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Oncolytic Activity of a Recombinant Measles Virus, Blind to Signaling Lymphocyte Activation Molecule, Against Colorectal Cancer Cells.

Amagai Y, Fujiyuki T, Yoneda M, Shoji K, Furukawa Y, Sato H, Kai C - Sci Rep (2016)

Bottom Line: Oncolytic virotherapy is a distinctive antitumor therapy based on the cancer-cell-specific infectivity and killing activity of viruses, which exert a considerable antitumor effect with only a few treatments.Tumour progression in xenograft models was also abrogated by the virus, and the infection of cancer cells in vivo by the virus was demonstrated with both flow cytometry and a histological analysis.Therefore, rMV-SLAMblind is considered a novel therapeutic agent for colorectal cancers, including those resistant to molecular-targeted therapies.

View Article: PubMed Central - PubMed

Affiliation: Laboratory Animal Research Center, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.

ABSTRACT
Oncolytic virotherapy is a distinctive antitumor therapy based on the cancer-cell-specific infectivity and killing activity of viruses, which exert a considerable antitumor effect with only a few treatments. Because colorectal cancer cells often acquire resistance to the molecular-targeted therapies and alternative treatments are called for, in this study, we evaluated the oncolytic activity against colorectal cancer cells of a recombinant measles virus (rMV-SLAMblind), which is blind to signaling lymphocytic activation molecule (SLAM) and infects target cells via nectin-4/poliovirus receptor-related 4 protein. We examined 10 cell lines including 8 cell lines that were resistant to epidermal-growth-factor-receptor (EGFR) targeted therapy. rMV-SLAMblind infected and lysed the nectin-4-positive cell lines dependently on nectin-4 expression, in spite of mutation in EGFR cascade. Tumour progression in xenograft models was also abrogated by the virus, and the infection of cancer cells in vivo by the virus was demonstrated with both flow cytometry and a histological analysis. Therefore, rMV-SLAMblind is considered a novel therapeutic agent for colorectal cancers, including those resistant to molecular-targeted therapies.

No MeSH data available.


Related in: MedlinePlus

Expression of MV receptors on colorectal cancer cells.(a) Flow-cytometric analysis of cell-surface proteins that are associated with MV entry. A total number of 106 cells were stained with each primary and secondary Ab (black line). For nectin-4 detection, mAb (clone N4.61) was used in this experiment. The grey histogram indicates the IgG control for each cell line. Shown are representative data of three independent experiments. (b) RT-PCR analysis of each colorectal cancer cell line. Representative data of the electrophoresis (left) as well as relative expression level of nectin-4 based on the qRT-PCR analysis (right) is presented. In the qRT-PCR analysis, expression level of nectin-4 was normalized to GAPDH, and the average ± SD of three independent experiments is indicated.
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f1: Expression of MV receptors on colorectal cancer cells.(a) Flow-cytometric analysis of cell-surface proteins that are associated with MV entry. A total number of 106 cells were stained with each primary and secondary Ab (black line). For nectin-4 detection, mAb (clone N4.61) was used in this experiment. The grey histogram indicates the IgG control for each cell line. Shown are representative data of three independent experiments. (b) RT-PCR analysis of each colorectal cancer cell line. Representative data of the electrophoresis (left) as well as relative expression level of nectin-4 based on the qRT-PCR analysis (right) is presented. In the qRT-PCR analysis, expression level of nectin-4 was normalized to GAPDH, and the average ± SD of three independent experiments is indicated.

Mentions: A flow-cytometric analysis was conducted to examine the expression of nectin-4 in colorectal cancer cell lines. Among the 10 cell lines examined (CaCo-2, DLD1, HT29, LS174T, SW48, SW948, HCT116, LoVo, RKO, and SW480), the CaCo-2, DLD1, HT29, LS174T, SW48, and SW948 cell lines expressed nectin-4, whereas the others did not (Fig. 1a, Table 1). Among these, nectin-4 expression in SW48 cells was heterogeneous, of which approximately 15% cells only express nectin-4 on the cell surface (Fig. 1a). The wild-type MV strains, including the HL strain, use SLAM as their receptor, whereas MV vaccine strains use CD461420, which is a recognition molecule expressed ubiquitously in human nucleated cells. We also analysed the expression of these receptors and observed that all the cell lines tested were negative for SLAM and positive for CD46 (Fig. 1a). To ascertain the expression of nectin-4 at the messenger RNA (mRNA) level, reverse transcription and polymerase chain reaction (RT-PCR) were performed. Higher expression of nectin-4 mRNA was observed in the cells that were positive for nectin-4 in the flow-cytometric analysis than in those that were nectin-4-negative on flow cytometry (Fig. 1a,b). Regarding SW48 cells, nectin-4 mRNA expression was as high as other nectin-4-positive cells in spite of their heterogeneous nectin-4 expression (Fig. 1b).


