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Anti-tumor activity of CIK cells on cervical cancer HeLa cells in vitro and in vivo and their distribution characteristics in tumor-bearing mice
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Abstract:
Objective ：To study the anti-tumor activity of cytokine-induced killer cells (CIK cells) against HeLa-luc cells (cervical cancer HeLa cells labeling luciferase) in vivo and in vitro, and to analyze the distribution characteristics of CIK cells in different organs in a mouse xenograft model of cervical cancer nude. Methods: CIK cells were induced from peripheral blood mononuclear cells of health volunteers and cultured in vitro. The phenotype of CIK cells were determined by flow cytometry. The expression of IFN-γ mRNA in CIK cells was detected by RT-PCR assay. The killing activity of CIK cells on HeLa-luc cells was determined by MTT assay and Wright-Giemsa’s staining. HeLa-luc cell-bearing BALB/c nude mouse model was established. Tumor size changes and treatment effect were detected using in vivo Xenogen IVIS Imaging System. The distribution characteristics of CIK cells in different organs were detected by confocal microscopy. Results: CIK cells grew up to the peak after being cultured for 14-21 d. The percentage of CD3+CD56+T cells was increased more 50 times than that of the beginning. The expression level of IFN-γ mRNA in CIK cells was also increased to the peak. When the ratios of effect to target were 20 ∶1, and 40 ∶1, the cytotoxicity of CIK cells on HeLa-luc cells was (5116±2.64)% and (72.14%±4.21)%, respectively, and was significantly higher than that of the control group (\[1633±3.09\]%, \[21.26±2.71\]%, respectively, P<0.05). The inhibitory rate of CIK cells on the tumor was 47.18% and 64.38% at the fifth week and the eighth week, respectively. The level of IFN-γ mRNA in the CIK experiment group (61.92±6.49) pg/ml was significantly higher than that in the control group (34.30±1.78) pg/ml (P<005). Green fluorescence can be observed in the lung, liver, spleen, peripheral blood and tumor tissues under the confocal microscope 3 h after CFSE-labeled CIK cells injection via peritoneal cavity and tumor adjacent. 24 h after injection via peritoneal cavity, the highest concentration of CIK cells was 20.56% in the tumor tissues, and 3 h after injection via tumor adjacent, the highest concentration of CIK cells was 25.75% in the tumor tissues. Conclusion: CIK cells show a strong cytotoxicity on cervical cancer HeLa-luc cells in vivo and in vitro. The CIK cells are extensively distributed into different organs after injection via peritoneal cavity or tumor adjacent. The concentration of CIK cells in different organ is closely related to the injection route and time.
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Project Supported:
Project supported by the Science and Technology Projects of Hebei Province (No. 09276101D-29), and the Key Specific Discipline Foundation of Higher Institutions of Hebei Province (No. 200552)
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