Long non-coding RNA AL139002.1 promotes gastric cancer development by sponging microRNA-490-3p to regulate Hepatitis A Virus Cellular Receptor 1 expression

Figures & data

Table

LncRNA AL139002.1	Forward, 5′-TTCTCTGTGTCAGGCCTTTGA-3′
LncRNA AL139002.1	Reverse, 5′-GCAGGAACAGGCCATTTTCA-3′
miR-490-3p	Forward, 5′-CGCAACCTGGAGGACTCC-3′
miR-490-3p	Reverse, 5′-AGTGCAGGGTCCGAGGTATT-3′
HAVCR1	Forward, 5′-TGGTGGGAGATAGAGGAAGCAT-3′
HAVCR1	Reverse, 5′-GATCAGCGTTCAGATCCAGG-3′
GAPDH	Forward, 5′-CTCAGACACCATGGGGAAGGTGA-3′
GAPDH	Reverse, 5′- ATGATCTTGAGGCTGTTGTCATA-3′
U6	Forward, 5′-CTCGCTTCGGCAGCACA-3′
U6	Reverse, 5′-AACGCTTCACGAATTTGCGT-3′

Figure 1. Bioinformatics prediction of lncRNA AL139002.1 and targeting miRNA/mRNA and validation. (a-f) Volcano plot and Heatmap of DElncRNAs, DEmiRNAs, and DEmRNAs in the TCGA database. (g) TCGA and miRcode database consistently predicted five mRNAs interacted with lncRNA. (h) LncRNA-miRNA-mRNA regulatory network. (i, j) Kaplan-Meier survival analysis showed the relationship between lncRNA AL139002.1 or HAVCR1 expression with the prognosis of GC patients

Volcano plot: Red dots refer to the highly expressed genes and green dots refer to the poorly expressed genes. Heatmap: Red refers to the highly expressed genes and green refers to the poorly expressed genes.

Figure 2. LNCRNA AL139002.1 and HAVCR1 expression increased, miR-490-3p expression decreased in GC cell lines. qRT-RCR was applied to detect the relative expression of lncRNA AL139002.1 (a) and miR-490-3p (b) in GC cell lines (MKN-45, AGS, BGC823, SGC7901, MGC803, MKN74) and (GES-1). (c) Western blot of HAVCR1 proteins in GC cell lines (MKN-45, AGS, BGC823, SGC7901, MGC803, MKN74) and (GES-1)

Figure 3. Silencing of lncRNA AL139002.1 regulated proliferation, apoptosis, migration, invasion and EMT of GC cells. LncRNA AL139002.1 was silenced in AGS and MGC-803 cells. (a) qRT-RCR was applied to detect the relative expression of lncRNA AL139002.1 in cells. (b, c) Proliferation and apoptosis assays of cells through CCK-8 assay and flow cytometry. (d) The colony formation of cells results. (e) Migration assay of cells by wound healing analysis (40×). (f) Invasion assay of cells by transwell assay. (g) Western blot of E-cadherin, vimentin and N-cadherin proteins in cells

Figure 4. LncRNA AL139002.1 regulated HAVCR1 by sponging miR‑490‑3p. Relative luciferase activities of lncRNA AL139002.1-wt and lncRNA AL139002.1-mut (a) or HAVCR1-wt and HAVCR1-mut (b) reporter plasmid in AGS and MGC-803 cells co-treated with miR-490-3p mimic. (c) LncRNA AL139002.1 was silenced in AGS and MGC-803 cells. qRT-RCR was conducted to detect the relative expressions of miR-490-3p and HAVCR1. (d) Western blot of HAVCR1 protein in AGS and MGC-803 cells treated with LncRNA AL139002.1 and miR‑490‑3p mimic

Figure 5. LncRNA AL139002.1 regulated the proliferation, apoptosis, migration, invasion and EMT of GC cells by miR‑490‑3p/HAVCR1. Silencing of lncRNA AL139002.1, miR‑490‑3p inhibitor and silencing of HAVCR1 were transfected into AGS and MGC-803 cells. (a, b) Proliferation and apoptosis assays of cells by CCK-8 assay and flow cytometry. (c) The results of the colony formation of cells. (d) Migration assay of cells by wound healing analysis (40×). (e) Invasion assay of cells by transwell assay. (f) Western blot of E-cadherin, vimentin and N-cadherin proteins in AGS and MGC-803 cells

Figure 6. LncRNA AL139002.1/miR‑490‑3p/HAVCR1 regulated GC through MEK/ERK signaling. Silencing of lncRNA AL139002.1, miR‑490‑3p inhibitor and silencing of HAVCR1 were transfected into AGS and MGC-803 cells. (a, b)Western blot of MEK/ERK-related proteins in AGS and MGC-803 cells