Exosomal Carboxypeptidase E (CPE) and CPE-shRNA-Loaded Exosomes Regulate Metastatic Phenotype of Tumor Cells
Background: Exosomes promote tumor growth and metastasis through intercellular communication, although the mechanism remains elusive. Carboxypeptidase E (CPE) supports the progression of different cancers, including hepatocellular carcinoma (HCC). Here, we investigated whether CPE is the bioactive c...
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MDPI AG
2022-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/6/3113 |
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| author | Sangeetha Hareendran Bassam Albraidy Xuyu Yang Aiyi Liu Anne Breggia Clark C. Chen Y. Peng Loh |
| author_facet | Sangeetha Hareendran Bassam Albraidy Xuyu Yang Aiyi Liu Anne Breggia Clark C. Chen Y. Peng Loh |
| author_sort | Sangeetha Hareendran |
| collection | DOAJ |
| description | Background: Exosomes promote tumor growth and metastasis through intercellular communication, although the mechanism remains elusive. Carboxypeptidase E (CPE) supports the progression of different cancers, including hepatocellular carcinoma (HCC). Here, we investigated whether CPE is the bioactive cargo within exosomes, and whether it contributes to tumorigenesis, using HCC cell lines as a cancer model. Methods: Exosomes were isolated from supernatant media of cancer cells, or human sera. mRNA and protein expression were analyzed using PCR and Western blot. Low-metastatic HCC97L cells were incubated with exosomes derived from high-metastatic HCC97H cells. In other experiments, HCC97H cells were incubated with CPE-shRNA-loaded exosomes. Cell proliferation and invasion were assessed using MTT, colony formation, and matrigel invasion assays. Results: Exosomes released from cancer cells contain <i>CPE</i> mRNA and protein. <i>CPE</i> mRNA levels are enriched in exosomes secreted from high- versus low-metastastic cells, across various cancer types. In a pilot study, significantly higher <i>CPE</i> copy numbers were found in serum exosomes from cancer patients compared to healthy subjects. HCC97L cells, treated with exosomes derived from HCC97H cells, displayed enhanced proliferation and invasion; however, exosomes from HCC97H cells pre-treated with CPE-shRNA failed to promote proliferation. When HEK293T exosomes loaded with CPE-shRNA were incubated with HCC97H cells, the expression of CPE, Cyclin D1, a cell-cycle regulatory protein and <i>c-myc</i>, a proto-oncogene, were suppressed, resulting in the diminished proliferation of HCC97H cells. Conclusions: We identified CPE as an exosomal bioactive molecule driving the growth and invasion of low-metastatic HCC cells. CPE-shRNA loaded exosomes can inhibit malignant tumor cell proliferation via Cyclin D1 and c-MYC suppression. Thus, CPE is a key player in the exosome transmission of tumorigenesis, and the exosome-based delivery of CPE-shRNA offers a potential treatment for tumor progression. Notably, measuring CPE transcript levels in serum exosomes from cancer patients could have potential liquid biopsy applications. |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T19:42:03Z |
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| spelling | doaj.art-0fa108b3f600414ab031911f2a9c55b02023-11-24T01:32:45ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-03-01236311310.3390/ijms23063113Exosomal Carboxypeptidase E (CPE) and CPE-shRNA-Loaded Exosomes Regulate Metastatic Phenotype of Tumor CellsSangeetha Hareendran0Bassam Albraidy1Xuyu Yang2Aiyi Liu3Anne Breggia4Clark C. Chen5Y. Peng Loh6Section on Cellular Neurobiology, <i>Eunice Kennedy Shriver</i> National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USASection on Cellular Neurobiology, <i>Eunice Kennedy Shriver</i> National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USASection on Cellular Neurobiology, <i>Eunice Kennedy Shriver</i> National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USABiostatistics & Bioinformatics Branch, <i>Eunice Kennedy