Pengbo Cao1, Aiqing Yang2, Rui Wang3, Xia Xia1, Yun Zhai1, Yuanfeng Li1, Fei Yang1, Ying Cui4, Weimin Xie4, Ying Liu5, Taotao Liu6, Weihua Jia7, Zhengwen Jiang8, Zhuo Li9, Yuqing Han1, Chengming Gao1, Qingfeng Song10, Bobo Xie1, Luo Zhang11, Hongxing Zhang1, Jinxu Zhang1, Xizhong Shen6, Yunfei Yuan7, Feng Yu8, Ying Wang8, Jing Xu8, Yilong Ma10, Zengnan Mo12, Wuzhong Yu13, Fuchu He14, Gangqiao Zhou15. 1. State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing, P. R. China; National Engineering Research Center for Protein Drugs, Beijing, P. R. China; National Center for Protein Sciences at Beijing, Beijing, P. R. China. 2. School of Life Sciences, Tsinghua University, Beijing, P. R. China. 3. State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing, P. R. China; National Engineering Research Center for Protein Drugs, Beijing, P. R. China; National Center for Protein Sciences at Beijing, Beijing, P. R. China; Clinical Medical Institute of General Hospital of Xinjiang Military Area, PLA, Urumqi, Xinjiang, P. R. China. 4. Affiliated Cancer Hospital of Guangxi Medical University, Nanning, Guangxi, P. R. China. 5. National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences; School of Basic Medicine, Beijing, P. R. China. 6. Department of Gastroenterology and Hepatology, Zhongshan Hospital, Fudan University, Shanghai, P. R. China. 7. Department of Experimental Research, Cancer Center, Sun Yat-sen University, Guangzhou, Guangdong, P. R. China. 8. Center for Genetics and Genomics Analysis, Genesky Biotechnologies Inc., Shanghai, P. R. China. 9. Department of Infectious Diseases, Affiliated Youan Hospital, Capital University of Medical Science, Beijing, P. R. China. 10. Interventional Radiology Department of Affiliated Tumor Hospital of Guangxi Medical University, Nanning, Guangxi, P. R. China. 11. State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing, P. R. China. 12. Department of Urology and Nephrology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, P. R. China. 13. Clinical Medical Institute of General Hospital of Xinjiang Military Area, PLA, Urumqi, Xinjiang, P. R. China. 14. State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing, P. R. China; National Engineering Research Center for Protein Drugs, Beijing, P. R. China; National Center for Protein Sciences at Beijing, Beijing, P. R. China; School of Life Sciences, Tsinghua University, Beijing, P. R. China. Electronic address: hefc@nic.bmi.ac.cn. 15. State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing, P. R. China; National Engineering Research Center for Protein Drugs, Beijing, P. R. China; National Center for Protein Sciences at Beijing, Beijing, P. R. China. Electronic address: zhougq114@126.com.
Abstract
BACKGROUND & AIMS: Single nucleotide polymorphisms could affect risk for hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). We performed a germline copy number variation (CNV)-based genome-wide association study (GWAS) in populations of Chinese ancestry to search for germline CNVs that increase risk of HCC. METHODS: We conducted a CNV-based GWAS of 1583 HCC cases (persons with chronic HBV infection and HCC) and 1540 controls (persons with chronic HBV infection without HCC) in Chinese populations. Identified candidates were expressed in L-02, HepG2, or TP53-/- or wild-type HCT116 cells, and knocked down with short hairpin RNAs in HepG2, Bel-7402, and SMMC-7721 cells; proliferation, colony formation, and apoptosis were measured. Formation of xenograft tumors from cell lines was monitored in nude mice. Subcellular localization of ribosome proteins and levels or activity of p53 were investigated by co-immunoprecipitation, immunofluorescence, and immunoblot analyses. Levels of small nucleolar RNA H/ACA box 18-like 5 (SNORA18L5) were quantified by quantitative reverse transcription polymerase chain reaction. RESULTS: We identified a low-frequency duplication at chromosome 15q13.3 strongly associated with risk of HBV-related HCC (overall P = 3.17 × 10-8; odds ratio, 12.02). Copy numbers of the 15q13.3 duplication correlated with the expression of SNORA18L5 in liver tissues. Overexpression of SNORA18L5 increased HCC cell proliferation and growth of xenograft tumors in mice; knockdown reduced HCC proliferation and tumor growth. SNORA18L5 overexpression in HepG2 and SMMC-7721 cells inhibited p53-dependent cell cycle arrest and apoptosis. Overexpression of SNORA18L5 led to hyperactive ribosome biogenesis, increasing levels of mature 18S and 28S ribosomal RNAs and causing the ribosomal proteins RPL5 and RPL11 to stay in the nucleolus, which kept them from binding to MDM2. This resulted in increased MDM2-mediated ubiquitination and degradation of p53. Levels of SNORA18L5 were increased in HCC tissues compared with nontumor liver tissues and associated with shorter survival times of patients. CONCLUSIONS: In a CNV-based GWAS, we associated duplication at 15q13.3 with increased risk of HBV-related HCC. We found SNORA18L5 at this location to promote HCC cell proliferation and tumor growth in mice. SNORA18L5 increases ribosome biogenesis, facilitates ribosomal RNA maturation, and alters localization of RPL5 and RPL11, allowing for increased MDM2-mediated proteolysis of p53 and cell cycle arrest.
