Abstract
The purpose of this study was to investigate the impact of SAM- and SH3-domain containing 1 (SASH1) on the biological behavior of glioma cells, including its effects on cellular growth, proliferation, apoptosis, invasion, and metastasis, and thereby to provide an experimental basis for future therapeutic treatments. A pcDNA3.1-SASH1 eukaryotic expression vector was constructed and transfected into the U251 human glioma cell line. Using the tetrazolium-based colorimetric (MTT) assay, flow cytometry analyses, transwell invasion chamber experiments, and other methods, we examined the impact of SASH1 on the biological behaviors of U251 cells, including effects on viability, cell cycle, apoptosis, and invasion. Furthermore, the effect of SASH1 on the expression of cyclin D1, caspase-3, matrix metalloproteinase (MMP)-2, MMP-9, and other proteins was observed. Compared to the empty vector and blank control groups, the pcDNA3.1-SASH1 group of U251 cells exhibited significantly reduced cell viability, proliferation, and invasion (p < 0.05), although there was no difference between the empty vector and blank control groups. The pcDNA3.1-SASH1 group demonstrated a significantly higher apoptotic index than did the empty vector and blank control groups (p < 0.05), and the percentage of apoptotic cells was similar between the empty vector and blank control groups. In addition, the pcDNA3.1-SASH1 group expressed significantly lower protein levels of cyclin D1 and MMP-2/9 compared to the control and empty vector groups (p < 0.05) and significantly higher protein levels of caspase-3 than the other two groups (p < 0.05). Cyclin D1, caspase-3, and MMP-2/9 expression was unchanged between the empty vector and blank control groups. SASH1 gene expression might be related to the inhibition of the growth, proliferation, and invasion of U251 cells and the promotion of U251 cells apoptosis.
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References
Chamberlain MC. Temozolomide: therapeutic limitations in the treatment of adult high-grade gliomas. Exp Rev Neurother. 2010;10(10):1537–44.
Ermoian R, Ladra M, Patel S. Children’s Oncology Group L991 final study report: establishing an important benchmark for assessing late effects of trimodality care of pediatric patients treated for high grade gliomas. Transl Pediatr. 2012;1(1):3–5.
Iwami K, Natsume A, Wakabayashi T. Cytokine networks in glioma. Neurosurg Rev. 2011;34(3):253–63. discussion 263-254.
Ferguson SD. Malignant gliomas: diagnosis and treatment. Dis Mon. 2011;57(10):558–69.
Rainov NG, Heidecke V. Clinical development of experimental therapies for malignant glioma. Sultan Qaboos Univ Med J. 2011;11(1):5–28.
Tykocki T, Michalik R, Bonicki W, Nauman P. Fluorescence-guided resection of primary and recurrent malignant gliomas with 5-aminolevulinic acid. Preliminary results. Neurol Neurochir Pol. 2012;46(1):47–51.
Prasanna PGS, Stone HB, Wong RS, Capala J, Bernhard EJ, Vikram B, Coleman CN. Normal tissue protection for improving radiotherapy: where are the gaps? Transl Cancer Res. 2012;1(1):35–48.
Sherman JH, Hoes K, Marcus J, Komotar RJ, Brennan CW, Gutin PH. Neurosurgery for brain tumors: update on recent technical advances. Curr Neurol Neurosci Rep. 2011;11(3):313–9.
Park SH, Zhu Y, Ozden O, Kim HS, Jiang H, Deng CX, Gius D, Vassilopoulos A. SIRT2 is a tumor suppressor that connects aging, acetylome, cell cycle signaling, and carcinogenesis. Transl Cancer Res. 2012;1(1):15–21.
Candolfi M, Kroeger KM, Muhammad AK, Yagiz K, Farrokhi C, Pechnick RN, Lowenstein PR, Castro MG. Gene therapy for brain cancer: combination therapies provide enhanced efficacy and safety. Curr Gene Ther. 2009;9(5):409–21.
Ritter H, Antonova L, Mueller CR. The unliganded glucocorticoid receptor positively regulates the tumour suppressor gene BRCA1 through GABP beta. Mol Cancer Res. 2012;10(4):558–69.
