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CpG oligonucleotides suppress HepG2 cells-induced Jurkat cell apoptosis via the Fas-FasL-mediated pathway.

Zheng J, Fu R, Li J, Wang X - J. Exp. Clin. Cancer Res. (2011)

Bottom Line: To explore the potential role of CpG motif-containing oligonucleotides (CpG-ODN) in modulating the expression of FasL in HepG2 and Fas in Jurkat cells in vitro, and to examine the effect of CpG-ODN treatment on the HepG2 cells-mediated Jurkat cell apoptosis in vitro.Pre-treatment of HepG2 or Jurkat cells with CpG-ODN significantly reduced the frequency of HepG2-mediated apoptotic Jurkat cells and inhibited the activation of caspase-3 in Jurkat cells in vitro.Apparently, CpG-ODN treatment may be a potential therapeutic reagent for HCC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Clinical Laboratory, the Second Affiliated hospital of Nanchang University, Nanchang 330006, China.

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Apoptosis of Jurkat cells induced by HepG2 cells. HepG2 and Jurkat cells were cultured in medium alone or treated with 1 μM CpG-ODN or 10 μg/ml xx μg/ml anti-FasL NOK-2 antibody for 24 h. The cells were harvested and co-cultured as the unmanipulated HepG2 and Jurkat cells (A, positive controls), the NOK-2-treated HepG2 and unmanipulated Jurkat cells (B), the unmanipulated HepG2 and NOK_2-treated Jurkat cells (C), the CpG-ODN-treated HepG2 and unmanipulated Jurkat cells (D) or the unmanipulated HepG2 and CpG-ODN-treated Jurkat cells (E), respectively for 24 h. The unadhered Jurkat cells were harvested and stained with FITC-Annexin V and PI, followed by flow cytometry analysis. (F) Quantitative analysis. The frequency of apoptotic Jurkat cells was analyzed by using CellQuest software. Data are expressed as representative FCM or mean% ± S.E.M of each group of the cells from four independent experiments. *p < 0.05 vs. the positive controls.
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Figure 3: Apoptosis of Jurkat cells induced by HepG2 cells. HepG2 and Jurkat cells were cultured in medium alone or treated with 1 μM CpG-ODN or 10 μg/ml xx μg/ml anti-FasL NOK-2 antibody for 24 h. The cells were harvested and co-cultured as the unmanipulated HepG2 and Jurkat cells (A, positive controls), the NOK-2-treated HepG2 and unmanipulated Jurkat cells (B), the unmanipulated HepG2 and NOK_2-treated Jurkat cells (C), the CpG-ODN-treated HepG2 and unmanipulated Jurkat cells (D) or the unmanipulated HepG2 and CpG-ODN-treated Jurkat cells (E), respectively for 24 h. The unadhered Jurkat cells were harvested and stained with FITC-Annexin V and PI, followed by flow cytometry analysis. (F) Quantitative analysis. The frequency of apoptotic Jurkat cells was analyzed by using CellQuest software. Data are expressed as representative FCM or mean% ± S.E.M of each group of the cells from four independent experiments. *p < 0.05 vs. the positive controls.

Mentions: Engagement of Fas on the cell membrane by FasL can trigger cell apoptosis. Given that CpG-ODN treatment down-regulated the expression of FasL in HepG2 cells and Fas in Jurkat cells, it is possible that CpG-ODN may modulate the HepG2 cell-mediated Jurkat cell apoptosis. Accordingly, we first treated HepG2 and Jurkat cells with 1 μM CpG-PDN or anti-FasL NOK-2 antibody for 24 h for the preparation of effector and target cells, respectively. Next, we co-cultured the unmanipulated HepG2 and Jurkat cells (positive controls), the NOK-2-treated HepG2 and untreated Jurkat cells, the untreated HepG2 and the NOK-2-treated Jurkat cells, the CpG-ODN-treated HepG2 and untreated Jurkat cells, and the untreated HepG2 and the CpG-ODN-treated Jurkat cells for 24, respectively. Subsequently, the suspended Jurkat cells were collected and the frequency of apoptotic Jurkat cells was determined by flow cytometry analysis (Figure 3). First, co-culture of HepG2 cells with Jurkat cells triggered Jurkat cell apoptosis (Figure 3A and 3F). Pre-treatment of either HepG2 or Jurkat cells with anti-FasL antibody significantly reduced the frequency of apoptotic Jurkat cells (Figure 3B and 3C), indicating that the FasL/Fas pathway might be involved in the apoptosis of Jurkat cells in this experimental system.


