CpG Site-Specific Methylation-Modulated Divergent Expression of PRSS3 Transcript Variants Facilitates Nongenetic Intratumor Heterogeneity in Human Hepatocellular Carcinoma
BackgroundHepatocellular carcinoma (HCC) is one of the most lethal human tumors with extensive intratumor heterogeneity (ITH). Serine protease 3 (PRSS3) is an indispensable member of the trypsin family and has been implicated in the pathogenesis of several malignancies, including HCC. However, the p...
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Frontiers Media S.A.
2022-04-01
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| Series: | Frontiers in Oncology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fonc.2022.831268/full |
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| author | Shuye Lin Hanli Xu Mengdi Pang Xiaomeng Zhou Xiaomeng Zhou Yuanming Pan Lishu Zhang Xin Guan Xiaoyue Wang Bonan Lin Rongmeng Tian Keqiang Chen Xiaochen Zhang Zijiang Yang Fengmin Ji Yingying Huang Wu Wei Wanghua Gong Jianke Ren Ji Ming Wang Mingzhou Guo Jiaqiang Huang Jiaqiang Huang Jiaqiang Huang |
| author_facet | Shuye Lin Hanli Xu Mengdi Pang Xiaomeng Zhou Xiaomeng Zhou Yuanming Pan Lishu Zhang Xin Guan Xiaoyue Wang Bonan Lin Rongmeng Tian Keqiang Chen Xiaochen Zhang Zijiang Yang Fengmin Ji Yingying Huang Wu Wei Wanghua Gong Jianke Ren Ji Ming Wang Mingzhou Guo Jiaqiang Huang Jiaqiang Huang Jiaqiang Huang |
| author_sort | Shuye Lin |
| collection | DOAJ |
| description | BackgroundHepatocellular carcinoma (HCC) is one of the most lethal human tumors with extensive intratumor heterogeneity (ITH). Serine protease 3 (PRSS3) is an indispensable member of the trypsin family and has been implicated in the pathogenesis of several malignancies, including HCC. However, the paradoxical effects of PRSS3 on carcinogenesis due to an unclear molecular basis impede the utilization of its biomarker potential. We hereby explored the contribution of PRSS3 transcripts to tumor functional heterogeneity by systematically dissecting the expression of four known splice variants of PRSS3 (PRSS3-SVs, V1~V4) and their functional relevance to HCC.MethodsThe expression and DNA methylation of PRSS3 transcripts and their associated clinical relevance in HCC were analyzed using several publicly available datasets and validated using qPCR-based assays. Functional experiments were performed in gain- and loss-of-function cell models, in which PRSS3 transcript constructs were separately transfected after deleting PRSS3 expression by CRISPR/Cas9 editing.ResultsPRSS3 was aberrantly differentially expressed toward bipolarity from very low (PRSS3Low) to very high (PRSS3High) expression across HCC cell lines and tissues. This was attributable to the disruption of PRSS3-SVs, in which PRSS3-V2 and/or PRSS3-V1 were dominant transcripts leading to PRSS3 expression, whereas PRSS3-V3 and -V4 were rarely or minimally expressed. The expression of PRSS3-V2 or -V1 was inversely associated with site-specific CpG methylation at the PRSS3 promoter region that distinguished HCC cells and tissues phenotypically between hypermethylated low-expression (mPRSS3-SVLow) and hypomethylated high-expression (umPRSS3-SVHigh) groups. PRSS3-SVs displayed distinct functions from oncogenic PRSS3-V2 to tumor-suppressive PRSS3-V1, -V3 or PRSS3-V4 in HCC cells. Clinically, aberrant expression of PRSS3-SVs was translated into divergent relevance in patients with HCC, in which significant epigenetic downregulation of PRSS3-V2 was seen in early HCC and was associated with favorable patient outcome.ConclusionsThese results provide the first evidence for the transcriptional and functional characterization of PRSS3 transcripts in HCC. Aberrant expression of divergent PRSS3-SVs disrupted by site-specific CpG methylation may integrate the effects of oncogenic PRSS3-V2 and tumor-suppressive PRSS3-V1, resulting in the molecular diversity and functional plasticity of PRSS3 in HCC. Dysregulated expression of PRSS3-V2 by site-specific CpG methylation may have potential diagnostic value for patients with early HCC. |
