Background
Methods for identifying lncRNAs
Serial analysis of gene expression
Tiling arrays
Cap analysis of gene expression
RNA-sequencing
General strategies for investigating the cellular functions of lncRNAs
Guilt by association
Loss-of-function study
Genetic manipulation approaches targeting lncRNAs
RNA interference
Antisense oligonucleotides
CRISPR-Cas system
Silencing of an lncRNA gene by partial or complete deletion of its genomic locus
Ablation of lncRNA expression by knock-in strategy
CRISPR interference/activation
CRISPR-Cas13 system—a potential tool for targeting lncRNA
Techniques for investigating molecular interaction between lncRNAs and other biomolecules
Functional involvement of lncRNAs | Techniques or methods used | References |
---|---|---|
LncRNA-protein interaction | • RNA immunoprecipitation (RIP) • High-throughput sequencing of RNA isolated by cross-linking immunoprecipitation (HITS-CLIP) • Photoactivatable ribonucleotide-enhanced cross-linking and immunoprecipitation (PAR-CLIP) | |
LncRNA-DNA interaction | • Chromatin isolation by RNA purification (ChIRP) • RNA antisense purification (RAP) • Capture hybridization analysis of RNA targets (CHART) | |
LncRNA-RNA interaction | • RNA antisense purification followed by RNA sequencing (RAP-RNA) • Cross-linking, ligation and sequencing of hybrids (CLASH) | |
LncRNA localization | • RNA single-molecule fluorescence in situ hybridization (RNA smFISH) |
Techniques for studying lncRNA-protein interaction
RNA immunoprecipitation
High-throughput sequencing of RNA isolated by cross-linking immunoprecipitation
Photoactivatable ribonucleotide-enhanced cross-linking and immunoprecipitation
Techniques for studying lncRNA-DNA interaction
Chromatin isolation by RNA purification
RNA antisense purification
Capture hybridization analysis of RNA targets
Techniques for studying lncRNA-RNA interaction
RNA antisense purification followed by RNA sequencing
Cross-linking, ligation, and sequencing of hybrids
Technique for studying lncRNA localization
RNA single-molecule fluorescence in situ hybridization
Functional targets for probing biological effects of lncRNAs in blood cancer cells
Cell proliferation
Disease type | lncRNA involved | Cell proliferation approach | Mechanism/Effect on cell proliferation | Gene manipulation system | References |
---|---|---|---|---|---|
AML | CASC15 | MTS assay | CASC15 expression may limit cell proliferation by regulation SOX4 expression | siRNA | [93] |
AML | CCAT1 | CKK-8 assay | Promotes cell proliferation by sequestering tumor suppressive miR155 | shRNA | [89] |
AML | MEG3 | MTT assay | Suppresses cell proliferation through inducing G0/G1 cell cycle arrest | siRNA | [91] |
AML | NEAT1 | CCK-8 assay | Modulates cell proliferation by regulating miR-23a-3p/SMC1A | pcDNA3.1-NEAT1 (overexpression of NEAT1) | [90] |
AML | TUG1 | CKK-8 assay | Increases cell proliferation through targeting AURKA | Lentiviral vector-mediated gene manipulation | [92] |
AML | UCA1 | Trypan Blue exclusion assay | Sustains cell proliferation by repressing p27kip1 expression | shRNA | [87] |
ALL | LINC-PINT | MTS assay | Overexpressed linc-PINT decreases cell proliferation through apoptosis activation and cell cycle arrest at G2/M phase | Overexpression of linc-PINT by linc-PINT-pCDNA3 | [94] |
ALL | NALT | CKK-8 assay | Upexpressed NALT promotes cell proliferation through interacting with NOTCH signaling pathway | shRNA | [86] |
APL | PVT1 | CKK-8 assay | Promotes cell proliferation by MYC | siRNA | [95] |
CML | HULC | MTT assay | Promotes cell proliferation by regulating PI3K/AKT signaling pathway | shRNA | [97] |
CML | MEG3 | MTT assay | Overexpressed MEG3 inhibits cell proliferation by sponging miR-21 | pLVX-hMEG3-ZsGreen-Puro lentiviral overexpression vector | [96] |
CML | MEG3 | CCK-8 assay | Overexpressed MEG3 inhibits cell proliferation by inhibiting miR-184 | siRNA | [188] |
CML | PLIN2 | MTT assay | Overexpressed PLIN2 promotes cell proliferation through activating GSK-3β and β-catenin | shRNA | [98] |
CLL | BM742401 | MTT assay | Overexpressed BM742401 inhibits cell proliferation through caspase-9 dependent intrinsic pathway | ASO | [88] |
Cell cycle regulation
Name of lncRNA | Effects on cell cycle stage(s) | References |
---|---|---|
LncRNA-HEIH | Suppresses p16. p21, p27, and p57 transcription with PRC2 (G0/G1) | [109] |
MEG3 | Suppresses cyclin D1 and induction of cell cycle arrest in G0/G1 phase | [110] |
ANRIL | Suppresses CDK inhibitors encoded by the INK4 locus Suppresses transcription of p14, p15, and p16 in DNA damage response (G1) | |
HOTAIR | Regulates expression of cell cycle regulators such as cyclin D1, cyclin E, CDK2, CDK4, E2F1 (G1/S) | |
HOXA11-AS | Suppresses CDK inhibitors p16, p21, p27, and Rb protein (G1/S) | [40] |
