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Genetic Mutation and Exosome Signature of Human Papilloma Virus Associated Oropharyngeal Cancer

View Article: PubMed Central - PubMed

ABSTRACT

Human papilloma virus-16 (HPV-16) associated oropharyngeal cancer (HPVOPC) is increasing alarmingly in the United States. We performed whole genome sequencing of a 44 year old, male HPVOPC subject diagnosed with moderately differentiated tonsillar carcinoma. We identified new somatic mutation in MUC16 (A.k.a. CA-125), MUC12, MUC4, MUC6, MUC2, SIRPA, HLA-DRB1, HLA-A and HLA-B molecules. Increased protein expression of MUC16, SIRPA and decreased expression of HLA-DRB1 was further demonstrated in this HPVOPC subject and an additional set of 15 HPVOPC cases. Copy number gain (3 copies) was also observed for MUC2, MUC4, MUC6 and SIRPA. Enhanced expression of MUC16, SIRPA and HPV-16-E7 protein was detectable in the circulating exosomes of numerous HPVOPC subjects. Treatment of non-tumorigenic mammary epithelial cells with exosomes derived from aggressive HPVOPC cells harboring MUC16, SIRPA and HPV-16-E7 proteins augmented invasion and induced epithelial to mesenchymal transition (EMT) accompanied by an increased expression ratio of the EMT markers Vimentin/E-cadherin. Exosome based screening of key HPVOPC associated molecules could be beneficial for early cancer diagnosis, monitoring and surveillance.

No MeSH data available.


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Impact of exosomes treatment on EMT and invasion.(a) Confirmation of the exosome uptake (yellow arrowheads, upper panel) by the HMLE cells. Increased invasion (p = 0.0001) and induction of EMT phenotype (arrowheads) in the exosome treated HMLE cells. (b) Confirmation of the exosome uptake (red arrowheads, upper panel) by the MCF-10A cells. Enhanced invasion (p = 0.0001) and modulation of EMT phenotype (arrowheads) in the exosome treated MCF-10A cells. (c) Detection of increased Vimentin/E-cadharin expression ratio (panel 1 and 2) accompanied by enhanced expression of SIRPA and MUC16 (panel 3 and 4) in the cells treated with UM-SCC-104 cells’ secreted exosomes. Detection of HPV-16-E7 protein (panel 5) in the exosome treated cells (c). Actin was used as a loading control. Magnification X 200 (a,b). Control: cells treated with PBS; Treated: cells treated with UM-SCC-104 cells’ secreted exosomes.
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f5: Impact of exosomes treatment on EMT and invasion.(a) Confirmation of the exosome uptake (yellow arrowheads, upper panel) by the HMLE cells. Increased invasion (p = 0.0001) and induction of EMT phenotype (arrowheads) in the exosome treated HMLE cells. (b) Confirmation of the exosome uptake (red arrowheads, upper panel) by the MCF-10A cells. Enhanced invasion (p = 0.0001) and modulation of EMT phenotype (arrowheads) in the exosome treated MCF-10A cells. (c) Detection of increased Vimentin/E-cadharin expression ratio (panel 1 and 2) accompanied by enhanced expression of SIRPA and MUC16 (panel 3 and 4) in the cells treated with UM-SCC-104 cells’ secreted exosomes. Detection of HPV-16-E7 protein (panel 5) in the exosome treated cells (c). Actin was used as a loading control. Magnification X 200 (a,b). Control: cells treated with PBS; Treated: cells treated with UM-SCC-104 cells’ secreted exosomes.

Mentions: The UM-SCC-104 cell line used in this study was generated from a recurrent HPVOPC patient30 and the secreted exosomes from these cells were found to harbor HPV-16-E7, MUC16 and SIRPA proteins (Fig. 4d). To determine the impact of these exosomes on influencing cell growth, we co-cultured UM-SCC-104 secreted exosomes with a couple of non-tumorigenic and HPV-16− human mammary epithelial cell lines for 1 week and measured epithelial to mesenchymal transition (EMT) changes and invasion potential of these epithelial cells. First, we confirmed the entry of the exosomes in these cells (Fig. 5a,b arrowheads). We observed an EMT phenotype (arrowheads) as determined by cell morphology3132 accompanied by an increased invasion (p = 0.0001) of both the cell lines following the exosomes treatment (Fig. 5a,b). In addition, an increased expression ratio of the EMT markers Vimentin/E-cadherin was observed in these exosome treated cells accompanied by an enhanced expression of SIRPA and MUC16 (Fig. 5c). Notably, these mammary epithelial cells were also found to harbor HPV-16-E7 protein following the exosome treatment (Fig. 5c).