CpG oligonucleotides suppress HepG2 cells-induced Jurkat cell apoptosis via the Fas-FasL-mediated pathway.

Zheng J, Fu R, Li J, Wang X - J. Exp. Clin. Cancer Res. (2011)

Apoptosis of Jurkat cells induced by HepG2 cells. HepG2 and Jurkat cells were cultured in medium alone or treated with 1 μM CpG-ODN or 10 μg/ml xx μg/ml anti-FasL NOK-2 antibody for 24 h. The cells were harvested and co-cultured as the unmanipulated HepG2 and Jurkat cells (A, positive controls), the NOK-2-treated HepG2 and unmanipulated Jurkat cells (B), the unmanipulated HepG2 and NOK_2-treated Jurkat cells (C), the CpG-ODN-treated HepG2 and unmanipulated Jurkat cells (D) or the unmanipulated HepG2 and CpG-ODN-treated Jurkat cells (E), respectively for 24 h. The unadhered Jurkat cells were harvested and stained with FITC-Annexin V and PI, followed by flow cytometry analysis. (F) Quantitative analysis. The frequency of apoptotic Jurkat cells was analyzed by using CellQuest software. Data are expressed as representative FCM or mean% ± S.E.M of each group of the cells from four independent experiments. *p < 0.05 vs. the positive controls.
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Figure 3: Apoptosis of Jurkat cells induced by HepG2 cells. HepG2 and Jurkat cells were cultured in medium alone or treated with 1 μM CpG-ODN or 10 μg/ml xx μg/ml anti-FasL NOK-2 antibody for 24 h. The cells were harvested and co-cultured as the unmanipulated HepG2 and Jurkat cells (A, positive controls), the NOK-2-treated HepG2 and unmanipulated Jurkat cells (B), the unmanipulated HepG2 and NOK_2-treated Jurkat cells (C), the CpG-ODN-treated HepG2 and unmanipulated Jurkat cells (D) or the unmanipulated HepG2 and CpG-ODN-treated Jurkat cells (E), respectively for 24 h. The unadhered Jurkat cells were harvested and stained with FITC-Annexin V and PI, followed by flow cytometry analysis. (F) Quantitative analysis. The frequency of apoptotic Jurkat cells was analyzed by using CellQuest software. Data are expressed as representative FCM or mean% ± S.E.M of each group of the cells from four independent experiments. *p < 0.05 vs. the positive controls.
Mentions: Engagement of Fas on the cell membrane by FasL can trigger cell apoptosis. Given that CpG-ODN treatment down-regulated the expression of FasL in HepG2 cells and Fas in Jurkat cells, it is possible that CpG-ODN may modulate the HepG2 cell-mediated Jurkat cell apoptosis. Accordingly, we first treated HepG2 and Jurkat cells with 1 μM CpG-PDN or anti-FasL NOK-2 antibody for 24 h for the preparation of effector and target cells, respectively. Next, we co-cultured the unmanipulated HepG2 and Jurkat cells (positive controls), the NOK-2-treated HepG2 and untreated Jurkat cells, the untreated HepG2 and the NOK-2-treated Jurkat cells, the CpG-ODN-treated HepG2 and untreated Jurkat cells, and the untreated HepG2 and the CpG-ODN-treated Jurkat cells for 24, respectively. Subsequently, the suspended Jurkat cells were collected and the frequency of apoptotic Jurkat cells was determined by flow cytometry analysis (Figure 3). First, co-culture of HepG2 cells with Jurkat cells triggered Jurkat cell apoptosis (Figure 3A and 3F). Pre-treatment of either HepG2 or Jurkat cells with anti-FasL antibody significantly reduced the frequency of apoptotic Jurkat cells (Figure 3B and 3C), indicating that the FasL/Fas pathway might be involved in the apoptosis of Jurkat cells in this experimental system.

Bottom Line: To explore the potential role of CpG motif-containing oligonucleotides (CpG-ODN) in modulating the expression of FasL in HepG2 and Fas in Jurkat cells in vitro, and to examine the effect of CpG-ODN treatment on the HepG2 cells-mediated Jurkat cell apoptosis in vitro.Pre-treatment of HepG2 or Jurkat cells with CpG-ODN significantly reduced the frequency of HepG2-mediated apoptotic Jurkat cells and inhibited the activation of caspase-3 in Jurkat cells in vitro.Apparently, CpG-ODN treatment may be a potential therapeutic reagent for HCC.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Clinical Laboratory, the Second Affiliated hospital of Nanchang University, Nanchang 330006, China.

Show MeSH