Evaluation of SCAMP5 Gene Expression and Its Target miR in the Tissue of Patients with Glioblastoma Multiforme

Subject Areas : Journal of Animal Biology

Nasrin Karimi ¹ (Department of Biology, Faculty of Basic Sciences, Islamic Azad University, Damghan Branch, Damghan, Iran)
Hamidreza Kheiri Manjilli ² (Department of Pharmaceutical Nanotechnology, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran)
Vajiheh Zarrinpour ³ (Department of Biology, Faculty of Basic Sciences, Islamic Azad University, Damghan Branch, Damghan, Iran)
Mohammad mahdi Forghanifard ⁴ (1. Department of Biology, Faculty of Basic Sciences, Islamic Azad University, Damghan Branch, Damghan, Iran)

Received: 2021-03-24 Accepted : 2021-05-14 Published : 2022-05-22

Keywords: Real-time PCR, biomarker, Glioblastoma multiforme, SCAMP5,

Abstract :

Glioblastoma multiforme is one of the most malignant and common brain tumors, and after current combination therapy, patient survival is relatively poor and the disease is always fatal. This study was aimed at investigating the expression of SCAMP5 as a possible biomarker in GBM and evaluating the target miR of this gene. 50 tissue samples with GBM form before treatment and 50 healthy tissue samples from the GBM tumor margin were collected as a control. RNA extraction and cDNA synthesis were performed for all samples. Subsequently, a specific primer was designed and the expression of SCAMP5 gene was examined using Real-time PCR. ACTB gene was used as the internal control. Statistical analysis was carried out using Graphpad Prism software version 8. Roc curve was used to evaluate the biomarker value of SCAMP5 gene. Decreased expression of SCAMP5 gene was observed in GBM samples and its expression was compared with the pathological characteristics of patients. Expression of SCAMP5 gene with patients’ age with P-value ˂0.05 was statistically significant. However, the expression of this gene with patients’ gender with P-value ˃ 0.05 was not statistically significant. SCAMP5 was expressed in tumor tissue of patients with GBM much lower than healthy tissue around the tumor. The results revealed that SCAMP5 expression decreased in patients older than 50 years. ROC curve showed that the SCAMP5 gene could have a biomarker value. According to this study, it may be possible to use the expression of this gene as a marker as an early diagnostic method.

References:

1. Bijl N., Thys C., De la Marche W., Devriendt K., Peeters H., Van Geet C., Freson K., 2015. Platelet studies in autism spectrum disorder patients and first-degree relatives. Molecular autism, 6(1): 1-10.


2. Castermans D., Volders K., Crepel A., Backx L., De Vos R., Freson K., Meulemans S., Vermeesch JR., Schrander-Stumpel CT., De Rijk P., Del-Favero J.,2010. SCAMP5, NBEA and AMISYN: three candidate genes for autism involved in secretion of large dense-core vesicles. Human molecular genetics, 19(7): 1368-1378.


3. Dashti S., Ashouri S., Kheirollahi M., 2015. Expression of TERRA in Different Grades of Astrocytom J Isfahan Med Sch, 32(317): 2333-42. [In Persian]


4. Dmitrenko V., Bojko O.I., Shostak K.O., Vitak N.Y., Bukreeva T.V., Rozumenko V.D., Malysheva T.A., Shamayev M.I., Zozulya Y.P., Kavsan V.M., 2007. Characterization of genes, down-regulated in human glioma, potential tumor suppressor genes. Biopolymers and Cell, 23(4): 347.


5. Gaudet P., Livstone MS., Lewis SE., Thomas PD., 2011. Phylogenetic-based propagation of functional annotations within the Gene Ontology consortium. Briefings in Bioinformatics, 12(5): 449-462.


6. Guan N., Huo X., Zhang Z., Zhang S., Luo J., Guo W., 2015. Ginsenoside Rh2 inhibits metastasis of glioblastoma multiforme through Akt-regulated MMP13. Tumor Biology, 36(9): 6789-6795.


7. Guo ST., Jiang CC., Wang GP., Li YP., Wang CY., Guo XY., Yang RH., Feng Y., Wang FH., Tseng H., Thorne R.F., 2013. MicroRNA-497 targets insulin-like growth factor 1 receptor and has a tumour suppressive role in human colorectal cancer. Oncogene, 32(15): 1910-1920.


8. Han C., Chen T., Yang M., Li N., Liu H., Cao X., 2009. Human SCAMP5, a novel secretory carrier membrane protein, facilitates calcium-triggered cytokine secretion by interaction with SNARE machinery. The Journal of Immunology, 182(5): 2986-2996.


