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STAT1 modification improves therapeutic effects of interferons on lung cancer cells.

Chen J, Zhao J, Chen L, Dong N, Ying Z, Cai Z, Ji D, Zhang Y, Dong L, Li Y, Jiang L, Holtzman MJ, Chen C - J Transl Med (2015)

Bottom Line: We found that expression of STAT1 or STAT1-CC enhanced the effect of IFN-γ and, IFN-β on inhibition of human lung cancer cell proliferation, migration and invasiveness.STAT1-CC showed increased effects compared to STAT1 on IFN-γ induced pSTAT1 and down-regulation of S100A4, PCNA, and c-fos levels.The results show that STAT1-CC exhibited more strength in improving the antitumor response of IFNs in lung cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang, China. jiestory414@163.com.

ABSTRACT

Background: Interferons (IFNs) have potent anti-proliferative, pro-apoptotic, and immunomodulatory activities against cancer. However, the clinical utility of IFNs is limited by toxicity and pharmacokinetics making it difficult to achieve sustained therapeutic levels especially in solid tumors.

Methods: Signal Transducer and Activator of Transcription 1 (STAT1) or a modified STAT1 (designated STAT1-CC) that is hyper-responsive to IFN were overexpressed in lung cancer SPC-A-1 and H1299 cells using lentiviral vectors. Transduction efficiency was monitored using enhanced green fluorescent protein (EGFP) expression. After transduction, cells were treated with interferon-gamma (IFN-γ) or interferon-beta (IFN-β) and monitored for cell proliferation, migration, and invasiveness using Cell Counting Kit-8 and transwell chamber assays and for apoptosis using Annexin V detection by flow cytometry. In addition, levels of STAT1, STAT1 Tyr-701 phosphorylation (pSTAT1), fibronectin, and β-catenin were determined using western blotting. In the case of IFN-γ stimulation, levels of S100A4, proliferating cell nuclear antigen (PCNA), and c-fos expression were also determined.

Results: We found that expression of STAT1 or STAT1-CC enhanced the effect of IFN-γ and, IFN-β on inhibition of human lung cancer cell proliferation, migration and invasiveness. Moreover, STAT1 and STAT1-CC expression caused increases in pSTAT1 and decreases in fibronectin and β-catenin levels. STAT1-CC showed increased effects compared to STAT1 on IFN-γ induced pSTAT1 and down-regulation of S100A4, PCNA, and c-fos levels.

Conclusion: The results show that STAT1-CC exhibited more strength in improving the antitumor response of IFNs in lung cancer cells. Results from this study suggest that combined treatment of IFNs and STAT1-CC might be a feasible approach for the clinical management of lung cancer in the future.

No MeSH data available.


Related in: MedlinePlus

Overexpressing STAT1-CC enhances IFN-γ or IFN-β induced apoptosis of lung cancer cells. Apoptosis was detected by Annexin V apoptosis detection Kit and analysed by flow cytometry. a Flow cytometry results were demonstrated from control, EGFP-, STAT1-, and STAT1-CC-expressing SPC-A-1 cells treated with IFN-γ 10,000 U/ml or IFN-β 10,000 U/ml for 96 h. b Quantitative analysis of the population of SPC-A-1 apoptotic cells. c Control, EGFP-, STAT1-, and STAT1-CC-expressing H1299 cells were treated with IFN-γ 1000 U/ml or IFN-β 1000 U/ml for 48 h. Quantitative analysis of the population of apoptotic cells in Three independent experiments were conducted and data are shown as mean ± SEM. **P < 0.01
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Fig5: Overexpressing STAT1-CC enhances IFN-γ or IFN-β induced apoptosis of lung cancer cells. Apoptosis was detected by Annexin V apoptosis detection Kit and analysed by flow cytometry. a Flow cytometry results were demonstrated from control, EGFP-, STAT1-, and STAT1-CC-expressing SPC-A-1 cells treated with IFN-γ 10,000 U/ml or IFN-β 10,000 U/ml for 96 h. b Quantitative analysis of the population of SPC-A-1 apoptotic cells. c Control, EGFP-, STAT1-, and STAT1-CC-expressing H1299 cells were treated with IFN-γ 1000 U/ml or IFN-β 1000 U/ml for 48 h. Quantitative analysis of the population of apoptotic cells in Three independent experiments were conducted and data are shown as mean ± SEM. **P < 0.01

Mentions: IFNs have potent antiproliferative and apoptosis inducing functions in several types of cancer cells. To examine whether overexpression of STAT1 or STAT1-CC will affect apoptotic events in the transduced lung cancer cells, apoptosis was measured using Annexin V apoptosis assay. STAT1 or STAT1-CC induced apoptosis in transduced SPC-A-1 cells under normal culture conditions. More importantly, STAT1- or STAT1-CC-expressing lung cancer SPC-A-1 cells showed increased levels of apoptosis compared to parent and EGFP expressing cells when treated with either IFN-γ (10,000 U/ml) or IFN-β (10,000 U/ml) for 96 h (Fig. 5). Notably, when treated with IFN-γ or IFN-β, SPC-A-1 cells transduced with STAT1-CC exhibited a significantly increased rate of apoptosis compared to cells transduced with wild type STAT1.Fig. 5


STAT1 modification improves therapeutic effects of interferons on lung cancer cells.

