Journal: Oncology Letters

Article Title: MicroRNA-374b inhibits liver cancer progression via down regulating programmed cell death-1 expression on cytokine-induced killer cells

doi: 10.3892/ol.2018.7951

Figure Lengend Snippet: Effect of PD-1 on IFN-γ production, cell viability, cytotoxicity and antitumor effect in CIK cells in vitro and in vivo . A synthetic siRNA targeting PD-1 was employed to silence the expression level of PD-1 in CIK cells. (A) An ELISA was used to measure the expression of IFN-γ in CIK cells. The bands were quantified (**P<0.01 vs. CIK+Inhibitor+NC group). (B) ALDH release assay was used to determine the cytotoxicity of the CIK on HepG2 cells (**P<0.01 vs. CIK+Inhibitor+NC+HepG2 group). (C) The Cell Counting Kit-8 assay was performed to visualize the proliferation of HepG2 cells with CIK (**P<0.01 vs. CIK+Inhibitor+NC+HepG2 group). (D) Winn assay and (E) adoptive transfer assay were used to examine the antitumor effect of siPD-1 transfected CIK cells in nude mice. The volume of the tumors in different groups is depicted (*P<0.05,**P<0.01 vs. CIK+Inhibitor+NC+HepG2 group). IFN, interferon; siPD-1, small interfering programmed cell death-1; NC, negative control; CIK, cytokine-induced killer; Inhibitor, miR-374b inhibitor; LDH, lactate dehydrogenase; OD, optical density.

Article Snippet: Finally, 50 μl stop solution (50% dimethyl form amide and 20% sodium dodecyl sulfate; pH 4.7) was added and the concentration of LDH in each well was measured using an automatic ELISA reader (ELISA reader; ASYS Hitech, GmbH, Austria).

Techniques: In Vitro, In Vivo, Expressing, Enzyme-linked Immunosorbent Assay, Release Assay, Cell Counting, Adoptive Transfer Assay, Transfection, Negative Control