Effect of cells derived from tumor-draining lymph nodes of lung cancer patients on the expressions of apoptosis-related proteins in autologous tumor cells
- Author:
Ming HE
1
Author Information
1. Department of Chest Surgery
- Publication Type:Journal Article
- Keywords:
Apoptosis regulatory proteins, apotosis;
Carcinoma, squamous cell;
Lymph, nodes;
Mice, nude
- From:
Tumor
2007;27(7):562-565
- CountryChina
- Language:Chinese
-
Abstract:
Objectives: To study the effect of cells derived from tumor-draining lymph nodes (TDLN) of lung cancer patients on the expressions of apoptosis-related proteins in autologous tumor cells. Methods: Two to four tumor-draining lymph nodes without metastasis were collected from lung cancer patients during surgery. The TDLN cells were suspended in RPMI-1640 medium and stimulated with IL-2 175 U/mL (I-TDLN group) or IL-2 175 U/mL + GM-CSF + IL-4 500 U/mL (D-TDLN group) in culture flask, and then incubated at 37°C with 5% CO2. The human lung cancer cells were subcutaneously implanted into nude mice to establish tumor xenograft model. Then the nude mice were treated with TDLNs. The apoptotic rate and the expressions of Fas/FasL, Bax, Bcl-2, and survivin protein in xenograft tumors were analyzed by flow cytometry analysis. Results: The apoptotic rate of lung cancer cells was (6.07 ± 3.31)%, (12.11 ± 1.19)%, (21.42 ± 1.82)%, (24.60 ± 0.96)%, and (32.76 ± 4.46)% in blank control group, IL-2 group, LAK cells group, I-TDLN group, and G-TDLN group, respectively. ANOVA analysis showed there was significant difference in apoptotic rates between each group (P < 0.001). The fluorescence intensity (FI) of Fas protein was 1.46 ± 0.70, 1.47 ± 0.72, 1.44 ± 0.43, 1.42 ± 0.30, and 1.40 ± 0.45 in blank control group, IL-2 group, LAK cells group, I-TDLN group and G-TDLN group, respectively (P = 0.246). The FI value of FasL protein was 1.53 ± 0.29, 1.56 ± 0.41, 1.58 ± 0.56, 1.56 ± 0.36 and 1.57 ± 0.81 in blank control group, IL-2 group, LAK cells group, I-TDLN group, and G-TDLN group, respectively. The difference was not significant (P = 0. 528). The FI value of Bax protein was significantly higher in LAK group, I-TDLN group and G-TDLN than that in blank control and IL-2 group in blank control group (1.66 ± 0.84, 1.72 ± 0.53 and 1.87 ± 0.55 vs 1.36 ± 0.39 and 1.37 ± 0.16, respectively, P < 0. 001). The FI value of Bcl-2 protein was significantly lower in IL-2 group, LAK cells group, I-TDLN group, and G-TDLN group than that of blank control (1.39 ± 0.40, 1.22 ± 0.14, 1.15 ± 0.19 and 1.06 ± 0.75 vs 1.70 ± 0.88, respectively, P < 0.001). The FI value of survivin protein was significantly lower in LAK group, I-TDLN group and G-TDLN than that in blank control and IL-2 group (1.54 ± 0.62, 1.45 ± 0.61 and 1.35 ± 0.36 vs 1.71 ± 0.71 and 1.69 ± 0.20, respectively, P < 0.001). Conclusion: The autologous tumor-specific killing effect of TDLN cells is related with regulation of apoptosis-related proteins.