9. Hanif F., Muzaffar K., Perveen K., Malhi SM., Simjee SU., 2017. Glioblastoma multiforme: a review of its epidemiology and pathogenesis through clinical presentation and treatment. Asian Pacific Journal of Cancer Prevention, 18(1): 3.


10. Hassani L., Asaadi Tehrani G., Mirza Ahmadi S., 2018. Relationship between LncRNA THRIL expression controlling TNF-alpha pathway in glioblastoma cell line under temozolomide treatment. Tehran University Medical Journal TUMS Publications, 76(7): 469-476. [In Persian]


11. Heo MJ., Kim A., Park C., 2019. Data on molecular characterization and gene expression analysis of secretory carrier-associated membrane protein 5 (SCAMP5) from the red sea bream (Pagrus major). Data in Brief, 25: 103901.


12. Holland EC., 2000. Glioblastoma multiforme: the terminator. Proceedings of the National Academy of Sciences, 97(12): 6242-6244.


13. Huang Q., Li H., Dai X., Zhao D., Guan B., Xia W., 2019. miR‑497 inhibits the proliferation and migration of A549 non‑small‑cell lung cancer cells by targeting FGFR1. Molecular Medicine Reports, 20(4): 3959-3967.


14. Jiao X., Morleo M., Nigro V., Torella A., D’Arrigo S., Ciaccio C., Pantaleoni C., Gong P., Grand K., Sanchez-Lara PA., Krier J., 2020. Identification of an identical de novo SCAMP5 missense variant in four unrelated patients with seizures and severe neurodevelopmental delay. Frontiers in Pharmacology, 11: 1-8.


15. Jonas S., Izaurralde E., 2015. Towards a molecular understanding of microRNA-mediated gene silencing. Nature Reviews of Genetics, 16(7): 421-433.


16. Kanehisa M., Goto S., 2000. KEGG: kyoto encyclopedia of genes and genomes. Nucleic Acids Research, 28(1):27-30.


17. Leone P., González M.B., Elosua C., Gómez-Moreta J.A., Lumbreras E., Robledo C., Santos-Briz A., Valero J.M., de la Guardia R.D., Gutiérrez N.C., Hernández J.M., 2012. Integration of global spectral karyotyping, CGH arrays, and expression arrays reveals important genes in the pathogenesis of glioblastoma multiforme. Annals of Surgical Ooncology, 19(7): 2367-2379.


18. LI S., Guo W., Gao Y., Liu Y., 2015, Ginsenoside Rh2 inhibits growth of glioblastoma multiforme through mTor. Tumor Biology, 36(4): 2607-2612.


19. Lin P., Williams W.P., Luu Y., Molday R.S., Orlowski J., Numata M., 2005. Secretory carrier membrane proteins interact and regulate trafficking of the organellar (Na+, K+)/H+ exchanger NHE7. Journal of Cell Science, 1; 118(9):1885-97.


20. Liu C., Su C., Design strategies and application progress of therapeutic exosomes. Theranostics, 9(4): 1015.


21. Luo Y., Bao X., Zheng S., Gu T., Mao S., Liu S., Sun J., Huang M., Zhang L.,2020. A potential risk factor of essential hypertension in case-control study: MicroRNAs miR-10a-5p. Clinical and Experimental Hypertension, 42(1): 36-42.


22. Noh JY., Lee H., Song S., Kim NS., Im W., Kim M., Seo H., Chung CW., Chang JW., Ferrante R., Yoo Y.J., 2009. SCAMP5 links endoplasmic reticulum stress to the accumulation of expanded polyglutamine protein aggregates via endocytosis inhibition. Journal of Biological Chemistry, 284(17): 11318-11325.‏


23. Park D., Lee U., Cho E., Zhao H., Kim JA., Lee BJ., Regan P., Ho WK., Cho K., Chang S., 2018. Impairment of release site clearance within the active zone by reduced SCAMP5 expression causes short-term depression of synaptic release. Cell reports, 22(12): 3339-3350.


24. Qian T., Cheng Z., Quan L., Zeng T., Cui L., Liu Y., Si C., Huang W., Dai Y., Chen J., Liu L., 2020. Prognostic role of SCAMP family in acute myeloid leukemia. The Pharmacogenomics Journal, 20(4):595-600.


