.01 vs. 6 h. NDRG2, N-Myc dowstream-regulated gene 2; MFI, mean fluorescence intensity.

Mentions: In order to evaluate the subcellular localization of NDRG2, cells were stained with organelle-specific dyes and then visualized using a confocal laser fluorescence microscope. As shown in Fig. 3A, following infection with the GFP-NDRG2 virus, NDRG2 was observed to be expressed primarily in the mitochondria and was not co-localized with the golgi apparatus or lysosomes. In addition, the expression of NDRG2 in the mitochondria was observed to be increased significantly from at 24, 48 and 72 h post infection compared with 6 h post infection (P<0.05, 0.05 and 0.01, respectively) (Fig. 3B).


N‑Myc downstream‑regulated gene 2 suppresses the proliferation of T24 human bladder cancer cells via induction of oncosis.

Huang J, Wu Z, Wang G, Cai Y, Cai M, Li Y - Mol Med Rep (2015)

Subcellular localization of NDRG2 protein following gene transfection. (A) Subcellular localization of NDRG2 protein in the mitochondria of the NDRG2-transfected cells at 6, 16, 24, 48 and 72 h following transfection. (B) Quantification of NDRG2 protein expression following transfection. Values are presented as the mean ± standard deviation. *P<0.05 and **P<0.01 vs. 6 h. NDRG2, N-Myc dowstream-regulated gene 2; MFI, mean fluorescence intensity.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581762&req=5

f3-mmr-12-04-5730: Subcellular localization of NDRG2 protein following gene transfection. (A) Subcellular localization of NDRG2 protein in the mitochondria of the NDRG2-transfected cells at 6, 16, 24, 48 and 72 h following transfection. (B) Quantification of NDRG2 protein expression following transfection. Values are presented as the mean ± standard deviation. *P<0.05 and **P<0.01 vs. 6 h. NDRG2, N-Myc dowstream-regulated gene 2; MFI, mean fluorescence intensity.
Mentions: In order to evaluate the subcellular localization of NDRG2, cells were stained with organelle-specific dyes and then visualized using a confocal laser fluorescence microscope. As shown in Fig. 3A, following infection with the GFP-NDRG2 virus, NDRG2 was observed to be expressed primarily in the mitochondria and was not co-localized with the golgi apparatus or lysosomes. In addition, the expression of NDRG2 in the mitochondria was observed to be increased significantly from at 24, 48 and 72 h post infection compared with 6 h post infection (P<0.05, 0.05 and 0.01, respectively) (Fig. 3B).

Bottom Line: The proliferation rate of T24 cells was significantly reduced by NDRG2 expression (P<0.01).In addition, 82.1% of NDRG2‑expressing cells were in S‑phase, compared to 74.4% in the control virus‑infected cells (P<0.05).In conclusion, the results of the present study indicated that NDRG2 expression in mitochondria may arrest bladder cancer cells in S‑phase as well as decrease cell proliferation through inducing oncosis.

View Article: PubMed Central - PubMed

Affiliation: Cancer Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong 524001, P.R. China.

ABSTRACT
Previous studies have reported the antitumor activity of N‑Myc downstream‑regulated gene 2 (NDRG2), a novel p53‑inducible gene, in several types of cancer. The present study aimed to investigate the effects of NDRG2 expression on the proliferation of a human bladder cancer cell line. NDRG2 and control green fluorescent protein (GFP) recombinant adenovirus plasmids were constructed and transfected into a bladder cancer cell line with mutant p53 (T24 cells). NDRG2 expression was analyzed using western blot analysis and immunofluorescence assay (IFA); in addition, the subcellular localization of NDRG2 was detected using a confocal microscope. The proliferation rate of cells was measured using colony formation and MTT assays. Furthermore, the cell cycle of transfected T24 cells was detected by flow cytometry. The results indicated that T24 cells expressed low levels of NDRG2 prior to infection with GFP‑NDRG2 recombinant adenovirus; by contrast, following infection, NDRG2 was primarily overexpressed in mitochondria. The proliferation rate of T24 cells was significantly reduced by NDRG2 expression (P<0.01). In addition, 82.1% of NDRG2‑expressing cells were in S‑phase, compared to 74.4% in the control virus‑infected cells (P<0.05). Furthermore, upregulation of NDRG2 induced an increase in oncosis, rather than apoptosis, in T24 cell. In conclusion, the results of the present study indicated that NDRG2 expression in mitochondria may arrest bladder cancer cells in S‑phase as well as decrease cell proliferation through inducing oncosis. It was therefore proposed that NDRG2 was not only a biomarker, but also a tumor suppressor for bladder cancer.

Show MeSH
Related in: MedlinePlus