Oncolytic Activity of a Recombinant Measles Virus, Blind to Signaling Lymphocyte Activation Molecule, Against Colorectal Cancer Cells.

Amagai Y, Fujiyuki T, Yoneda M, Shoji K, Furukawa Y, Sato H, Kai C - Sci Rep (2016)

Expression of MV receptors on colorectal cancer cells.(a) Flow-cytometric analysis of cell-surface proteins that are associated with MV entry. A total number of 106 cells were stained with each primary and secondary Ab (black line). For nectin-4 detection, mAb (clone N4.61) was used in this experiment. The grey histogram indicates the IgG control for each cell line. Shown are representative data of three independent experiments. (b) RT-PCR analysis of each colorectal cancer cell line. Representative data of the electrophoresis (left) as well as relative expression level of nectin-4 based on the qRT-PCR analysis (right) is presented. In the qRT-PCR analysis, expression level of nectin-4 was normalized to GAPDH, and the average ± SD of three independent experiments is indicated.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835780&req=5

f1: Expression of MV receptors on colorectal cancer cells.(a) Flow-cytometric analysis of cell-surface proteins that are associated with MV entry. A total number of 106 cells were stained with each primary and secondary Ab (black line). For nectin-4 detection, mAb (clone N4.61) was used in this experiment. The grey histogram indicates the IgG control for each cell line. Shown are representative data of three independent experiments. (b) RT-PCR analysis of each colorectal cancer cell line. Representative data of the electrophoresis (left) as well as relative expression level of nectin-4 based on the qRT-PCR analysis (right) is presented. In the qRT-PCR analysis, expression level of nectin-4 was normalized to GAPDH, and the average ± SD of three independent experiments is indicated.
Mentions: A flow-cytometric analysis was conducted to examine the expression of nectin-4 in colorectal cancer cell lines. Among the 10 cell lines examined (CaCo-2, DLD1, HT29, LS174T, SW48, SW948, HCT116, LoVo, RKO, and SW480), the CaCo-2, DLD1, HT29, LS174T, SW48, and SW948 cell lines expressed nectin-4, whereas the others did not (Fig. 1a, Table 1). Among these, nectin-4 expression in SW48 cells was heterogeneous, of which approximately 15% cells only express nectin-4 on the cell surface (Fig. 1a). The wild-type MV strains, including the HL strain, use SLAM as their receptor, whereas MV vaccine strains use CD461420, which is a recognition molecule expressed ubiquitously in human nucleated cells. We also analysed the expression of these receptors and observed that all the cell lines tested were negative for SLAM and positive for CD46 (Fig. 1a). To ascertain the expression of nectin-4 at the messenger RNA (mRNA) level, reverse transcription and polymerase chain reaction (RT-PCR) were performed. Higher expression of nectin-4 mRNA was observed in the cells that were positive for nectin-4 in the flow-cytometric analysis than in those that were nectin-4-negative on flow cytometry (Fig. 1a,b). Regarding SW48 cells, nectin-4 mRNA expression was as high as other nectin-4-positive cells in spite of their heterogeneous nectin-4 expression (Fig. 1b).

Bottom Line: Oncolytic virotherapy is a distinctive antitumor therapy based on the cancer-cell-specific infectivity and killing activity of viruses, which exert a considerable antitumor effect with only a few treatments.Tumour progression in xenograft models was also abrogated by the virus, and the infection of cancer cells in vivo by the virus was demonstrated with both flow cytometry and a histological analysis.Therefore, rMV-SLAMblind is considered a novel therapeutic agent for colorectal cancers, including those resistant to molecular-targeted therapies.

View Article: PubMed Central - PubMed

Affiliation: Laboratory Animal Research Center, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.

ABSTRACT
Oncolytic virotherapy is a distinctive antitumor therapy based on the cancer-cell-specific infectivity and killing activity of viruses, which exert a considerable antitumor effect with only a few treatments. Because colorectal cancer cells often acquire resistance to the molecular-targeted therapies and alternative treatments are called for, in this study, we evaluated the oncolytic activity against colorectal cancer cells of a recombinant measles virus (rMV-SLAMblind), which is blind to signaling lymphocytic activation molecule (SLAM) and infects target cells via nectin-4/poliovirus receptor-related 4 protein. We examined 10 cell lines including 8 cell lines that were resistant to epidermal-growth-factor-receptor (EGFR) targeted therapy. rMV-SLAMblind infected and lysed the nectin-4-positive cell lines dependently on nectin-4 expression, in spite of mutation in EGFR cascade. Tumour progression in xenograft models was also abrogated by the virus, and the infection of cancer cells in vivo by the virus was demonstrated with both flow cytometry and a histological analysis. Therefore, rMV-SLAMblind is considered a novel therapeutic agent for colorectal cancers, including those resistant to molecular-targeted therapies.

No MeSH data available.


Related in: MedlinePlus