Genetic Mutation and Exosome Signature of Human Papilloma Virus Associated Oropharyngeal Cancer
Impact of exosomes treatment on EMT and invasion.(a) Confirmation of the exosome uptake (yellow arrowheads, upper panel) by the HMLE cells. Increased invasion (p = 0.0001) and induction of EMT phenotype (arrowheads) in the exosome treated HMLE cells. (b) Confirmation of the exosome uptake (red arrowheads, upper panel) by the MCF-10A cells. Enhanced invasion (p = 0.0001) and modulation of EMT phenotype (arrowheads) in the exosome treated MCF-10A cells. (c) Detection of increased Vimentin/E-cadharin expression ratio (panel 1 and 2) accompanied by enhanced expression of SIRPA and MUC16 (panel 3 and 4) in the cells treated with UM-SCC-104 cells’ secreted exosomes. Detection of HPV-16-E7 protein (panel 5) in the exosome treated cells (c). Actin was used as a loading control. Magnification X 200 (a,b). Control: cells treated with PBS; Treated: cells treated with UM-SCC-104 cells’ secreted exosomes.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5382706&req=5

f5: Impact of exosomes treatment on EMT and invasion.(a) Confirmation of the exosome uptake (yellow arrowheads, upper panel) by the HMLE cells. Increased invasion (p = 0.0001) and induction of EMT phenotype (arrowheads) in the exosome treated HMLE cells. (b) Confirmation of the exosome uptake (red arrowheads, upper panel) by the MCF-10A cells. Enhanced invasion (p = 0.0001) and modulation of EMT phenotype (arrowheads) in the exosome treated MCF-10A cells. (c) Detection of increased Vimentin/E-cadharin expression ratio (panel 1 and 2) accompanied by enhanced expression of SIRPA and MUC16 (panel 3 and 4) in the cells treated with UM-SCC-104 cells’ secreted exosomes. Detection of HPV-16-E7 protein (panel 5) in the exosome treated cells (c). Actin was used as a loading control. Magnification X 200 (a,b). Control: cells treated with PBS; Treated: cells treated with UM-SCC-104 cells’ secreted exosomes.
Mentions: The UM-SCC-104 cell line used in this study was generated from a recurrent HPVOPC patient30 and the secreted exosomes from these cells were found to harbor HPV-16-E7, MUC16 and SIRPA proteins (Fig. 4d). To determine the impact of these exosomes on influencing cell growth, we co-cultured UM-SCC-104 secreted exosomes with a couple of non-tumorigenic and HPV-16− human mammary epithelial cell lines for 1 week and measured epithelial to mesenchymal transition (EMT) changes and invasion potential of these epithelial cells. First, we confirmed the entry of the exosomes in these cells (Fig. 5a,b arrowheads). We observed an EMT phenotype (arrowheads) as determined by cell morphology3132 accompanied by an increased invasion (p = 0.0001) of both the cell lines following the exosomes treatment (Fig. 5a,b). In addition, an increased expression ratio of the EMT markers Vimentin/E-cadherin was observed in these exosome treated cells accompanied by an enhanced expression of SIRPA and MUC16 (Fig. 5c). Notably, these mammary epithelial cells were also found to harbor HPV-16-E7 protein following the exosome treatment (Fig. 5c).

View Article: PubMed Central - PubMed

ABSTRACT

Human papilloma virus-16 (HPV-16) associated oropharyngeal cancer (HPVOPC) is increasing alarmingly in the United States. We performed whole genome sequencing of a 44 year old, male HPVOPC subject diagnosed with moderately differentiated tonsillar carcinoma. We identified new somatic mutation in MUC16 (A.k.a. CA-125), MUC12, MUC4, MUC6, MUC2, SIRPA, HLA-DRB1, HLA-A and HLA-B molecules. Increased protein expression of MUC16, SIRPA and decreased expression of HLA-DRB1 was further demonstrated in this HPVOPC subject and an additional set of 15 HPVOPC cases. Copy number gain (3 copies) was also observed for MUC2, MUC4, MUC6 and SIRPA. Enhanced expression of MUC16, SIRPA and HPV-16-E7 protein was detectable in the circulating exosomes of numerous HPVOPC subjects. Treatment of non-tumorigenic mammary epithelial cells with exosomes derived from aggressive HPVOPC cells harboring MUC16, SIRPA and HPV-16-E7 proteins augmented invasion and induced epithelial to mesenchymal transition (EMT) accompanied by an increased expression ratio of the EMT markers Vimentin/E-cadherin. Exosome based screening of key HPVOPC associated molecules could be beneficial for early cancer diagnosis, monitoring and surveillance.

No MeSH data available.


Related in: MedlinePlus