MiR-193a-3p inhibits pancreatic ductal adenocarcinoma cell proliferation by targeting CCND1

Figures & data

Figure 1 MiR-193a-3p expression in PDAC. (A) MiR-193a-3p expression was down-regulated in PDAC tissue in the online databases. (B) MiR-193a-3p expression in PANC-1 and BxPC-3 cells was lower than that in Hpde6-C7 cells. (C) MiR-193a-3p expression in PDAC tissues was lower than in para-carcinoma tissues. (D) Forest plot combining the GEO datasets with our qRT-PCR data showed down-regulated expression of miR-193a-3p in PDAC tissue.

Abbreviations: PDAC, pancreatic ductal adenocarcinoma; GEO, Gene Expression Omnibus.

Figure 2 Effect of miR-193a-3p overexpression on PDAC cell proliferation. (A) The proliferation capacity of BxPC-3 cells was significantly decreased 3 d(P=0.001), 4 d(P=0.022), and 5 d(P=0.002) after transfection. (B) The multiplication ability of Panc-1 cells was significantly reduced 2 d(P=0.006), 3 d(P=0.001), 4 d(P=0.011), and 5 d(P=0.000) after transfection. (*P<0.05, **P<0.01, ***P<0.001).

Abbreviations: NC, negative control; OE, the overexpression of miR-193a-3p.

Figure 3 The cycle distribution of PDAC cells after overexpression of miR-193a-3p. (A and B) Cycle distribution of the BxPC-3 cell line and Panc-1 cell line. (C) Statistical analysis of the cell cycle distribution of PDAC.

Abbreviations: NC, negative control; OE, the overexpression of miR-193a-3p.

Figure 4 Effect of miR-193a-3p overexpression on the apoptosis of PDAC cells. (A) The result of detecting apoptosis after the overexpression of miR-193a-3p. (B) Apoptosis rates of the two cell lines in the experimental groups were compared with those in the negative control groups.

Abbreviations: NC, negative control; OE, the overexpression of miR-193a-3p.

Table 1 GO analysis of miR-193a-3p target genes

Term	Type	Count	P-value	FDR
Proteasome-mediated ubiquitin-dependent protein catabolic process	Biological process	9	6.71E-04	1.058
Cell migration	Biological process	8	0.003	4.356
Negative regulation of cell proliferation	Biological process	11	0.004	5.490
Regulation of axon extension involved in axon guidance	Biological process	3	0.004	6.427
Regulation of synaptic transmission, GABAergic	Biological process	3	0.008	11.436
Retrograde transport, endosome to Golgi	Biological process	5	0.010	14.316
Branchiomotor neuron axon guidance	Biological process	3	0.010	14.320
Semaphorin receptor complex	Cellular component	3	0.009	10.941
Transcriptional activator activity, RNA polymerase II core promoter proximal region sequence-specific binding	Molecular function	10	0.007	8.270

Table 2 KEGG pathway analysis of miR-193a-3p target genes

Term	Genes	P-value	FDR
MicroRNAs in cancer	PRKCA, IRS2, BMPR2, IGF2BP1, BCL2L2, MMP16, PTEN, TGFB2, CCND1, KRAS, CRKL, SOS2, PDGFRA, PLAU	9.95E-08	0.000
FoxO signaling pathway	USP7, IGF1R, CCND1, IRS2, KRAS, SOS2, MAPK8, FOXO4, PTEN, TGFB2	1.23E-04	0.152
Pathways in cancer	TRAF1, PRKCA, RUNX1T1, ARHGEF12, PTEN, TGFB2, IGF1R, CCND1, KRAS, CRKL, ADCY9, GNB1, SOS2, PDGFRA, LAMC2, MAPK8, LAMC1	1.74E-04	0.216
Glioma	PRKCA, IGF1R, CCND1, KRAS, SOS2, PDGFRA, PTEN	3.62E-04	0.447
GnRH signaling pathway	KRAS, ADCY9, MAP3K3, MAP3K1, SOS2, MAPK8, MMP14	0.0013	1.642
Focal adhesion	PRKCA, IGF1R, CCND1, CRKL, SOS2, PDGFRA, MAPK8, LAMC2, LAMC1, PTEN	0.0033	4.042
Hepatitis B	PRKCA, CCND1, YWHAZ, KRAS, MAP3K1, MAPK8, PTEN, TGFB2	0.0048	5.807
Transcriptional misregulation in cancer	TRAF1, IGF1R, KMT2A, RUNX1T1, ETV1, SIX4, PLAU, MYCN	0.0077	9.151
Proteoglycans in cancer	PRKCA, IGF1R, CCND1, KRAS, SOS2, RPS6KB2, ARHGEF12, PLAU, TGFB2	0.0079	9.323
ErbB signaling pathway	PRKCA, CRKL, KRAS, SOS2, RPS6KB2, MAPK8	0.009	10.570
Prostate cancer	IGF1R, CCND1, KRAS, SOS2, PDGFRA, PTEN	0.009	10.570
AMPK signaling pathway	IGF1R, CCND1, IRS2, PPP2R5C, ACACA, RPS6KB2, CAB39	0.009	10.633
Acute myeloid leukemia	CCND1, KRAS, SOS2, RUNX1T1, RPS6KB2	0.009	10.703
Axon guidance	PLXNA4, KRAS, SEMA4G, PLXNA1, UNC5B, PLXNA2, ARHGEF12	0.0098	11.428

Figure 5 Bubble Chart of the results of the GO and KEGG analysis. Pathway visualization of the results of the GO and KEGG analysis of the miR-193a-3p target genes, which was performed in ImageGP (https://www.ehbio.com/ImageGP/).

Abbreviations: BP, biological process; CC, cellular component; MF, molecular function; GO, gene ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Figure 6 Protein-protein interaction network of miR-193a-3p target genes.

Figure 7 The expression of CCND1 in PDAC. (A) CCND1 expression in FFPE PDAC tissues and para-carcinoma tissues. (B) Forest plot for the miR-193a-3p expression in PDAC.

Abbreviation: FFPE, formalin-fixed paraffin-embedded.

Figure 8 The relationship between miR-193a-3p and CCND1. (A) Base-complementing sequences of miR-193a-3p and CCND1. (B) Relative luciferase activity in cells with CCND1 3ʹUTR+miR-193a-3p was reduced compared to cells with CCND1 3ʹUTR-NC+miR-193a-3p. The relative luciferase activity in the group of positive reference miRNA was lower than in the group of positive reference miRNA NC. (C) Negative correlation between miR-193a-3p and CCND1 in FFPE PDAC tissue. (D) Reduced expression of CCND1 in the two cell lines after the overexpression of miR-193a-3p.

Abbreviations: NC, negative control; OE, the overexpression of miR-193a-3p; FFPE, formalin-fixed paraffin-embedded.