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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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Validation of transfection efficiency in T24 cells using fluorescence microscopy. (A) Representative images of GFP expression in T24, T24-GFP and T24-NDRG2 cells. (B) Quantification of GFP gene transfection efficiency. Values are presented as the mean ± standard deviation. *P<0.01 vs. T24 cells. GFP, green fluorescent protein; T24-GFP, control GFP adenovirus-infected T24 cells; T24-NDRG2, GFP-N-Myc downstream-regulated gene 2 adenovirus-infected T24 cells.
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f1-mmr-12-04-5730: Validation of transfection efficiency in T24 cells using fluorescence microscopy. (A) Representative images of GFP expression in T24, T24-GFP and T24-NDRG2 cells. (B) Quantification of GFP gene transfection efficiency. Values are presented as the mean ± standard deviation. *P<0.01 vs. T24 cells. GFP, green fluorescent protein; T24-GFP, control GFP adenovirus-infected T24 cells; T24-NDRG2, GFP-N-Myc downstream-regulated gene 2 adenovirus-infected T24 cells.

Mentions: In order to validate the success of infection, cell morphology was observed using a light microscope and fluorescence microscope. The results showed that ~90% cells expressed GFP in the pAV.EX1d-eGFP-infected group; by contrast, only ~60% cells expressed GFP in the pAV. EX1d-NDRG2/IRES/eGFP-infected group (Fig. 1A and B).


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)

Validation of transfection efficiency in T24 cells using fluorescence microscopy. (A) Representative images of GFP expression in T24, T24-GFP and T24-NDRG2 cells. (B) Quantification of GFP gene transfection efficiency. Values are presented as the mean ± standard deviation. *P<0.01 vs. T24 cells. GFP, green fluorescent protein; T24-GFP, control GFP adenovirus-infected T24 cells; T24-NDRG2, GFP-N-Myc downstream-regulated gene 2 adenovirus-infected T24 cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1-mmr-12-04-5730: Validation of transfection efficiency in T24 cells using fluorescence microscopy. (A) Representative images of GFP expression in T24, T24-GFP and T24-NDRG2 cells. (B) Quantification of GFP gene transfection efficiency. Values are presented as the mean ± standard deviation. *P<0.01 vs. T24 cells. GFP, green fluorescent protein; T24-GFP, control GFP adenovirus-infected T24 cells; T24-NDRG2, GFP-N-Myc downstream-regulated gene 2 adenovirus-infected T24 cells.
Mentions: In order to validate the success of infection, cell morphology was observed using a light microscope and fluorescence microscope. The results showed that ~90% cells expressed GFP in the pAV.EX1d-eGFP-infected group; by contrast, only ~60% cells expressed GFP in the pAV. EX1d-NDRG2/IRES/eGFP-infected group (Fig. 1A and B).

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