NcRNACCND1 | Suppresses transcription of cyclin D1 (G1/S) | [111] |
PANDA | Suppresses CDK inhibitor p21 produced from CDKN1A locus Suppresses transcription of p18 (G1/S) | |
MALAT1 | Regulates cell cycle via interaction with hnRNP C Promotes cell-cycle regulators such as cyclin A2 and B1 (G1 and G2/M) |
Hematopoiesis
lncRNA | Observations in lineage differentiation | References |
---|---|---|
HOTAIRM1 | Expression shows myeloid lineage specificity and increases during granulocytic differentiation | |
LINC00173 | Specifically expressed in mature granulocytes; controls differentiation of myeloid progenitor cells towards granulocytes | [24] |
NEAT1 | Highly expressed in APL cells; NEAT1 depletion stopped ATRA-induced granulocytic differentiation | [116] |
LncRNAp53int1 | Expressed in undifferentiated human myeloid leukemia cells and greatly reduced during differentiation towards monocytes and macrophages | [117] |
Lnc-MC | Increased expression promotes differentiation from monocytes to macrophage through sequestering miR-199a-5p | [118] |
Lnc-DC | Exclusively expressed in dendritic cells (DCs); knockdown study showed its involvement in DC differentiation | [119] |
EGO | Highly expressed in mature eosinophils; knockdown of EGO influenced the expression of regulators in eosinophilopoiesis | [120] |
PU.1 AS | Negatively regulates the mRNA translation of the master hematopoietic transcription factor PU.1 | [121] |
Resistance to anti-cancer drug
lncRNA | Sample source | Functional involvement and mechanism of action lncRNAs in drug resistance | Approaches for lncRNAs characterization in drug resistance | References | ||
---|---|---|---|---|---|---|
Cell proliferation/cytotoxicity, cell viability assay | Manipulation approaches for lncRNAs | Mechanism characterization approach | ||||
UCA1 | Imatinib-resistant cell lines | Modulates imatinib resistance by acting as a ceRNA against miR-16 | CCK-8 assay | siRNAs | RIP assay, Dual-luciferase reporter assay | [123] |
SNHG5 | Patient samples, imatinib-resistant cell lines | Promotes imatinib resistance through acting as ceRNA against miR-205-5p | MTT assay | siRNAs | RIP assay, Luciferase reporter assay | [125] |
HOTAIR | Multidrug-resistant patient samples, imatinib-resistant cell lines | Modulates MDR to imatinib resistance through activating PI3K/Akt-dependent pathway | MTT assay, Annexin V/propidium iodide (PI) staining assay | siRNAs | – | [126] |
MEG3 | Patient samples, imatinib-resistant cell lines | Inhibits imatinib resistance by suppressing miR-21 | CCK-8 assay, Annexin V-FITC/PI Apoptosis Detection Kit | Overexpression | Luciferase reporter assay | [127] |
Potential involvement in leukemia-induced angiogenesis
Cancer type | LncRNA | Functional involvement of lncRNAs in angiogenesis | References | |
---|---|---|---|---|
Disease phenotype | Mechanism of action | |||
Gastric cancer | OR3A4 | Promotes cell proliferation, migration, invasion, tumorigenesis, angiogenesis | Increases VEGF-C and MMP9 expression | [143] |
MALAT1 | Promotes tumorigenicity and metastasis | Regulation of VE-cadherin/β-catenin complex and ERK/MMP and FAK/paxillin signaling pathways | [144] | |
PVT1 | Induce angiogenesis within tumors | Mediates angiogenesis via evoking the STAT3/VEGFA signaling axis | [145] | |
CASC2 | Inhibits cell invasion and angiogenesis | – | [146] | |
Glioblastoma | TUG1 | Promotes cell proliferation, migration and angiogenesis Promotes blood-tumor barrier permeability and angiogenesis | Increases VEGFA expression through downregulation of miRNA-299 | [63] |
XIST | Inhibition of FOXC1 and ZO-2 by upregulating miR-137 | [147] | ||
Glioma | HULC | Promotes cell proliferation and angiogenesis | Upregulation of ESM-1 through PI3K/Akt/mTOR signaling pathway | [148] |
H19 | Promotes angiogenesis | Increases the VASH2 expression through overexpressing miRNA-29a | [149] | |
Hepatocellular carcinoma | HULC | Promotes cell proliferation and angiogenesis | Upregulation of SPHK1through miRNA-107/E2F1/SPHK1 signaling pathway | [150] |
MVIH | Promotes cell growth and metastasis | Inhibition of PGK1 secretion | [151] | |
Hepatoblastoma | TUG1 | Promotes cell proliferation, migration and angiogenesis | Increases VEGFA expression by miR-34a-5p downregulation | [152] |
CRNDE | Promotes tumor growth and tumor angiogenesis | Regulates mTOR signaling pathways | [153] | |
Nasopharyngeal carcinoma | HOTAIR | Promotes cell growth and angiogenesis | Upregulation of VEGFA and Ang2 by GRP78 | [56] |
Neuroblastoma | MALAT1 | Promotes angiogenesis | Upregulation of FGF2 expression | [154] |
Diabetes | MIAT | Promotes ocular angiogenesis | Upregulation of VEGF by miRNA-150-5p | [155] |
Pituitary adenomas | MEG3 | Suppresses tumor cell proliferation and angiogenesis | Suppression of VEGF signaling pathway | [156] |