Identification of renal cell carcinoma specific genes and in vitro confirmation of their expression in ACHN cell line in the presence of Staphylococcus aureus toxin SHE

Authors
Maryam Safarpour-Dehkordi 1
Abbas Doosti 2
Mohammad-Saeid Jami 3
1 PhD Student at Department of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
2 Associate Professor at Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
3 Assistant Professor at Department of Biology, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
10.22034/pjms.2022.700535
Abstract
Introduction: Renal cell carcinoma (RCC) is the most common type of kidney cancer in adults. The aim of the present
study was to obtain specific genes in RCC, based on bioinformatics studies and to evaluate the expression change of candidate genes in ACHN cells after transfection of the seh gene of Staphylococcus aureus.
Material and Methods: The expression correlations of genes involved in RCC were investigated in the TCGA database, and
networks of genes was constructed. In vivo studies were performed in the ACHN cell model, in which the staphylococcal seh gene was cloned into the pcDNA3.1 (+) vector and transfected into ACHN cells. After confirming the correct function of the recombinant vector in these cells and inducing apoptosis, the expression of candidate genes was evaluated.
Results: The results showed the successful identification of the expression correlations of genes involved in apoptosis and lncRNA-mRNA interaction network in RCC. Induction of apoptosis under the influence of toxin expression was confirmed by flow cytometry. Expression of SOX9, SBF2/AS1, MAF and c-Maf genes in cells ordered by recombinant vector was significantly reduced compared to control cells. Also, increased expression of LINC00671 and LINC02499 genes was observed in cells transfected with pcDNA3.1 (+)- seh vector compared to the control group (p<0.05) and This change in expression confirmed the accurate identification of genes involved in RCC cell apoptosis.
Conclusion: The gene networks identified in this study can provide a new perspective for research aimed at identifying new molecular pathways and treating RCC.
Keywords
Kidney Cell Carcinoma
Bioinformatics Analysis
SEH Toxin
ACHN Cell Line
1. Parsa N. Cellular and molecular basis of cancer in humans. Cell Tissue J 2012;2(4):365-76. 2. Athena C, Randolph M. Evolutionary foundations for cancer biology. Evol Appl 2013; 6(1): 144-159. 3. Nabi S, Kessler ER, Bernard B, Flaig TW, Lam ET. Renal cell carcinoma: a review of biology and pathophysiology. F1000Research 2018;7: 1-10. 4. Medina-Rico M, Ramos HL, Lobo M, Romo J, Prada JG. Epidemiology of renal cancer in developing countries: Review of the literature. CUAJ 2018;12(3):E154. 5. Sánchez-Gastaldo A, Kempf E, del Alba AG, Duran I. Systemic treatment of renal cell cancer: a comprehensive review. Cancer Treat Rev 2017;60:77-89. 6. Forbes NS. Engineering the perfect (bacterial) cancer therapy. Nat Rev Cancer 2010;10(11):785-94. 7. Pahle J, Menzel L, Niesler N, Kobelt D, Aumann J, Rivera M, et al. Rapid eradication of colon carcinoma by Clostridium perfringens Enterotoxin suicidal gene therapy. BMC Cancer 2017;17(1):1-4. 8. Liu Y, Chen W, Ali T, Alkasir R, Yin J, Liu G, et al. Staphylococcal enterotoxin H induced apoptosis of bovine mammary epithelial cells in vitro. Toxins 2014;6(12):3552-67. 9. Zhang X, Hu X, Rao X. Apoptosis induced by Staphylococcus aureus toxins. Microbiol 2017;205:19-24. 10. Rudenko NV, Karatovskaya AP, Noskov AN, Shepelyakovskaya AO, Shchannikova MP, Loskutova IV, et al. Immunochemical assay with monoclonal antibodies for detection of staphylococcal enterotoxin H. J Food Drug Anal 2018;26(2):741-50. 11. Petersson K, Pettersson H, Skartved NJ, Walse B, Forsberg G. Staphylococcal enterotoxin H induces Vα-specific expansion of T cells. J Immunol 2003;170(8):4148-54. 12. Hekmatimoghaddam S, Ahmadi T, Pourrajab F, Dehghani Firoozabadi A, Rahmani K. Altered expression of DNMT3A gene in B-cell acute lymphoblastic leukemia by mercaptopurine. JUMS 2021;19(1):0-. 13. Safarpour M, Kazemi Z, Doosti E, Doosti A. Cloning tagD gene from helicobacter pylori in PFLAG-CMV-3 eukaryotic vector to generate a DNA vaccine. JUMS 2016;14(4):43-50. 14. Seim I, Jeffery PL, Thomas PB, Walpole CM, Maugham M, Fung JN, Yap PY, O’Keeffe AJ, Lai J, Whiteside EJ, Herington AC. Multi-species sequence comparison reveals conservation of ghrelin gene-derived splice variants encoding a truncated ghrelin peptide. Endocrine 2016;52(3):609-17. 15. Sotoodeh Jahromi A, Moradzadeh M, Kargar M, Kafilzadeh F, Jamalidoust M. Effect of Crocin on miRNA-15a expression in EBV infected transformed B cell. JUMS 2020;18(3):21-31. 16. Pahle J, Walther W. Bacterial toxins for oncoleaking suicidal cancer gene therapy. Curr S Cancer Gene Ther 2016; 209: 95-110. 17. Chuan T, Li T, Yi C. Identification of CXCR4 and CXCL10 as potential predictive biomarkers in triple negative breast cancer (TNBC). Med Sci Moni 2020;26:e918281-1. 18. Xiao B, Hang J, Lei T, He Y, Kuang Z, Wang L, Chen L, He J, Zhang W, Liao Y, Sun Z. Identification of key genes relevant to the prognosis of ER-positive and ER-negative breast cancer based on a prognostic prediction system. Mol Biol Rep 2019;46(2):2111-9. 19. Safarpour-Dehkordi M, Doosti A, Jami MS. Impacts of the Staphylococcal Enterotoxin H on the Apoptosis and lncRNAs in PC3 and ACHN. Mol Gen Microbiol Virol 2020;35(3):180-8. 20. Liu XH, Sun M, Nie FQ, Ge YB, Zhang EB, Yin DD, et al. LncRNA HOTAIR functions as a competing endogenous RNA to regulate HER2 expression by sponging miR-331-3p in gastric cancer. Mol Cancer 2014;13(1):92.