Muley T, Herth FJF, Schnabel P, Dienemann H, Meister M. From tissue to molecular phenotyping: pre-analytical requirements Heidelberg experience. Transl Lung Cancer Res. 2012;1(2):111–21.
Alexiou GA, Voulgaris S. The role of the PTEN gene in malignant gliomas. Neurol Neurochir Pol. 2010;44(1):80–6.
Kim YJ, Cho YE, Kim YW, Kim JY, Lee S, Park JH. Suppression of putative tumour suppressor gene GLTSCR2 expression in human glioblastomas. J Pathol. 2008;216(2):218–24.
Qu M, Jiao H, Zhao J, Ren ZP, Smits A, Kere J, Nister M. Molecular genetic and epigenetic analysis of NCX2/SLC8A2 at 19q13.3 in human gliomas. Neuropathol Appl Neurobiol. 2010;36(3):198–210.
Rajan N, Elliott R, Clewes O, Mackay A, Reis-Filho JS, Burn J, Langtry J, Sieber-Blum M, Lord CJ, Ashworth A. Dysregulated TRK signalling is a therapeutic target in CYLD defective tumours. Oncogene. 2011;30(41):4243–60.
Naguib A, Cooke JC, Happerfield L, Kerr L, Gay LJ, Luben RN, Ball RY, Mitrou PN, McTaggart A, Arends MJ. Alterations in PTEN and PIK3CA in colorectal cancers in the EPIC Norfolk study: associations with clinicopathological and dietary factors. BMC Cancer. 2011;11:123.
Pustisek N, Situm M. UV-radiation, apoptosis and skin. Coll Antropol. 2011;35 Suppl 2:339–41.
Marcel V, Dichtel-Danjoy ML, Sagne C, Hafsi H, Ma D, Ortiz-Cuaran S, Olivier M, Hall J, Mollereau B, Hainaut P, Bourdon JC. Biological functions of p53 isoforms through evolution: lessons from animal and cellular models. Cell Death Differ. 2011;18(12):1815–24.
Zeller C, Hinzmann B, Seitz S, Prokoph H, Burkhard-Goettges E, Fischer J, Jandrig B, Schwarz LE, Rosenthal A, Scherneck S. SASH1: a candidate tumor suppressor gene on chromosome 6q24.3 is downregulated in breast cancer. Oncogene. 2003;22(19):2972–83.
Aviv T, Lin Z, Lau S, Rendl LM, Sicheri F, Smibert CA. The RNA-binding SAM domain of Smaug defines a new family of post-transcriptional regulators. Nat Struct Biol. 2003;10(8):614–21.
Koch CA, Anderson D, Moran MF, Ellis C, Pawson T. SH2 and SH3 domains: elements that control interactions of cytoplasmic signaling proteins. Science. 1991;252(5006):668–74.
Claudio JO, Zhu YX, Benn SJ, Shukla AH, McGlade CJ, Falcioni N, Stewart AK. HACS1 encodes a novel SH3-SAM adaptor protein differentially expressed in normal and malignant hematopoietic cells. Oncogene. 2001;20(38):5373–7.
Kim CA, Gingery M, Pilpa RM, Bowie JU. The SAM domain of polyhomeotic forms a helical polymer. Nat Struct Biol. 2002;9(6):453–7.
Kuribayashi K, Nakamura K, Tanaka M, Sato T, Kato J, Sasaki K, Takimoto R, Kogawa K, Terui T, Takayama T, Onuma T, Matsunaga T, Niitsu Y. Essential role of protein kinase C zeta in transducing a motility signal induced by superoxide and a chemotactic peptide, fMLP. J Cell Biol. 2007;176(7):1049–60.
Martini M, Gnann A, Scheikl D, Holzmann B, Janssen KP. The candidate tumor suppressor SASH1 interacts with the actin cytoskeleton and stimulates cell-matrix adhesion. Int J Biochem Cell Biol. 2011;43(11):1630–40.
Alcock HE, Stephenson TJ, Royds JA, Hammond DW. Analysis of colorectal tumor progression by microdissection and comparative genomic hybridization. Genes Chromosomes Cancer. 2003;37(4):369–80.
