Immune Netw.  2012 Apr;12(2):66-69. 10.4110/in.2012.12.2.66.

Chemotherapeutic Candidate Inducing Immunological Death of Human Tumor Cell Lines

• Oh SJ
• Ryu CK
• Choi I
• Baek SY
• Lee H

• KMID: 2150736
• DOI: http://doi.org/10.4110/in.2012.12.2.66

Abstract

The immunological death induction by EY-6 on the human tumor cell lines was screened. Human colon carcinoma (HCT15, HCT116), gastric carcinoma (MKN74, SNU668), and myeloma (KMS20, KMS26, KMS34) cells were died by EY-6 treatment with dose-dependent manner. CRT expression, a typical marker for the immunological death, was increased on the EY-6-treated colorectal and gastric cancer cells. Interestingly, the effects on the myeloma cell lines were complicated showing cell line dependent differential modulation. Cytokine secretion from the EY-6 treated tumor cells were dose and cell-dependent. IFN-gamma and IL-12 secretion was increased in the treated cells (200% to over 1000% of non-treated control), except HCT116, SNU668 and KMS26 cells which their secretion was declined by EY-6. Data suggest the potential of EY-6 as a new type of immuno-chemotherapeutics inducing tumor-specific cell death. Further studies are planned to confirm the efficacy of EY-6 including in vivo study.

Keyword

Indoledione derivative; Calreticulin; Immunological death; Human tumor cells

MeSH Terms

Calreticulin
Cell Death
Cell Line
Cell Line, Tumor
Colon
Humans
Interleukin-12
Stomach Neoplasms
Calreticulin
Interleukin-12

Figure

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  Figure 1 Cytotoxicity induced by EY-6 was determined by MTT assay. Human tumor cell lines (colon cancer cells HCT15, HCT116; gastric cancer cells MKN74, SNU668; and myeloma cells KMS20, KMS26, KMS34) were incubated with different doses (0, 25 and 50µM) of EY-6 for 48 hr at 37℃. After the incubation, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT solution, 5 mg/ml) was added and incubated for 4 h at 37℃. At the end of the incubation, supernatant was removed and the color change induced by dimethyl sulfoxide (DMSO) was determined at 540 nm with ELISA reader (Molecular Devices, Sunnyvale, CA, USA). The analyzed values were reported as the mean±standard error. Statistical significance was determined by two-tailed Student's t-test. Asterisks represented the statistical significance (p<0.05) compared to 0µM-control group.

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  Figure 2 EY-6 induced IL-12 secretion from the tumor cells. Culture supernatants from tumor cells incubated with different doses of EY-6 for 24 hr were obtained to measure the secreted IL-12 by ELISA using commercially available assay kit (eBioscience, SanDiego, CA, USA). The amount of IL-12 (pg/ml) was reported as the mean±standard error. Statistical significance was determined by two-tailed Student's t-test. Asterisks represented the statistical significance (p<0.05) compared to control group.

• 

  Figure 3 EY-6 induced IFN-γ secretion from the tumor cells. Culture supernatants from tumor cells incubated with different doses of EY-6 for 24 hr were obtained to measure the secreted IFN-γ by ELISA using commercially available assay kit (eBioscience, SanDiego, CA, USA). The amount of IFN-γ (pg/ml) was reported as the mean±standard error. Statistical significance was determined by two-tailed Student's t-test. Asterisks represented the statistical significance (p<0.05) compared to control group.

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