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Effect of chaetocin on renal cell carcinoma cells and cytokine-induced killer cells.

Rombo R, Weiher H, Schmidt-Wolf IG - Ger Med Sci (2016)

Bottom Line: The addition of distinct interleukins and antibodies results in the generation of CIK cells that are able to specifically target and destroy renal carcinoma cells.Although the CIK cells show a high grade of selective anti-ccRCC activity, this effect could not be improved by the addition of chaetocin.Abstract available from the publisher.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Integrated Oncology (CIO), University of Bonn, Bonn, Germany; Hochschule Bonn-Rhein-Sieg, Rheinbach, Germany.

ABSTRACT
We examined the cytotoxic effects of chaetocin on clear cell renal cell carcinoma (ccRCC) cells and the possibility to combine the effects of chaetocin with the effects of cytokine-induced killer cells (CIK) assayed by MTT assay and FACS analysis. Chaetocin is a thiodioxopiperazine produced by fungi belonging to the chaetomiaceae family. In 2007, it was first reported that chaetocin shows potent and selective ex vivo anti-cancer activity by inducing reactive oxygen species. CIK cells are generated from CD3+/CD56- T lymphocytes with double negative CD4-/CD8- phenotype that are isolated from human blood. The addition of distinct interleukins and antibodies results in the generation of CIK cells that are able to specifically target and destroy renal carcinoma cells. The results of this research state that the anti-ccRCC activity of chaetocin is weak and does not show a high grade of selectivity on clear cell renal cell carcinoma cells. Although the CIK cells show a high grade of selective anti-ccRCC activity, this effect could not be improved by the addition of chaetocin. So chaetocin seems to be no suitable agent for specific targeting ccRCC cells or for the combination therapy with CIK cells in renal cancer.

No MeSH data available.


Related in: MedlinePlus

Results of the FACS analysis of ccRCC (A1, A2) and non-cancerous (B1, B2) cell line treated with CIK cells only (A1, B1) and CIK cells in combination with 200 nM chaetocin (A2, B2) for 24 h. Population of CIK cells and ccRCC cells were measured by FCS vs SSC. Hoechst 33258 stain and FSC was used to discriminate between living and dead ccRCC cells.
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Figure 5: Results of the FACS analysis of ccRCC (A1, A2) and non-cancerous (B1, B2) cell line treated with CIK cells only (A1, B1) and CIK cells in combination with 200 nM chaetocin (A2, B2) for 24 h. Population of CIK cells and ccRCC cells were measured by FCS vs SSC. Hoechst 33258 stain and FSC was used to discriminate between living and dead ccRCC cells.

Mentions: The ccRCC and CIK cells were combined in a ratio of 1:10 and incubated with and without chaetocin for 24 h. A concentration of 200 nM chaetocin was chosen since it killed 100% of the A-498 ccRCC cell line while only killing 28% of CCD-18Co and 25% of CIK. The MTT assay can be used to determine the cytotoxicity of a certain drug on a single cell line by simply stating how many cells survived the treatment. However, the MTT assay cannot be used to distinguish between two different cell lines when incubated together. In order to determine how many non-cancerous and ccRCC cells survived the incubation along with chaetocin and CIK cells a FACS was used. Forward scatter (FSC) and side scatter (SSC) were used to identify and discriminate CIK cells, non-cancerous and ccRCC cells. The Hoechst 33258 staining solution was used in order to stain dead cells independent of being cancerous or non-cancerous to see how many survive the treatment. It was only possible to test one non-cancerous and one ccRCC cell line due to a lack of resources. Figure 5A 1.1 (Fig. 5) shows the population of CIK cells and A-498 ccRCC cells after 24 h of incubation in the absence of chaetocin. Both populations could be discriminated by their SSC and FSC. Figure 5A 1.2 (Fig. 5) shows the ratio of dead and living A-498 cells discriminated by Hoechst vs FSC. After the treatment 50.6% of the ccRCC cells were still alive while 45.9 % died and 3.5% could not be matched to either group. Figure 5A 2.1 and A 2.2 (Fig. 5) show an identical experimental set up but this time the cells were incubated in the presence of 200 nM chaetocin. After 24 h of incubation along with chaetocin 56.4% of the ccRCC cells died while only 41.5% stayed alive and 2.1% could not be matched to either group.