25. Regazzo G., Terrenato I., Spagnuolo M., Carosi M., Cognetti G., Cicchillitti L., Sperati F., Villani V., Carapella C., Piaggio G., Pelosi A., 2016. A restricted signature of serum miRNAs distinguishes glioblastoma from lower grade gliomas. Journal of Experimental & Clinical Cancer Research, 35(1): 124.‏


26. Sánchez-Valle J, Tejero H, Ibáñez K, Portero J.L, Krallinger M, Al-Shahrour F, Tabarés-Seisdedos R, Baudot A, Valencia A., 2017. A molecular hypothesis to explain direct and inverse co-morbidities between Alzheimer ’s disease, Glioblastoma and Lung cancer. Scientific Reports, 7(1): 1-12.


27. Shabani M., Mohammad Ganji S., Salahshouri Far I., 2020. Investigation of Long Non-coding RNA HOX A11-AS Expression in Iranian Patients with Glioblastoma: A Quantitative Study. Journal of Qom University of Medical Science, 14(5):22-29. [In Persian]


28. Shahsavani N., Mazaheri M., 2017. The Effect of Resveratrol on Expression of microRNA34a in U87MG Cancerous Cells Line. Journal of Isfahan Medical School, 35(429): 525-30. [In Persian]


29. Shen L., Lin Y., Sun Z., Yuan X., Chen L., Shen B., 2016. Knowledge-guided bioinformatics model for identifying autism spectrum disorder diagnostic MicroRNA biomarkers. Scientific reports, 6(1): 1-9.


30. Sherman BT., Lempicki RA., 2009. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nature Protocols, 4(1): 44-57.


31. Stigbrand T., 2016. Ginsenoside Rh2 inhibits invasiveness of glioblastoma through modulation of VEGF-A. Tumor Biol,  37:15477-15482.


32. Tabuchi S., 2015. The autotaxin-lysophosphatidic acid–lysophosphatidic acid receptor cascade: proposal of a novel potential therapeutic target for treating glioblastoma multiforme. Lipids in health and disease, 14(1): 1-9.


33. The human gene database. Gene cards.2021.availablefrom;https://www.genecards.org/cgi-bin/carddisp.pl?gene=SCAMP5& keywords=SCAMP5.


34. Tian F., Zhan Y., Zhu W., Li J., Tang M., Chen X., Jiang J., 2019. MicroRNA-497 inhibits multiple myeloma growth and increases susceptibility to bortezomib by targeting Bcl-2 Corrigendum in/10.3892/ijmm. 2019.4297. International Journal of Molecular Medicine, 43(2): 1058-1066.


35. Upton A., Arvanitis TN., 2012. Investigating survival prognosis of glioblastoma using evolutional properties of gene networks. In2012 IEEE 12th International Conference on Bioinformatics and Bioengineering (BIBE), pp: 466-471.


36. Xu F., Li F., Zhang W., Jia P., 2015. Growth of glioblastoma is inhibited by miR-133-mediated EGFR suppression. Tumor Biology, 36(12): 9553-9558.‏


37.  Xu N., Wu YP., Yin HB., Xue XY., Gou X., 2018. Molecular network-based identification of competing endogenous RNAs and mRNA signatures that predict survival in prostate cancer. Journal of Translational Medicine, 16(1): 274.


38. Yang Y., Qin M., Bao P., Xu W., Xu J., 2017. Secretory carrier membrane protein 5 is an autophagy inhibitor that promotes the secretion of α-synuclein via exosome. PloS One, 12(7): e0180892.


39. Zhao H., Kim Y., Park J., Park D., Lee SE., Chang I., Chang S., 2014. SCAMP5 plays a critical role in synaptic vesicle endocytosis during high neuronal activity. Journal of Neuroscience, 34(30): 10085-10095.


40. Zheng JC., Tham CT., Keatings K., Fan S., Liou AY., Numata Y., Allan D., Numata M., 2014. Secretory carrier membrane protein (SCAMP) deficiency influences behavior of adult flies. Frontiers in Cell and Developmental Biology, 2: 64.


41. Zhong H., Yang J., Zhang B., Wang X., Pei L., Zhang L., Lin Z., Wang Y., Wang C., 2018. LncRNA GACAT3 predicts poor prognosis and promotes cell proliferation in breast cancer through regulation of miR-497/CCND2. Cancer Biomarkers, 22(4): 787-797.


42. Zhu W., Zhu D., Lu S., Wang T., Wang J., Jiang B., Shu Y., Liu P., 2012. miR-497 modulates multidrug resistance of human cancer cell lines by targeting BCL2. Medical Oncology, 29(1): 384-391.