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Archive > Volume 24 Issue 4 > 2017,24(4):404-410. DOI:10.3872/j.issn.1007-385X.2017.04.012 Prev Next
Basic Research
Anti-tumor activity of cytokine-induced killer cells combined with bevacizumab against hepatocellular carcinoma HepG2 cells in vitro and in vivo
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Abstract:
Objective:To investigate the antitumor activity and the mechanism of cytokine-induced killer (CIK) cells combined with bevacizumab against hepatocellular carcinoma HepG2 cells in vitro and in vivo. Methods: Peripheral blood mononuclear cells (PBMCs) from healthy blood donors were isolated and cultured in the presence of different cytokines to promote the maturation of CIK cells; and the immune phenotypes of CIK cells were analyzed by flow cytometry. The inhibitory effect on HepG2 cells of CIK cells combining with or without bevacizumab were analyzed by CCK-8 assay. Cell migration and invasion of HepG2 cells were detected by scratch assay and transwell assay. The phosphorylation levels of Erk/Akt signaling pathway related proteins were detected by Western blotting. HepG2 cell xenograft tumor model was established in nude mice. Tumor bearing mice were divided into normal saline control group, CIK group, bevacizumab group and CIK+bevacizumab group. On the 28th day after drug administration, the mice were sacrificed and tumors were isolated. Immunohistochemical staining was used to analyze the expressions of CD31 and Ki67. Results: Human PBMCs were stimulated for 14 days, and the CIK cell phenotype analysis showed that the amplification of CD3+CD56+ CIK cells reached (36.33±2.58)%. The inhibition rate of CIK cells combined with bevacizumab was significantly enhanced by comparing with CIK/bevacizumab alone groups (all P<0.05). CIK cells combined with bevacizumab showed a more intensive inhibition on cell migration(\[75.6±9.53\] vs \[304.8±45.73\], \[359.8±38.10\], P<0.01) and invasion (\[29.35±8.14\]% vs \[55.07±6.27\]%, \[60.50±9.73\]%, P<0.05\] by comparing with those two single treatment groups; CIK cell treatment, bevacizumab treatment and the combined treatment could all decrease the phosphorylation level of Erk and Akt; the combined treatment significantly inhibited the tumor growth, mean vessel density (MVD) and Ki67 expression, which had statistical significance when compared with the other groups (P<0.05). Conclusion: The current study suggested that CIK cells combined with bevacizumab inhibited HepG2 cells proliferation, invasion and migration both in vitro and in vivo, which significantly precede the treatment efficacy of single treatment groups.
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