T-cell apoptosis induced by intratumoral activated hepatic stellate cells is associated with lung metastasis in hepatocellular carcinoma

Xia,Yun-Hong; Wang,Zhi-Ming; Chen,Rong-Xin; Ye,Sheng-Long; Sun,Rui-Xia; Xue,Qiong; Huang,Yong

doi:10.3892/or.2013.2571

T-cell apoptosis induced by intratumoral activated hepatic stellate cells is associated with lung metastasis in hepatocellular carcinoma

Authors:
- Yun-Hong Xia
- Zhi-Ming Wang
- Rong-Xin Chen
- Sheng-Long Ye
- Rui-Xia Sun
- Qiong Xue
- Yong Huang
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Affiliations: Department of Oncology, Hefei Second People's Hospital, Anhui Medical University, Hefei 230011, P.R. China, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai 200032, P.R. China
Published online on: June 27, 2013 https://doi.org/10.3892/or.2013.2571
Pages: 1175-1184

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Abstract

Profound T cell inhibitory activity of hepatic stellate cells (HSCs) in vitro has recently been described in hepatocellular carcinoma (HCC). In the present study, we investigated the immune inhibitory activity of HSCs in vivo in an orthotopic rat HCC model with lung metastasis. Rats (n=24) were randomly sacrificed on days 7, 14, 21 and 28 (n=4 each). Lung tissues were stained with hematoxylin and eosin. Liver sections were stained for immunofluorescence analysis. T-cell apoptosis was detected using double staining for terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Staining revealed marked and continuous accumulation of α-smooth muscle actin with tumor progression after orthotopic tumor implantation in rat liver. T lymphocyte numbers gradually increased following tumor progression, and subset analysis revealed an increase in the distribution of liver CD8+ and CD4+ T cells. Double staining for CD3 and TUNEL demonstrated T-cell apoptosis. Apoptotic T cells were more frequent in the HCC livers compared to the normal livers, and were spatially associated with intratumoral activated HSCs (tHSCs), suggesting a direct interaction. T-cell apoptosis was more frequently induced in the co-cultures of activated splenic T cells(aT)/tHSCs compared to aT/quiescent (q) HSCs or qT/tHSCs. tHSCs were positively correlated with T-cell apoptosis, and the percentage of T-cells undergoing apoptosis was positively correlated with the number of lung metastasis nodules. T-cell apoptosis may be promoted via an interaction with tHSCs, suggesting that tHSCs regulate T cells and contribute to lung metastasis in HCC.

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