miR-939-5p Contributes to the Migration and Invasion of Pancreatic Cancer by Targeting ARHGAP4

Figures & data

Figure 1 ARHGAP4 expression in pancreatic cancer tissues. (A) ARHGAP4 mRNA levels were decreased in pancreatic cancer tissues (n=70) compared with the adjacent-normal tissues (n=70). (B) ARHGAP4 mRNA levels in 70 cases of pancreatic cancer patients (log₂ T/N). (C) ARHGAP4 mRNA expression in five pancreatic cancer cell lines, including BxPC-3, Capan-1, PANC-1, MIA PaCa-2 and SW1990. ***P<0.001 compared with adjacent-normal.

Abbreviations: PC, pancreatic cancer; N, adjacent-normal pancreatic tissues; T, pancreatic cancer tissues.

Figure 2 miR-939-5p targeted ARHGAP4 directly at the 3ʹ-UTR of ARHGAP4 in pancreatic cancer cells. (A) The binding sites of miR-3150a-3p, miR-939-5p and miR-762 on the 3ʹ-UTR of ARHGAP4 were predicted by Targetscan. (B) miR-3150a-3p, miR-939-5p and miR-762 levels in BxPC-3 cells transfected with their corresponding inhibitor or negative control (NC). (C) ARHGAP4 mRNA levels in BxPC-3 cells transfected with miR-3150a-3p, miR-939-5p, or miR-762 inhibitor or NC. (D) The luciferase activity was measured by the Dual-Luciferase Reporter Assay. ***P<0.001 compared with NC.

Table 1 Correlation of the Expression of miR-939-5p with Clinicopathological Parameters in Patients with PC

Clinicopathological Parameters	miR-939-5p Expression		P-value
	Low High
	(n=35, 50%)	(n=35, 50%)
Gender
Male	19 (27.1%)	21 (30.0%)	0.6291
Female	16 (22.9%)	14 (20.0%)
Age (years)
<60	16 (22.9%)	19 (27.1%)	0.4733
≥60	19 (27.1%)	16 (22.9%)
Tumor size (cm)
≤4	20 (28.6%)	16 (22.9%)	0.3388
>4	15 (21.4%)	19 (27.1%)
Pathological stages			0.0072
I	6 (8.6%)	4 (5.7%)
II	20 (28.6%)	8 (11.4%)
III	6 (8.6%)	18 (25.7%)
IV	3 (4.3%)	5 (7.1%)
Tumor site			0.5268
Pancreatic head	14 (20.0%)	10 (14.3%)
Pancreatic body	11 (15.7%)	15 (21.4%)
Pancreatic tail	10 (14.3%)	10 (14.3%)

Note: Differences between groups were done by the Chi-square test.

Figure 3 miR-939-5p expression in pancreatic cancer tissues. (A) miR-939-5p level was increased in pancreatic cancer tissues (n=70) compared with the adjacent-normal tissues (n=70). (B) miR-939-5p levels in 70 cases of pancreatic cancer patients (log₂ T/N). (C) Pearson correlation scatter plots in pancreatic tissues (n=70). (D) Kaplan-Meier curve of pancreatic patients with high or low miR-939-5p levels analyzed by Log-rank (Mantel-Cox) test. ***P<0.001 compared with adjacent-normal.

Abbreviations: PC, pancreatic cancer; N, adjacent-normal pancreatic tissues; T, pancreatic cancer tissues.

Figure 4 miR-939-5p down-regulation inhibited pancreatic cancer cell invasion and migration. (A) ARHGAP4 mRNA expression in five pancreatic cancer cell lines, including BxPC-3, Capan-1, PANC-1, MIA PaCa-2 and SW1990. After BxPC-3 (B, D) and SW1990 (C, E) cells were transfected with miR-939-5p inhibitor, the cell viability, invasion and migration were decreased than those transfected with negative control (NC). ***P<0.001 compared with NC.

Figure 5 miR-939-5p up-regulation promoted pancreatic cancer cell invasion and migration through targeting ARHGAP4. After PANC-1 cells were transfected with miR-939-5p mimic and/or transduced with lentivirus expressing ARHGAP4, miR-939-5p levels (A), ARHGAP4 mRNA levels (B, C) and cell viability (D), migration, and invasion (E, F) were measured. ***P<0.001 compared with NC+blank lentivirus vector (Vector). ^###P<0.001 compared with mimic+Vector.