| first_indexed | 2024-12-17T06:56:35Z |
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| series | Frontiers in Oncology |
| spelling | doaj.art-3a0aaf074ab648fa893225acf5efb4122022-12-21T21:59:25ZengFrontiers Media S.A.Frontiers in Oncology2234-943X2022-04-011210.3389/fonc.2022.831268831268CpG Site-Specific Methylation-Modulated Divergent Expression of PRSS3 Transcript Variants Facilitates Nongenetic Intratumor Heterogeneity in Human Hepatocellular CarcinomaShuye Lin0Hanli Xu1Mengdi Pang2Xiaomeng Zhou3Xiaomeng Zhou4Yuanming Pan5Lishu Zhang6Xin Guan7Xiaoyue Wang8Bonan Lin9Rongmeng Tian10Keqiang Chen11Xiaochen Zhang12Zijiang Yang13Fengmin Ji14Yingying Huang15Wu Wei16Wanghua Gong17Jianke Ren18Ji Ming Wang19Mingzhou Guo20Jiaqiang Huang21Jiaqiang Huang22Jiaqiang Huang23Cancer Research Center, Beijing Chest Hospital, Capital Medical University, Beijing Tuberculosis and Thoracic Tumor Institute, Beijing, ChinaCollege of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, ChinaCollege of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, ChinaCollege of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, ChinaDepartment of Gastroenterology and Hepatology, Chinese People’s Liberation Army of China (PLA) General Hospital, Beijing, ChinaCancer Research Center, Beijing Chest Hospital, Capital Medical University, Beijing Tuberculosis and Thoracic Tumor Institute, Beijing, ChinaCollege of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, ChinaCollege of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, ChinaCollege of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, ChinaCollege of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, ChinaCollege of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, ChinaLaboratory of Cancer Immunometabolism, Center for Cancer Research, National Cancer Institute, Frederick, MD, United StatesCollege of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, ChinaCollege of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, ChinaCollege of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, ChinaChinese Academy of Sciences (CAS) Key Laboratory of Computational Biology, Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, ChinaChinese Academy of Sciences (CAS) Key Laboratory of Computational Biology, Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, ChinaBasic Research Program, Leidos Biomedical Research, Inc., Frederick, MD, United StatesChinese Academy of Sciences (CAS) Key Laboratory of Computational Biology, Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, ChinaLaboratory of Cancer Immunometabolism, Center for Cancer Research, National Cancer Institute, Frederick, MD, United StatesDepartment of Gastroenterology and Hepatology, Chinese People’s Liberation Army of China (PLA) General Hospital, Beijing, ChinaCancer Research Center, Beijing Chest Hospital, Capital Medical University, Beijing Tuberculosis and Thoracic Tumor Institute, Beijing, ChinaCollege of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing, ChinaLaboratory of Cancer Immunometabolism, Center for Cancer Research, National Cancer Institute, Frederick, MD, United StatesBackgroundHepatocellular carcinoma (HCC) is one of the most lethal human tumors with extensive intratumor heterogeneity (ITH). Serine protease 3 (PRSS3) is an indispensable member of the trypsin family and has been implicated in the pathogenesis of several malignancies, including HCC. However, the paradoxical effects of PRSS3 on carcinogenesis due to an unclear molecular basis impede the utilization of its biomarker potential. We hereby explored the contribution of PRSS3 transcripts to tumor functional heterogeneity by systematically dissecting the expression of four known splice variants of PRSS3 (PRSS3-SVs, V1~V4) and their functional relevance to HCC.MethodsThe expression and DNA methylation of PRSS3 transcripts and their associated clinical relevance in HCC were