A novel lncRNA ARST represses glioma progression by inhibiting ALDOA-mediated actin cytoskeleton integrity
Tóm tắt
Glioma is one of the most aggressive malignant brain tumors that is characterized with inevitably infiltrative growth and poor prognosis. ARST is a novel lncRNA whose expression level is significantly decreased in the patients with glioblastoma multiforme. However, the exact mechanisms of ARST in gliomagenesis are largely unknown. The expressions of ARST in the glioma samples and cell lines were analyzed by qRT-PCR. FISH was utilized to detect the distribution of ARST in the glioma cells. CCK-8, EdU and flow cytometry were used to examine cellular viability, proliferation and apoptosis. Transwell and wound-healing assays were performed to determine the migratory and invasive abilities of the cells. Intracranial tumorigenesis models were established to explore the roles of ARST in vivo. RNA pulldown assay was used to examine proteins that bound to ARST. The activities of key enzymes in the glycolysis and production of lactate acid were measured by colorimetry. In addition, RIP, Co-IP, western blot and immunofluorescence were used to investigate the interaction and regulation between ARST, F-actin, ALDOA and cofilin. In this study, we reported that ARST was downregulated in the gliomas. Overexpression of ARST in the glioma cells significantly suppressed various cellular vital abilities such as cell growth, proliferation, migration and invasion. The tumorigenic capacity of these cells in vivo was reduced as well. We further demonstrated that the tumor suppressive effects of ARST could be mediated by a direct binding to a glycolytic enzyme aldolase A (ALDOA), which together with cofilin, keeping the polymerization and depolymerization of actin filaments in an orderly dynamic equilibrium. Upregulation of ARST interrupted the interaction between ALDOA and actin cytoskeleton, which led to a rapid cofilin-dependent loss of F-actin stress fibers. Taken together, it is concluded that ARST performs its function via a non-metabolic pathway associated with ALDOA, which otherwise modifies the morphology and invasive properties of the glioma cells. This has added new perspective to its role in tumorigenesis, thus providing potential target for glioma diagnosis, therapy, and prognosis.
Tài liệu tham khảo
Chen H, Huang Q, Dong J, Zhai DZ, Wang AD, Lan Q. Overexpression of CDC2/CyclinB1 in gliomas, and CDC2 depletion inhibits proliferation of human glioma cells in vitro and in vivo. BMC Cancer. 2008;8(1). https://doi.org/10.1186/1471-2407-8-29.
Yi DA, Hua TX, Lin HY. EGFR gene overexpression retained in an invasive Xenograft model by solid Orthotopic transplantation of human Glioblastoma Multiforme into nude mice. Cancer Investig. 2011;29(3):229–39. https://doi.org/10.3109/07357907.2010.550665.
Salyakina D, Tsinoremas NF. Non-coding RNAs profiling in head and neck cancers. Npj Genomic Medicine. 2016;1(1):15004. https://doi.org/10.1038/Npjgenmed.2015.4.
Rani N, Nowakowski TJ, Zhou HJ, Godshalk SE, Lisi V, Kriegstein AR, et al. A primate lncRNA mediates notch signaling during neuronal development by sequestering miRNA. Neuron. 2016;90(6):1174–88. https://doi.org/10.1016/j.neuron.2016.05.005.
Xu YJ, Wu W, Han Q, Wang YL, Li CC, Zhang PP, Xu HX. New Insights into the Interplay between Non-Coding RNAs and RNA-Binding Protein HnRNPK in Regulating Cellular Functions. Cells-Basel. 2019; Artn 62 https://doi.org/10.3390/Cells8010062
Han MZ, Wang S, Fritah S, Wang X, Zhou WJ, Yang N, et al. Interfering with long non-coding RNA MIR22HG processing inhibits glioblastoma progression through suppression of Wnt/beta-catenin signalling. Brain. 2020;143(2):512–30. https://doi.org/10.1093/brain/awz406.
Tan SK, Pastori C, Penas C, Komotar RJ, Ivan ME, Wahlestedt C, et al. A primate lncRNA mediates notch signaling during neuronal development by sequestering miRNA. Mol Cancer. 2016;90(6):1174–88. https://doi.org/10.1016/j.neuron.2016.05.005.
Li X, Jiang FX, Ge Z, Chen B, Yu J, Xin MJ, et al. Fructose-Bisphosphate Aldolase A Regulates hypoxic adaptation in hepatocellular carcinoma and involved with tumor malignancy. Dig Dis Sci. 2019;64(11):3215–27. https://doi.org/10.1007/s10620-019-05642-2.