Effect of chaetocin on renal cell carcinoma cells and cytokine-induced killer cells.

Rombo R, Weiher H, Schmidt-Wolf IG - Ger Med Sci (2016)

Results of the FACS analysis of ccRCC (A1, A2) and non-cancerous (B1, B2) cell line treated with CIK cells only (A1, B1) and CIK cells in combination with 200 nM chaetocin (A2, B2) for 24 h. Population of CIK cells and ccRCC cells were measured by FCS vs SSC. Hoechst 33258 stain and FSC was used to discriminate between living and dead ccRCC cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4836455&req=5

Figure 5: Results of the FACS analysis of ccRCC (A1, A2) and non-cancerous (B1, B2) cell line treated with CIK cells only (A1, B1) and CIK cells in combination with 200 nM chaetocin (A2, B2) for 24 h. Population of CIK cells and ccRCC cells were measured by FCS vs SSC. Hoechst 33258 stain and FSC was used to discriminate between living and dead ccRCC cells.
Mentions: The ccRCC and CIK cells were combined in a ratio of 1:10 and incubated with and without chaetocin for 24 h. A concentration of 200 nM chaetocin was chosen since it killed 100% of the A-498 ccRCC cell line while only killing 28% of CCD-18Co and 25% of CIK. The MTT assay can be used to determine the cytotoxicity of a certain drug on a single cell line by simply stating how many cells survived the treatment. However, the MTT assay cannot be used to distinguish between two different cell lines when incubated together. In order to determine how many non-cancerous and ccRCC cells survived the incubation along with chaetocin and CIK cells a FACS was used. Forward scatter (FSC) and side scatter (SSC) were used to identify and discriminate CIK cells, non-cancerous and ccRCC cells. The Hoechst 33258 staining solution was used in order to stain dead cells independent of being cancerous or non-cancerous to see how many survive the treatment. It was only possible to test one non-cancerous and one ccRCC cell line due to a lack of resources. Figure 5A 1.1 (Fig. 5) shows the population of CIK cells and A-498 ccRCC cells after 24 h of incubation in the absence of chaetocin. Both populations could be discriminated by their SSC and FSC. Figure 5A 1.2 (Fig. 5) shows the ratio of dead and living A-498 cells discriminated by Hoechst vs FSC. After the treatment 50.6% of the ccRCC cells were still alive while 45.9 % died and 3.5% could not be matched to either group. Figure 5A 2.1 and A 2.2 (Fig. 5) show an identical experimental set up but this time the cells were incubated in the presence of 200 nM chaetocin. After 24 h of incubation along with chaetocin 56.4% of the ccRCC cells died while only 41.5% stayed alive and 2.1% could not be matched to either group.

Bottom Line: The addition of distinct interleukins and antibodies results in the generation of CIK cells that are able to specifically target and destroy renal carcinoma cells.Although the CIK cells show a high grade of selective anti-ccRCC activity, this effect could not be improved by the addition of chaetocin.Abstract available from the publisher.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Integrated Oncology (CIO), University of Bonn, Bonn, Germany; Hochschule Bonn-Rhein-Sieg, Rheinbach, Germany.

ABSTRACT
We examined the cytotoxic effects of chaetocin on clear cell renal cell carcinoma (ccRCC) cells and the possibility to combine the effects of chaetocin with the effects of cytokine-induced killer cells (CIK) assayed by MTT assay and FACS analysis. Chaetocin is a thiodioxopiperazine produced by fungi belonging to the chaetomiaceae family. In 2007, it was first reported that chaetocin shows potent and selective ex vivo anti-cancer activity by inducing reactive oxygen species. CIK cells are generated from CD3+/CD56- T lymphocytes with double negative CD4-/CD8- phenotype that are isolated from human blood. The addition of distinct interleukins and antibodies results in the generation of CIK cells that are able to specifically target and destroy renal carcinoma cells. The results of this research state that the anti-ccRCC activity of chaetocin is weak and does not show a high grade of selectivity on clear cell renal cell carcinoma cells. Although the CIK cells show a high grade of selective anti-ccRCC activity, this effect could not be improved by the addition of chaetocin. So chaetocin seems to be no suitable agent for specific targeting ccRCC cells or for the combination therapy with CIK cells in renal cancer.

No MeSH data available.


Related in: MedlinePlus