Journal: OncoTargets and therapy
Article Title: lncRNA TMPO-AS1 Exerts Oncogenic Roles in HCC Through Regulating miR-320a/SERBP1 Axis
doi: 10.2147/OTT.S250355
Figure Lengend Snippet: TMPO-AS1 serves as a sponge of miR-320a. ( A ) The localization of TMPO-AS1 in cells (SNU-387 and HCCLM3) was confirmed by nuclear-cytoplasmic fractionation. ( B ) The potential binding sites between miR-320a and TMPO-AS1 were hypothesized using bioinformatics analysis starBase v.2.0 database. ( C ) The targeted binding effects between miR-320a and TMPO-AS1-wt or TMPO-AS1-mut in SNU-387 and HCCLM3 cells were detected using luciferase reporter assay. ** P <0.01 compared with miR-NC. ( D ) RIP assay was conducted to confirm the binding ability between TMPO-AS1 and miR-320a. Both TMPO-AS1 and miR-320a were significantly enriched in Ago2 immunoprecipitate. ** P <0.01 compared with anti-IgG. ( E ) RNA-FISH analysis showed that miR-320a co-localizes with TMPO-AS1 in SNU-387 and HCCLM3 cells. ( F ) The relative expression of miR-320a in SNU-387 and HCCLM3 cell transfected with sh-TMPO-AS1 or pc-TMPO-AS1 was measured by RT-qPCR. ** P <0.01 compared with sh-NC. ( G ) The levels of miR-320a in HCC cells and LO2 cells were determined by qRT-PCR assay. ** P <0.01 compared with LO2. ( H ) Cell viability of SNU-387 and HCCLM3 cell after miR-320a upregulation was conducted by performing CCK-8 assay. ( I ) The proliferation of miR-320a overexpressed SNU-387 and HCCLM3 cells was examined by colony formation assay. ( J ) The migration of miR-320a overexpressed SNU-387 and HCCLM3 cells was examined by wound healing assay. ( K ) The invasion of miR-320a overexpressed SNU-387 and HCCLM3 cell was examined by Transwell invasion assay. ** P <0.01 compared with control.
Article Snippet: HCC cells (HepG2, SNU-387, HCCLM3, SMMC-7721, Huh7) and normal human hepatic cell line, LO2 were purchased from Jiangsu KeyGEN BioTECH (Nanjing, Jiangsu, China) and were maintained in RPMI-1640 or DMEM containing 10% FBS at 37°C in a 5% CO 2 incubator.
Techniques: Fractionation, Binding Assay, Luciferase, Reporter Assay, Expressing, Transfection, Quantitative RT-PCR, CCK-8 Assay, Colony Assay, Migration, Wound Healing Assay, Transwell Invasion Assay, Control