BACKGROUND & AIMS: Single nucleotide polymorphisms could affect risk for hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). We performed a germline copy number variation (CNV)-based genome-wide association study (GWAS) in populations of Chinese ancestry to search for germline CNVs that increase risk of HCC. METHODS: We conducted a CNV-based GWAS of 1583 HCC cases (persons with chronic HBV infection and HCC) and 1540 controls (persons with chronic HBV infection without HCC) in Chinese populations. Identified candidates were expressed in L-02, HepG2, or TP53-/- or wild-type HCT116 cells, and knocked down with short hairpin RNAs in HepG2, Bel-7402, and SMMC-7721 cells; proliferation, colony formation, and apoptosis were measured. Formation of xenograft tumors from cell lines was monitored in nude mice. Subcellular localization of ribosome proteins and levels or activity of p53 were investigated by co-immunoprecipitation, immunofluorescence, and immunoblot analyses. Levels of small nucleolar RNA H/ACA box 18-like 5 (SNORA18L5) were quantified by quantitative reverse transcription polymerase chain reaction. RESULTS: We identified a low-frequency duplication at chromosome 15q13.3 strongly associated with risk of HBV-related HCC (overall P = 3.17 × 10-8; odds ratio, 12.02). Copy numbers of the 15q13.3 duplication correlated with the expression of SNORA18L5 in liver tissues. Overexpression of SNORA18L5 increased HCC cell proliferation and growth of xenograft tumors in mice; knockdown reduced HCC proliferation and tumor growth. SNORA18L5 overexpression in HepG2 and SMMC-7721 cells inhibited p53-dependent cell cycle arrest and apoptosis. Overexpression of SNORA18L5 led to hyperactive ribosome biogenesis, increasing levels of mature 18S and 28S ribosomal RNAs and causing the ribosomal proteins RPL5 and RPL11 to stay in the nucleolus, which kept them from binding to MDM2. This resulted in increased MDM2-mediated ubiquitination and degradation of p53. Levels of SNORA18L5 were increased in HCC tissues compared with nontumor liver tissues and associated with shorter survival times of patients. CONCLUSIONS: In a CNV-based GWAS, we associated duplication at 15q13.3 with increased risk of HBV-related HCC. We found SNORA18L5 at this location to promote HCC cell proliferation and tumor growth in mice. SNORA18L5 increases ribosome biogenesis, facilitates ribosomal RNA maturation, and alters localization of RPL5 and RPL11, allowing for increased MDM2-mediated proteolysis of p53 and cell cycle arrest.
Authors: Gilbert S Omenn; Lydie Lane; Christopher M Overall; Fernando J Corrales; Jochen M Schwenk; Young-Ki Paik; Jennifer E Van Eyk; Siqi Liu; Stephen Pennington; Michael P Snyder; Mark S Baker; Eric W Deutsch Journal: J Proteome Res Date: 2019-09-13 Impact factor: 4.466
Authors: Fathelrahman M Hassan; Afnan A Alsultan; Faisal Alzahrani; Waleed H Albuali; Dalal K Bubshait; Elfadil M Abass; Mudathir A Elbasheer; Abdulmohsen A Alkhanbashi Journal: Saudi Med J Date: 2021-09 Impact factor: 1.422