Na N, Li H, Xu CF, Miao B, Hong LQ, Huang ZY, et al. High expression of Aldolase a predicts poor survival in patients with clear-cell renal cell carcinoma. Ther Clin Risk Manag. 2017;Volume 13:279–85. https://doi.org/10.2147/Tcrm.S123199.
Du S, Guan ZZ, Hao LH, Song Y, Wang L, Gong LL, Liu L, Qi XY, Hou ZY, Shao SJ. Fructose-Bisphosphate Aldolase A Is a Potential Metastasis-Associated Marker of Lung Squamous Cell Carcinoma and Promotes Lung Cell Tumorigenesis and Migration. PLoS One. 2014; ARTN e85804 https://doi.org/10.1371/journal.pone.0085804
Gizak A, Wisniewski J, Heron P, Mamczur P, Sygusch J, Rakus D. Targeting a moonlighting function of aldolase induces apoptosis in cancer cells. Cell Death Dis. 2019; Artn 712 https://doi.org/10.1038/S41419-019-1968-4
Tokuraku K, Kuragano M, Uyeda TQP. Long-Range and Directional Allostery of Actin Filaments Plays Important Roles in Various Cellular Activities. Int J Mol Sci. 2020; Artn 3209 https://doi.org/10.3390/Ijms21093209
Wang Q, Crevenna AH, Kunze I, Mizuno N. Structural basis for the extended CAP-Gly domains of p150(glued) binding to microtubules and the implication for tubulin dynamics. P Natl Acad Sci USA. 2014;111(31):11347–52. https://doi.org/10.1073/pnas.1403135111.
Carlier MF, Ressad F, Pantaloni D. Control of actin dynamics in cell motility - role of ADF/cofilin. J Biol Chem. 1999;274(48):33827–30. https://doi.org/10.1074/jbc.274.48.33827.
Chang YC, Chiou J, Yang YF, Su CY, Lin YF, Yang CN, et al. Therapeutic targeting of Aldolase a interactions inhibits lung Cancer metastasis and prolongs survival. Cancer Res. 2019:canres.4080.2018. https://doi.org/10.1158/0008-5472.CAN-18-4080.
Gao TH, Gu GY, Tian JX, Zhang R, Zheng XR, Wang YA, et al. LncRNA HSP90AA1-IT1 promotes gliomas by targeting miR-8855p-CDK2 pathway. Oncotarget. 2017. https://doi.org/10.18632/oncotarget.20777.
Gu G, Gao T, Zhang L, Chen X, Pang Q, Wang Y, et al. NKAP alters tumor immune microenvironment and promotes glioma growth via Notch1 signaling. J Exp Clin Cancer Res. 2019;38(1):291. https://doi.org/10.1186/s13046-019-1281-1.
Wisniewski JR, Zougman A, Nagaraj N, Mann M. Universal sample preparation method for proteome analysis. Nat Methods. 2009;6(5):359–62. https://doi.org/10.1038/nmeth.1322.
Peng W, Zhang C, Peng JN, Huang YJ, Peng CF, Tan YQ, et al. Lnc-FAM84B-4 acts as an oncogenic lncRNA by interacting with protein hnRNPK to restrain MAPK phosphatases-DUSP1 expression. Cancer Lett. 2020;494:94–106. https://doi.org/10.1016/j.canlet.2020.08.036.
Kusakabe T, Motoki K, Hori K. Mode of interactions of human aldolase isozymes with cytoskeletons. Arch Biochem Biophys. 1997;344(1):184–93. https://doi.org/10.1006/abbi.1997.0204.
Moncman CL, Peng I, Winkelmann DA. Actin filament structure probed with monoclonal antibodies. Cell Motil Cytoskeleton. 1993;25(1):73–86. https://doi.org/10.1002/cm.970250109.
Li J, Liu Q, Liu ZH, Xia Q, Zhang ZH, Zhang R, Gao TH, Gu GY, Wang YA, Wang D, Chen XY, Yang YH, He D, Xin T. KPNA2 promotes metabolic reprogramming in glioblastomas by regulation of c-myc. J Exp Clin Cancer Res. 2018; Artn 194 https://doi.org/10.1186/S13046-018-0861-9
Dashtimoghadam E, Fahimipour F, Tongas N, Tayebi L. Microfluidic fabrication of microcarriers with sequential delivery of VEGF and BMP-2 for bone regeneration. Sci Rep. 2020;10(1):11764. https://doi.org/10.1038/s41598-020-68221-w.