Shriver</i> National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USAMaine Medical Center BioBank, Portland, ME 04074, USADepartment of Neurosurgery, University of Minnesota Medical School, Minneapolis, MN 55455, USASection on Cellular Neurobiology, <i>Eunice Kennedy Shriver</i> National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USABackground: Exosomes promote tumor growth and metastasis through intercellular communication, although the mechanism remains elusive. Carboxypeptidase E (CPE) supports the progression of different cancers, including hepatocellular carcinoma (HCC). Here, we investigated whether CPE is the bioactive cargo within exosomes, and whether it contributes to tumorigenesis, using HCC cell lines as a cancer model. Methods: Exosomes were isolated from supernatant media of cancer cells, or human sera. mRNA and protein expression were analyzed using PCR and Western blot. Low-metastatic HCC97L cells were incubated with exosomes derived from high-metastatic HCC97H cells. In other experiments, HCC97H cells were incubated with CPE-shRNA-loaded exosomes. Cell proliferation and invasion were assessed using MTT, colony formation, and matrigel invasion assays. Results: Exosomes released from cancer cells contain <i>CPE</i> mRNA and protein. <i>CPE</i> mRNA levels are enriched in exosomes secreted from high- versus low-metastastic cells, across various cancer types. In a pilot study, significantly higher <i>CPE</i> copy numbers were found in serum exosomes from cancer patients compared to healthy subjects. HCC97L cells, treated with exosomes derived from HCC97H cells, displayed enhanced proliferation and invasion; however, exosomes from HCC97H cells pre-treated with CPE-shRNA failed to promote proliferation. When HEK293T exosomes loaded with CPE-shRNA were incubated with HCC97H cells, the expression of CPE, Cyclin D1, a cell-cycle regulatory protein and <i>c-myc</i>, a proto-oncogene, were suppressed, resulting in the diminished proliferation of HCC97H cells. Conclusions: We identified CPE as an exosomal bioactive molecule driving the growth and invasion of low-metastatic HCC cells. CPE-shRNA loaded exosomes can inhibit malignant tumor cell proliferation via Cyclin D1 and c-MYC suppression. Thus, CPE is a key player in the exosome transmission of tumorigenesis, and the exosome-based delivery of CPE-shRNA offers a potential treatment for tumor progression. Notably, measuring CPE transcript levels in serum exosomes from cancer patients could have potential liquid biopsy applications.https://www.mdpi.com/1422-0067/23/6/3113cancer proliferationhepatocellular carcinomametastasisengineered exosomesdiagnostic biomarkercancer therapy |
| spellingShingle | Sangeetha Hareendran Bassam Albraidy Xuyu Yang Aiyi Liu Anne Breggia Clark C. Chen Y. Peng Loh Exosomal Carboxypeptidase E (CPE) and CPE-shRNA-Loaded Exosomes Regulate Metastatic Phenotype of Tumor Cells International Journal of Molecular Sciences cancer proliferation hepatocellular carcinoma metastasis engineered exosomes diagnostic biomarker cancer therapy |
| title | Exosomal Carboxypeptidase E (CPE) and CPE-shRNA-Loaded Exosomes Regulate Metastatic Phenotype of Tumor Cells |
| title_full | Exosomal Carboxypeptidase E (CPE) and CPE-shRNA-Loaded Exosomes Regulate Metastatic Phenotype of Tumor Cells |
| title_fullStr | Exosomal Carboxypeptidase E (CPE) and CPE-shRNA-Loaded Exosomes Regulate Metastatic Phenotype of Tumor Cells |
| title_full_unstemmed | Exosomal Carboxypeptidase E (CPE) and CPE-shRNA-Loaded Exosomes Regulate Metastatic Phenotype of Tumor Cells |
| title_short | Exosomal Carboxypeptidase E (CPE) and CPE-shRNA-Loaded Exosomes Regulate Metastatic Phenotype of Tumor Cells |
| title_sort | exosomal carboxypeptidase e cpe and cpe shrna loaded exosomes regulate metastatic phenotype of tumor cells |
| topic | cancer proliferation hepatocellular carcinoma metastasis engineered exosomes diagnostic biomarker cancer therapy |
| url | https://www.mdpi.com/1422-0067/23/6/3113 |
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