PARProceedings of Anticancer Research2208-35452208-3553Bio-Byword Scientific Publishing Pty. Ltd.10.26689/par.v6i3.4011ArticleInhibition of RNaseH2A Induces Fas-Regulated Programmed Cell Death in Hepatoma Cellshttps://artdesignp.com/journal/PAR/6/3/10.26689/par.v6i3.4011ChangXiaowei,CaiYu,YanPu,LiWujun2022632022-05-30Objective: To screen clinically significant potential drug targets in liver cancer and to study the function and potential molecular mechanisms of this target protein in the development of liver cancer. Methods: By using the clinical database GEPIA to find genes that are differentially expressed in liver cancer compared to normal tissues, we further screened the genes that are highly expressed in hepatocellular carcinoma and have clinical prognostic relevance. Heat maps were used to sort these genes according to their clinical prognostic relevance, so as to screen for the target gene of interest. The characteristics of target gene expression and clinical prognosis in hepatocellular carcinoma were studied. The target gene was knocked down through siRNA, and cell proliferation experiments and apoptosis experiments were used to verify the importance of the target gene in the occurrence and development of liver cancer. Finally, we elucidated the potential molecular mechanism of the target gene’s function in liver cancer based on the mutual regulatory relationship between the target gene and key apoptosis genes. Results: 1482 genes were significantly underexpressed in liver cancer, and 725 genes were significantly overexpressed in liver cancer, of which RNaseH2A was significantly overexpressed in liver cancer and had a significant clinical prognosis. Knockdown of RNaseH2A inhibited the proliferation of hepatocellular carcinoma cells and induced apoptosis. Knockdown of RNaseH2A induced the high expression of Fas, a key gene for apoptosis, and liver cancer usually features low expression of Fas. After hepatocellular carcinoma cells that were knocked down of RNaseH2A continued were subject to Fas knockdown, hepatocellular carcinoma cell proliferation and apoptosis returned to normal levels. Conclusion: The high expression of RNaseH2A regulates the low expression of Fas, a key gene for apoptosis, thereby inhibiting apoptosis, promoting cell proliferation, and participating in the development of liver cancer.

Balogh J, Victor D, Asham EH, et al, 2016, Hepatocellular Carcinoma: A Review. J Hepatocell Carcinoma, 3(1): 41-53.

Attwa MH, El-Etreby SA, 2015, Guide for Diagnosis and Treatment of Hepatocellular Carcinoma. World J Hepatol, 7(12): 1632-1651.

Kumar M, Panda D, 2014, Role of Supportive Care for Terminal Stage Hepatocellular Carcinoma. J Clin Exp Hepatol, 4(Suppl 3): S130-S139.

Moelling K, Broecker F, 2015, The Reverse Transcriptase-RNase H: From Viruses to Antiviral Defense. Ann NY Acad Sci, 1341(4): 126-135.

Natiq A, Elalaoui SC, Miesch S, et al., 2014, A New Case of De Novo 19p13.2p13.12 Deletion in a Girl with Overgrowth and Severe Developmental Delay. Mol Cytogenet, 7(1): 40.

Flanagan JM, Funes JM, Henderson S, et al., 2009, Genomics Screen in Transformed Stem Cells Reveals RNASEH2A, PPAP2C, and ADARB1 as Putative Anticancer Drug Targets. Mol Cancer Ther, 8(1): 249-260.

Feng S, Cao Z, 2016, Is the Role of Human RNase H2 Restricted to its Enzyme Activity? Prog Biophys Mol Biol, 121(1): 66-73.

Reijns MA, Bubeck D, Gibson LC, et al., 2011, The Structure of the Human RNase H2 Complex Defines Key Interaction Interfaces Relevant to Enzyme Function and Human Disease. J Biol Chem, 286(12): 10530-10539.

Hausen P, Stein H, 1970, Ribonuclease H: An Enzyme Degrading the RNA Moiety of DNA-RNA Hybrids. Eur J Biochem, 14(2): 278-283.

Stein H, Hausen P, 1969, Enzyme from Calf Thymus Degrading the RNA Moiety of DNA-RNA Hybrids: Effect on DNA-Dependent RNA Polymerase. Science, 166(3903): 393-395.

Coté ML, Roth MJ, 2008, Murine Leukemia Virus Reverse Transcriptase: Structural Comparison with HIV-1 Reverse Transcriptase. Virus Res, 134(1-2): 186-202.