Lemeta S, Salmenkivi K, Pylkkanen L, Sainio M, Saarikoski ST, Arola J, Heikkila P, Haglund C, Husgafvel-Pursiainen K, Bohling T. Frequent loss of heterozygosity at 6q in pheochromocytoma. Hum Pathol. 2006;37(6):749–54.
Barghorn A, Speel EJ, Farspour B, Saremaslani P, Schmid S, Perren A, Roth J, Heitz PU, Komminoth P. Putative tumor suppressor loci at 6q22 and 6q23-q24 are involved in the malignant progression of sporadic endocrine pancreatic tumors. Am J Pathol. 2001;158(6):1903–11.
Rimkus C, Martini M, Friederichs J, Rosenberg R, Doll D, Siewert JR, Holzmann B, Janssen KP. Prognostic significance of downregulated expression of the candidate tumour suppressor gene SASH1 in colon cancer. Br J Cancer. 2006;95(10):1419–23.
Das SK, Bhutia SK, Kegelman TP, Peachy L, Oyesanya RA, Dasgupta S, Sokhi UK, Azab B, Dash R, Quinn BA, Kim K, Barral PM, Su ZZ, Boukerche H, Sarkar D, Fisher PB. MDA-9/syntenin: a positive gatekeeper of melanoma metastasis. Front Biosci. 2012;17:1–15.
Chetty C, Vanamala SK, Gondi CS, Dinh DH, Gujrati M, Rao JS. MMP-9 induces CD44 cleavage and CD44 mediated cell invasion in glioblastoma xenograft cells. Cell Signal. 2012;24(2):549–59.
Sen T, Chatterjee A. Epigallocatechin-3-gallate (EGCG) downregulates EGF-induced MMP-9 in breast cancer cells: involvement of integrin receptor alpha5beta1 in the process. Eur J Nutr. 2011;50(6):465–78.
Shen KH, Hung SH, Yin LT, Huang CS, Chao CH, Liu CL, Shih YW. Acacetin, a flavonoid, inhibits the invasion and invasion of human prostate cancer DU145 cells via inactivation of the p38 MAPK signaling pathway. Mol Cell Biochem. 2010;333(1–2):279–91.
Chen YY, Chiang SY, Lin JG, Ma YS, Liao CL, Weng SW, Lai TY, Chung JG. Emodin, aloe-emodin and rhein inhibit invasion and invasion in human tongue cancer SCC-4 cells through the inhibition of gene expression of matrix metalloproteinase-9. Int J Oncol. 2010;36(5):1113–20.
Kim A, Kim MJ, Yang Y, Kim JW, Yeom YI, Lim JS. Suppression of NF-kappaB activity by NDRG2 expression attenuates the invasive potential of highly malignant tumor cells. Carcinogenesis. 2009;30(6):927–36.
Bausero MA, Bharti A, Page DT, Perez KD, Eng JW, Ordonez SL, Asea EE, Jantschitsch C, Kindas-Muegge I, Ciocca D, Asea A. Silencing the hsp25 gene eliminates invasion capability of the highly metastatic murine 4T1 breast adenocarcinoma cell. Tumour Biol. 2006;27(1):17–26.
Kanno T, Kamba T, Yamasaki T, Shibasaki N, Saito R, Terada N, Toda Y, Mikami Y, Inoue T, Kanematsu A, Nishiyama H, Ogawa O, Nakamura E. JunB promotes cell invasion and angiogenesis in VHL-defective renal cell carcinoma. Oncogene. 2011;31(25):3098–110.
Yang X, Zhang P, Ma Q, Kong L, Li Y, Liu B, Lei D. EMMPRIN contributes to the in vitro invasion of human salivary adenoid cystic carcinoma cells. Oncol Rep. 2012;27(4):1123–7.
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Yang, L., Liu, M., Gu, Z. et al. Overexpression of SASH1 related to the decreased invasion ability of human glioma U251 cells. Tumor Biol. 33, 2255–2263 (2012). https://doi.org/10.1007/s13277-012-0487-z
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DOI: https://doi.org/10.1007/s13277-012-0487-z