Lin MF, Jungreis I, Kellis M. PhyloCSF: a comparative genomics method to distinguish protein coding and non-coding regions. Bioinformatics. 2011;27(13):i275–82. https://doi.org/10.1093/bioinformatics/btr209.
Mudge JM, Jungreis I, Hunt T, Gonzalez JM, Wright JC, Kay M, et al. Discovery of high-confidence human protein-coding genes and exons by whole-genome PhyloCSF helps elucidate 118 GWAS loci. Genome Res. 2019;29(12):2073–87. https://doi.org/10.1101/gr.246462.118.
Bie L, Zhao G, Wang YP, Zhang B. Kinesin family member 2C (KIF2C/MCAK) is a novel marker for prognosis in human gliomas. Clin Neurol Neurosurg. 2012;114(4):356–60. https://doi.org/10.1016/j.clineuro.2011.11.005.
Yang P, Li J, Peng CF, Tan YQ, Chen RR, Peng W, Gu Q, Zhou JH, Wang L, Tang JW, Feng YF, Sun YM. TCONS_00012883 promotes proliferation and metastasis via DDX3/YY1/MMP1/PI3K-AKT axis in colorectal cancer. Clin Transl Med. 2020; ARTN e211 https://doi.org/10.1002/ctm2.211
Tanaka K, Takeda S, Mitsuoka K, Oda T, Kimura-Sakiyama C, Maeda Y, Narita A. Structural basis for cofilin binding and actin filament disassembly. Nat Commun. 2018; Artn 1860 https://doi.org/10.1038/S41467-018-04290-W
Hu GQ, Taylor DW, Liu J, Taylor KA. Identification of interfaces involved in weak interactions with application to F-actin-aldolase rafts. J Struct Biol. 2018;201(3):199–209. https://doi.org/10.1016/j.jsb.2017.11.005.
Mizuno K. Signaling mechanisms and functional roles of cofilin phosphorylation and dephosphorylation. Cell Signal. 2013;25(2):457–69. https://doi.org/10.1016/j.cellsig.2012.11.001.
Chung T, Na J, Kim YI, Chang DY, Kim YI, Kim H, et al. Dihydropyrimidine dehydrogenase is a prognostic marker for Mesenchymal stem cell-mediated cytosine Deaminase gene and 5-Fluorocytosine Prodrug therapy for the treatment of recurrent Gliomas. Theranostics. 2016;6(10):1477–90. https://doi.org/10.7150/thno.14158.
Islam SMA, Patel R, Bommareddy RR, Khalid KM, Acevedo-Duncan M. The modulation of actin dynamics via atypical protein kinase-C activated Cofilin regulates metastasis of colorectal cancer cells. Cell Adhes Migr. 2019;13(1):106–20. https://doi.org/10.1080/19336918.2018.1546513.
Dettling S, Stamova S, Warta R, Schnolzer M, Rapp C, Rathinasamy A, et al. Identification of CRKII, CFL1, CNTN1, NME2, and TKT as novel and frequent T-cell targets in human IDH-mutant Glioma. Clin Cancer Res. 2018;24(12):2951–62. https://doi.org/10.1158/1078-0432.CCR-17-1839.
Sun WB, Yan H, Qian CF, Wang CH, Zhao MJ, Liu YC, et al. Cofilin-1 and phosphoglycerate kinase 1 as promising indicators for glioma radiosensibility and prognosis. Oncotarget. 2017. https://doi.org/10.18632/oncotarget.19025.
Zhuang H, Li Q, Zhang XR, Ma XD, Wang Z, Liu Y, et al. Downregulation of glycine decarboxylase enhanced cofilin-mediated migration in hepatocellular carcinoma cells. Free Radical Bio Med. 2018;120:1–12. https://doi.org/10.1016/j.freeradbiomed.2018.03.003.
Yan H, Yang K, Xiao H, Zou YJ, Zhang WB, Liu HY. Over-expression of Cofilin-1 and Phosphoglycerate kinase 1 in Astrocytomas involved in pathogenesis of Radioresistance. Cns Neurosci Ther. 2012;18(9):729–36. https://doi.org/10.1111/j.1755-5949.2012.00353.x.