Chen J, Zhao J, Chen L, Dong N, Ying Z, Cai Z, Ji D, Zhang Y, Dong L, Li Y, Jiang L, Holtzman MJ, Chen C - J Transl Med (2015)

Overexpressing STAT1-CC enhances IFN-γ or IFN-β induced apoptosis of lung cancer cells. Apoptosis was detected by Annexin V apoptosis detection Kit and analysed by flow cytometry. a Flow cytometry results were demonstrated from control, EGFP-, STAT1-, and STAT1-CC-expressing SPC-A-1 cells treated with IFN-γ 10,000 U/ml or IFN-β 10,000 U/ml for 96 h. b Quantitative analysis of the population of SPC-A-1 apoptotic cells. c Control, EGFP-, STAT1-, and STAT1-CC-expressing H1299 cells were treated with IFN-γ 1000 U/ml or IFN-β 1000 U/ml for 48 h. Quantitative analysis of the population of apoptotic cells in Three independent experiments were conducted and data are shown as mean ± SEM. **P < 0.01
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4562290&req=5

Fig5: Overexpressing STAT1-CC enhances IFN-γ or IFN-β induced apoptosis of lung cancer cells. Apoptosis was detected by Annexin V apoptosis detection Kit and analysed by flow cytometry. a Flow cytometry results were demonstrated from control, EGFP-, STAT1-, and STAT1-CC-expressing SPC-A-1 cells treated with IFN-γ 10,000 U/ml or IFN-β 10,000 U/ml for 96 h. b Quantitative analysis of the population of SPC-A-1 apoptotic cells. c Control, EGFP-, STAT1-, and STAT1-CC-expressing H1299 cells were treated with IFN-γ 1000 U/ml or IFN-β 1000 U/ml for 48 h. Quantitative analysis of the population of apoptotic cells in Three independent experiments were conducted and data are shown as mean ± SEM. **P < 0.01
Mentions: IFNs have potent antiproliferative and apoptosis inducing functions in several types of cancer cells. To examine whether overexpression of STAT1 or STAT1-CC will affect apoptotic events in the transduced lung cancer cells, apoptosis was measured using Annexin V apoptosis assay. STAT1 or STAT1-CC induced apoptosis in transduced SPC-A-1 cells under normal culture conditions. More importantly, STAT1- or STAT1-CC-expressing lung cancer SPC-A-1 cells showed increased levels of apoptosis compared to parent and EGFP expressing cells when treated with either IFN-γ (10,000 U/ml) or IFN-β (10,000 U/ml) for 96 h (Fig. 5). Notably, when treated with IFN-γ or IFN-β, SPC-A-1 cells transduced with STAT1-CC exhibited a significantly increased rate of apoptosis compared to cells transduced with wild type STAT1.Fig. 5

Bottom Line: We found that expression of STAT1 or STAT1-CC enhanced the effect of IFN-γ and, IFN-β on inhibition of human lung cancer cell proliferation, migration and invasiveness.STAT1-CC showed increased effects compared to STAT1 on IFN-γ induced pSTAT1 and down-regulation of S100A4, PCNA, and c-fos levels.The results show that STAT1-CC exhibited more strength in improving the antitumor response of IFNs in lung cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang, China. jiestory414@163.com.

ABSTRACT

Background: Interferons (IFNs) have potent anti-proliferative, pro-apoptotic, and immunomodulatory activities against cancer. However, the clinical utility of IFNs is limited by toxicity and pharmacokinetics making it difficult to achieve sustained therapeutic levels especially in solid tumors.

Methods: Signal Transducer and Activator of Transcription 1 (STAT1) or a modified STAT1 (designated STAT1-CC) that is hyper-responsive to IFN were overexpressed in lung cancer SPC-A-1 and H1299 cells using lentiviral vectors. Transduction efficiency was monitored using enhanced green fluorescent protein (EGFP) expression. After transduction, cells were treated with interferon-gamma (IFN-γ) or interferon-beta (IFN-β) and monitored for cell proliferation, migration, and invasiveness using Cell Counting Kit-8 and transwell chamber assays and for apoptosis using Annexin V detection by flow cytometry. In addition, levels of STAT1, STAT1 Tyr-701 phosphorylation (pSTAT1), fibronectin, and β-catenin were determined using western blotting. In the case of IFN-γ stimulation, levels of S100A4, proliferating cell nuclear antigen (PCNA), and c-fos expression were also determined.

Results: We found that expression of STAT1 or STAT1-CC enhanced the effect of IFN-γ and, IFN-β on inhibition of human lung cancer cell proliferation, migration and invasiveness. Moreover, STAT1 and STAT1-CC expression caused increases in pSTAT1 and decreases in fibronectin and β-catenin levels. STAT1-CC showed increased effects compared to STAT1 on IFN-γ induced pSTAT1 and down-regulation of S100A4, PCNA, and c-fos levels.

Conclusion: The results show that STAT1-CC exhibited more strength in improving the antitumor response of IFNs in lung cancer cells. Results from this study suggest that combined treatment of IFNs and STAT1-CC might be a feasible approach for the clinical management of lung cancer in the future.

No MeSH data available.


Related in: MedlinePlus