Mizuno M, Yasukawa K, Inouye K, 2010, Insight into the Mechanism of the Stabilization of Moloney Murine Leukaemia Virus Reverse Transcriptase by Eliminating RNase H Activity. Biosci Biotechnol Biochem, 74(2): 440-442.

Schultz SJ, Champoux JJ, 2008, RNase H Activity: Structure, Specificity, and Function in Reverse Transcription. Virus Res, 134(1-2): 86-103.

Rice GI, Forte GM, Szynkiewicz M, et al., 2013, Assessment of Interferon-Related Biomarkers in Aicardi-Goutières Syndrome Associated with Mutations in TREX1, RNASEH2A, RNASEH2B, RNASEH2C, SAMHD1, and ADAR: A Case-Control Study. Lancet Neurol, 12(2): 1159-1169.

Orcesi S, La Piana R, Fazzi E, 2009, Aicardi-Goutieres Syndrome. Br Med Bull, 89(1): 183-201.

Crow YJ, 2013, Aicardi-Goutières Syndrome. Handb Clin Neurol, 113(1): 1629-1635.

Crow YJ, Manel N, 2015, Aicardi-Goutières Syndrome and the Type I Interferonopathies. Nat Rev Immunol, 15(7): 429-440.

Williams KA, Lee M, Hu Y, et al, 2014, A Systems Genetics Approach Identifies CXCL14, ITGAX, and LPCAT2 as Novel Aggressive Prostate Cancer Susceptibility Genes. PLoS Genet, 10(11): e1004809.

Tang Z, Li C, Kang B, et al., 2017, GEPIA: A Web Server for Cancer and Normal Gene Expression Profiling and Interactive Analyses. Nucleic Acids Research, 45(1): 98-102.

Yun K, Fidler AE, Eccles MR, et al., 1993, Insulin-Like Growth Factor II and WT1 Transcript Localization in Human Fetal Kidney and Wilms’ Tumor. Cancer Res, 53(21): 5166-5171.

Suswam E, Li Y, Zhang X, et al., 2008, Tristetraprolin Downregulates Interleukin-8 and Vascular Endothelial Growth Factor in Malignant Glioma Cells. Cancer Res, 68(3): 674-682.

Zhang K, Dion N, Fuchs B, et al., 2002, The Human Homolog of Yeast SEP1 is a Novel Candidate Tumor Suppressor Gene in Osteogenic Sarcoma. Gene, 298(2): 121-127.

Rökman A, Ikonen T, Seppälä EH, et al., 2002, Germline Alterations of the RNASEL Gene, a Candidate HPC1 Gene at 1q25, in Patients and Families with Prostate Cancer. Am J Hum Genet, 70(5): 1299-1304.

Krüger S, Silber AS, Engel C, et al., 2005, German Hereditary Non-Polyposis Colorectal Cancer Consortium: Arg462Gln Sequence Variation in the Prostate-Cancer-Susceptibility Gene RNASEL and Age of Onset of Hereditary Non-Polyposis Colorectal Cancer: A Case-Control Study. Lancet Oncol, 6(8): 566-572.

Bartsch DK, Fendrich V, Slater EP, et al., 2005, RNASEL Germline Variants are Associated with Pancreatic Cancer. Int J Cancer, 117(5): 718-722.

Sidrauski C, Walter P, 1997, The Transmembrane Kinase Ire1p is a Site-Specific Endonuclease that Initiates mRNA Splicing in the Unfolded Protein Response. Cell, 90(6): 1031-1039.

Davies MP, Barraclough DL, Stewart C, et al., 2008, Expression and Splicing of the Unfolded Protein Response Gene XBP-1 are Significantly Associated with Clinical Outcome of Endocrine-Treated Breast Cancer. Int J Cancer, 123(1): 85-88.

Sugito N, Ishiguro H, Kuwabara Y, et al., 2006, RNAseN Regulates Cell Proliferation and Affects Survival in Esophageal Cancer Patients. Clin Cancer Res, 12(24): 7322-7328.

Muralidhar B, Goldstein LD, Ng G, et al., 2007, Global Microrna Profiles in Cervical Squamous Cell Carcinoma Depend on Drosha Expression Levels. J Pathol, 212(4): 368-377.

Kaul D, Sikand K, 2004, Defective RNA-Mediated c-myc Gene Silencing Pathway in Burkitt’s Lymphoma. Biochem Biophys Res Commun, 313(3): 552-554.