analyzed using several publicly available datasets and validated using qPCR-based assays. Functional experiments were performed in gain- and loss-of-function cell models, in which PRSS3 transcript constructs were separately transfected after deleting PRSS3 expression by CRISPR/Cas9 editing.ResultsPRSS3 was aberrantly differentially expressed toward bipolarity from very low (PRSS3Low) to very high (PRSS3High) expression across HCC cell lines and tissues. This was attributable to the disruption of PRSS3-SVs, in which PRSS3-V2 and/or PRSS3-V1 were dominant transcripts leading to PRSS3 expression, whereas PRSS3-V3 and -V4 were rarely or minimally expressed. The expression of PRSS3-V2 or -V1 was inversely associated with site-specific CpG methylation at the PRSS3 promoter region that distinguished HCC cells and tissues phenotypically between hypermethylated low-expression (mPRSS3-SVLow) and hypomethylated high-expression (umPRSS3-SVHigh) groups. PRSS3-SVs displayed distinct functions from oncogenic PRSS3-V2 to tumor-suppressive PRSS3-V1, -V3 or PRSS3-V4 in HCC cells. Clinically, aberrant expression of PRSS3-SVs was translated into divergent relevance in patients with HCC, in which significant epigenetic downregulation of PRSS3-V2 was seen in early HCC and was associated with favorable patient outcome.ConclusionsThese results provide the first evidence for the transcriptional and functional characterization of PRSS3 transcripts in HCC. Aberrant expression of divergent PRSS3-SVs disrupted by site-specific CpG methylation may integrate the effects of oncogenic PRSS3-V2 and tumor-suppressive PRSS3-V1, resulting in the molecular diversity and functional plasticity of PRSS3 in HCC. Dysregulated expression of PRSS3-V2 by site-specific CpG methylation may have potential diagnostic value for patients with early HCC.https://www.frontiersin.org/articles/10.3389/fonc.2022.831268/fullliver cancerhepatocellular carcinomaPRSS3transcript variantintratumor heterogeneityCpG methylation |
| spellingShingle | Shuye Lin Hanli Xu Mengdi Pang Xiaomeng Zhou Xiaomeng Zhou Yuanming Pan Lishu Zhang Xin Guan Xiaoyue Wang Bonan Lin Rongmeng Tian Keqiang Chen Xiaochen Zhang Zijiang Yang Fengmin Ji Yingying Huang Wu Wei Wanghua Gong Jianke Ren Ji Ming Wang Mingzhou Guo Jiaqiang Huang Jiaqiang Huang Jiaqiang Huang CpG Site-Specific Methylation-Modulated Divergent Expression of PRSS3 Transcript Variants Facilitates Nongenetic Intratumor Heterogeneity in Human Hepatocellular Carcinoma Frontiers in Oncology liver cancer hepatocellular carcinoma PRSS3 transcript variant intratumor heterogeneity CpG methylation |
| title | CpG Site-Specific Methylation-Modulated Divergent Expression of PRSS3 Transcript Variants Facilitates Nongenetic Intratumor Heterogeneity in Human Hepatocellular Carcinoma |
| title_full | CpG Site-Specific Methylation-Modulated Divergent Expression of PRSS3 Transcript Variants Facilitates Nongenetic Intratumor Heterogeneity in Human Hepatocellular Carcinoma |
| title_fullStr | CpG Site-Specific Methylation-Modulated Divergent Expression of PRSS3 Transcript Variants Facilitates Nongenetic Intratumor Heterogeneity in Human Hepatocellular Carcinoma |
| title_full_unstemmed | CpG Site-Specific Methylation-Modulated Divergent Expression of PRSS3 Transcript Variants Facilitates Nongenetic Intratumor Heterogeneity in Human Hepatocellular Carcinoma |
| title_short | CpG Site-Specific Methylation-Modulated Divergent Expression of PRSS3 Transcript Variants Facilitates Nongenetic Intratumor Heterogeneity in Human Hepatocellular Carcinoma |
| title_sort | cpg site specific methylation modulated divergent expression of prss3 transcript variants facilitates nongenetic intratumor heterogeneity in human hepatocellular carcinoma |
| topic | liver cancer hepatocellular carcinoma PRSS3 transcript variant intratumor heterogeneity CpG methylation |
| url | https://www.frontiersin.org/articles/10.3389/fonc.2022.831268/full |
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