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TGF-β1 Secreted by Hepatocellular Carcinoma Induces the Expression of the Foxp3 Gene and Suppresses Antitumor Immunity in the Tumor Microenvironment

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Abstract

Aim

The purpose of this study was to explore the mechanisms of TGF-β1 mediated immunosuppression in tumor stroma.

Methods

The expression of TGF-β1 was investigated in Huh7, Hep 3B, SGC-7901, Eca-109 and Hepa1-6 cell lines using immunofluorescence. Knocked-down TGF-β1 of the Hepa1-6 cell line was established through lentivirus-based RNA interference. The interference efficiency of the TGF-β1 gene was tested by real-time PCR and ELISA; the expression of Foxp3, IFN-γ and CD83 in CD4+, CD8+ or dendritic cells was examined via flow cytometry; and the tumorigenic ability of the cancer cells was investigated in the animal experiments.

Results

The diverse digestive cancer cells were found to secrete TGF-β1, mRNA of which was knocked down by 78 % thanks to lentivirus-based interference in Hepa1-6 cells. Flow cytometry showed that CD4+CD25+Foxp3+ regulatory T cells significantly increased in hepatocellular carcinoma patients when compared with those in the healthy controls. The supernatant from Hepa1-6 cells and recombinant TGF-β1 significantly induced the expression of Foxp3 gene in vitro, while that from sh TGF-β1 Hepa1-6 cells restored it. Hepa1-6 cells inhibited IFN-γ and CD83 expression in CD8+ or dendritic cells by secreting TGF-β1. The animal experiments indicated that the knockdown TGF-β1 gene impaired the tumorigenic ability of Hepa1-6 cells.

Conclusion

TGF-β1, expressed in cancer cells, might be a potential therapeutic target for cancer treatment.

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Acknowledgments

The authors would like to thank the team members of Prof. **zhong Shen's laboratory for their helpful discussion and critical reading of the manuscript. This study was supported by Shanghai Science and Technology Commission (10410709400; 10411950100) and National Nature Science Foundation of China (No. 30872503; No. 81000968; No. 81101540; No. 81172273), the National Clinical Key Special Subject of China.

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Authors and Affiliations

  1. The Department of Gastroenterology of Zhongshan Hospital, Fudan University, 180 Fenglin Rd, Shanghai, 200032, People’s Republic of China

    Yi Wang, Wenqing Tang, Taotao Liu & **zhong Shen

  2. The Department of Gastroenterology, Yangzhou First Hospital Affiliated to Yangzhou University, Yangzhou University, Yangzhou, Jiangsu, People’s Republic of China

    Bin Deng

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  1. Yi Wang
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Correspondence to **zhong Shen.

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Wang, Y., Deng, B., Tang, W. et al. TGF-β1 Secreted by Hepatocellular Carcinoma Induces the Expression of the Foxp3 Gene and Suppresses Antitumor Immunity in the Tumor Microenvironment. Dig Dis Sci 58, 1644–1652 (2013). https://doi.org/10.1007/s10620-012-2550-4

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