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Fig. 3 | Molecular Cancer

Fig. 3

From: Long non-coding RNA LINC00680 functions as a ceRNA to promote esophageal squamous cell carcinoma progression through the miR-423-5p/PAK6 axis

Fig. 3

Transcriptomic analysis reveals that PAK6 is regulated by LINC00680. a KYSE510 cells were subjected to polysome profiling and the resultant fractions were applied to RNA exaction and RT-qPCR analysis to examine the expression of LINC00680. Fractions 1 to 3: Free RNA (unbound with ribosome); Fraction 4: 40S (40S ribosomal subunit); Fractions 5 and 6: 60S (60S ribosomal subunit); Fractions 7 to 9: Monosome; Fractions 10 to 15: Polysome. b-c The subcellular distribution of LINC00680 in KYSE510 (b) and KYSE140 (c) cells was determined by subcellular fractionation followed by RT-qPCR analysis. d KYSE510 and KYSE140 cells transfected with negative control siRNA (si NC) and siRNA specifically targeting LINC00680 (si LINC00680) for three days were subjected to RNA-seq analysis, and gene expression patterns are presented by volcano plot. Blue and red dots represent genes with significant change (P < 0.001, FC > 1.5). e The expression of differentially expressed genes (P < 0.001, FC > 1.5) was represented by heat map. f CeRNA network constituting of LINC00680-miRNAs-mRNAs (genes positively-regulated by LINC00680, n = 68) was shown. Nodes in green, yellow, and light blue represent LINC00680, miRNAs, and mRNAs, respectively. g The flowchart to identify clinically relevant target genes of LINC00680 in ESCC. h The expression of PAK6 in a cohort of esophageal carcinoma (ESCA) samples (n = 182) and normal samples (n = 286) from GEPIA database (http://gepia.cancer-pku.cn/). i The correlation between the expression of PAK6 and prognosis of ESCA patients from Oncolnc database (http://www.oncolnc.org/). j-k KYSE510 and KYSE140 cells were transfected with negative control siRNA (si NC) or two independent siRNAs targeting LINC00680 (si LINC00680 #1 and si LINC00680 #2) followed by RT-qPCR (j) and immunoblotting analysis (k). l The expression of PAK6 in a cohort of ESCC tumor samples (n = 140) and normal samples (n = 140) in house. m The correlation between the expression of PAK6 and prognosis of ESCC patients in house (n = 60). n The correlation between the expression of PAK6 and LINC00680 in ESCC tumor samples in house (n = 140). The experiments were repeated for three times, and representative data is shown with mean ± standard deviation (SD), *P < 0.05, **P < 0.01, ***P < 0.001

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