https://orcid.org/0000-0003-0707-0928
Figures & data
Figure 1. Work flowchart of the study.
Figure 2. SLC16A1-AS1 expression and its clinical significance.
(a) Volcano plot demonstrated differentially expressed lncRNAs between normal and OSCC samples. (b) Boxplot indicated lncRNA SLC16A1-AS1 is up-regulated in OSCC samples compared with the normal. (c) Pairwise boxplot also indicated SLC16A1-AS1 high expressed in tumor samples. (d) Analysis of relationship between SLC16A1-AS1 expression and OSCC histologic grades, which revealed high SLC16A1-AS1 accompanied with high tumor grade. (e,f) Kaplan–Meier curve was used for evaluating overall survival time between SLC16A1-AS1 low- and high-expression groups, which revealed SLC16A1-AS1 is a risk factor for OS of OSCC patients and is associated with poor prognosis. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001
Figure 3. The relationship between SLC16A1-AS1 and tumorigenesis-related features.
(a) Composite copy number profiles for low and high SLC16A1-AS1 expression by GISTIC 2.0 amplifications and deletions in OSCC. Chromosomal locations of peaks of significantly recurring focal amplification (red) and deletions (blue) were presented. (b) TMB in patients with low SLC16A1-AS1 expression and high SLC16A1-AS1 expression. (c) Associations between expression of SLC16A1-AS1 and gene mutation statue. The top six significantly related genes were presented. (d) Correlation between mRNA expression-based stemness index (mRNAsi) and SLC16A1-AS1 expression. (e) Correlation of stromal-score, immune-score and ESTIMATE-score with SLC16A1-AS1 expression. (f) SLC16A1-AS1 expression is significantly related to several infiltrating immune cells.
Figure 4. WGCNA identifying the key regulating module and biological functions SLC16A1-AS1 involved in.
(a) Hierarchical clustering dendrogram of genes with dissimilarity based on topological overlap. Modules are the branches of the clustering tree and there were 14 modules identified. (b) Correlation between module eigengenes and SLC16A1-AS1 expression. Each row corresponds to a module eigengene and columns represent a concerning feature. Each cell contains the correlation coefficient and P-value, and magenta module was second most correlated with SLC16A1-AS1. (c) Scatter plot of genes in magenta module. The vertical line represents cutoff of module membership = 0 · 8, and the horizontal line represents cutoff of gene significances for SLC16A1-AS1 = 0.3. The genes met requirements are regarded as SLC16A1-AS1 related key genes. (d) The GO enrichment analysis revealed SLC16A1-AS1 related key genes were involved in multiple aspects of cell proliferation regulation. (e) KEGG pathway analysis revealed SLC16A1-AS1 related key genes are enriched in cell proliferation regulating pathways. (f) PPI network revealed the relationship among these keys genes. The red means a higher MM value and size means the degree in the network.
Figure 5. SLC16A1-AS1 is involved in regulation of OSCC cell proliferation via GSEA and GSVA analysis.
(a–c) GSEA analysis revealed the gene-sets of hallmarks, KEGG pathways and biological processes enriched in SLC16A1-AS1 high expression patients were related to DNA replication, cell cycle process and proliferation, indicating SLC16A1-AS1 takes part in OSCC tumorigenesis. (d) The expression of SLC16A1-AS1 was detected and compared between the high- and low-expression groups. The expression in high-group is much higher than the low. (e–g) GSVA analysis revealed the proliferation scores in high-SLC16A1-AS1 group are significantly higher than the low indicating SLC16A1-AS1 is a positive regulator for OSCC cell proliferation. Data are presented as the means ± SE (error bars). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001
Figure 6. SLC16A1-AS1 silencing inhibits cell proliferation and arrest cell cycle of OSCC cells in vitro.
a. The mRNA expression of commonly used cell proliferation markers between low- and high-SLC16A1-AS1 groups indicating high-SLC16A1-AS1 is associated with high proliferation activity. b. The protein expression of cell proliferation marker PCNA in low- and high-SLC16A1-AS1 group. c. qRT-PCR analysis of the RNA level of SLC16A1-AS1 in cells transfected with NC or SLC16A1-AS1 smart silencer. d,e. Cells transfected with NC or SLC16A1-AS1 smart silencer were examined by CCK8 assays F. Western blot of the protein level of cyclin D1 in cells transfected with NC or SLC16A1-AS1 smart silencer G. Cells with transfection of NC or SLC16A1-AS1 smart silencer were examined by colony formation assay. H. Flow cytometry system analysis was used to detect the phase of the cell cycle in cells transfected with NC or SLC16A1-AS1 smart silencer.
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