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. 2005 Jun 7;11(21):3204-11.
doi: 10.3748/wjg.v11.i21.3204.

Anti-gastric cancer active immunity induced by FasL/B7-1 gene-modified tumor cells

Shi-Ying Zheng  1 , De-Chun Li, Zhi-De Zhang, Jun Zhao, Jin-Feng Ge
Affiliations

Anti-gastric cancer active immunity induced by FasL/B7-1 gene-modified tumor cells

Shi-Ying Zheng et al. World J Gastroenterol. .

Abstract

Aim: To study the activation of cytotoxic T lymphocytes (CTLs) against gastric cancer cells induced by FasL/B7-1 (FB-11) gene-modified tumor cells, and to explore whether co-expression of FasL and B7-1 in SGC-7901 tumor cells could initiate synergistic antitumor effect.

Methods: FasL and B7-1 genes were transfected into human SGC-7901 gastric cancer cells with adenovirus vectors. The positive clones were selected by G418. FasL and B7-1 genes were detected by flow cytometry and RT-PCR. Abdominal infiltrating lymphocytes and sensitized spleen cells were obtained from mice that were immunized with SGC-7901/FB-11 or wild type SGC-7901 cells intraperitoneally, and cytotoxicity of these CTLs against tumor cells was determined by MTT assay.

Results: Flow cytometry and RT-PCR showed that FasL and B7-1 genes were highly expressed. FasL and B7-1 transfected cancer cells had a high apoptosis index. DNA laddering suggested that FasL and B7-1 genes induced gastric cancer cell apoptosis. FasL(+)/B7-1(+)SGC-7901 cells (SGC-7901/FB-11) were inoculated subcutaneously in the dorsal skin of C57BL/6 mice and then decreased their tumorigenicity greatly (z = 2.15-46.10, P<0.01). SGC-7901/FB-11 cell-sensitized mice obtained protective immune activity against the rechallenge of wild type SGC-7901 cells (z = 2.06-44.30, P<0.05). The cytotoxicity of CTLs induced by SGC-7901/FB-11 cells against SGC-7901 was significantly higher than that of CTLs activated by wild-type SGC-7901 cells (84.1+/-2.4% vs 30.5+/-2.3%, P<0.05).

Conclusion: FasL and B7-1 genes can effectively promote the activity of CTLs against gastric cancer cells. FasL/B7-1 molecules play an important role in CTL cytotoxicity.

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Figures

Figure 1
Figure 1
Transfection rates of rAd in gastric cancer cells. A: MOI = 20; B: MOI = 50; C: MOI = 100.
Figure 2
Figure 2
Flow cytometric analysis of B7-1 gene expression. A: SGC-7901; B: SGC-7901/FB-11.
Figure 3
Figure 3
RT-PCR analysis of FasL expression in SGC-7901/FB-11 and SGC-7901. A: SGC-7901/FB-11; B: SGC-7901; C: DNA Marks.
Figure 4
Figure 4
Proliferation inhibition of gastric cancer cell lines. ■: SGC-7901; ○: SGC-7901L/FasL; ●: SGC-7901/FB-11; □: PBS.
Figure 5
Figure 5
Apoptosis of SGC-7901 cells stained with Hoechst 33342 induced by FB-11 (×400). A: SGC-7901/GFP cells; B: SGC-7901/FB-11 cells.
Figure 6
Figure 6
DNA fragmentation of SGC-7901 cells induced by FB-11. A: DNA Marks; B: SGC-7901/FB-11; C: MGC-803/FB-11; D: BGC-823/FB-11; E: SGC-7901.
Figure 7
Figure 7
Tumorigenicity of FB-11-transfected gastric cancer cells and tumor size in preventive and control groups. ●: FB-11 preventive group; ■: control group.
Figure 8
Figure 8
FB-11-mediated protective immunity. ■: SGC-7901/FB-11-sensitized mice group; ●: SGC-7901 mice group (not sensitized with SGC-7901/FB-11).
Figure 9
Figure 9
Cytotoxicity of abdominal lymphocyte infiltration against tumor cells induced by FB-11. A: cytotoxicity of SGC-7901/FB-11-induced abdominal lymphocyte infiltration against SGC-7901/FB-11 cells (n = 4); B: cytotoxicity of SGC-7901/FB-11-induced abdominal lymphocyte infiltration against SGC-7901 cells (n = 4); C: cytotoxicity of SGC-7901-induced abdominal lymphocyte infiltration against SGC-7901 cells (n = 4).
Figure 10
Figure 10
Cytotoxicity of spleen cells against tumor cells induced by FB-11. A: cytotoxicity of SGC-7901/FB-11-induced spleen cells against SGC-7901/FB-11 cells (n = 4); B: Cytotoxicity of SGC-7901/FB-11-induced spleen cells against SGC-7901 cells (n = 4); C: Cytotoxicity of SGC-7901-induced spleen cells against SGC-7901 cells (n = 4).
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