MicroRNA-328 enhances cellular motility through posttranscriptional regulation of PTPRJ in human hepatocellular carcinoma

Figures & data

Figure 1 Upregulation of microRNA (miR)-328 and downregulation of PTPRJ messenger RNA (mRNA) in hepatocellular carcinoma (HCC) tissues.

Notes: (A) miR-328 (HCC vs noncancerous: 3.03±0.89 vs 1.95±0.82, P<0.01) expression level was markedly upregulated in HCC tissues compared to adjacent noncancerous tissues. (B) PTPRJ mRNA (HCC vs noncancerous: 2.57±0.91 vs 3.93±0.82, P<0.01) expression level was markedly downregulated in HCC tissues compared to adjacent noncancerous tissues. (C) Upregulation of miR-328 in HCC tissues was significantly negatively correlated with the downregulation of PTPRJ mRNA in HCC tissues (r=−0.362, P=0.01).
Abbreviation: PTPRJ, protein tyrosine phosphatase, receptor type, J.

Table 1 Association of microRNA (miR)-328 and/or PTPRJ mRNA expression with different clinicopathological features of hepatocellular carcinoma patients

Clinicopathological features	Number of cases (%)	miR-328-high (n, %)	P-value	PTPRJ-low (n, %)	P-value	miR-328-high/PTPRJ-low (n, %)	P-value
Age (years)
<50	42 (42.0)	23 (54.8)	NS	22 (52.3)	NS	15 (35.7)	NS
≥50	58 (58.0)	32 (55.2)		30 (51.7)		17 (29.3)
Sex
Male	88 (88.0)	48 (54.5)	NS	46 (52.3)	NS	29 (33.0)	NS
Female	12 (12.0)	7 (58.3)		6 (50.0)		3 (25.0)
HBV infection
Negative	18 (18.0)	10 (55.6)	NS	9 (50.0)	NS	6 (33.3)	NS
Positive	82 (82.0)	45 (54.9)		43 (52.4)		26 (31.7)
Serum AFP level (ng/mL)
<400	25 (25.0)	10 (40.0)	NS	12 (48.0)	NS	6 (24.0)	NS
≥400	75 (75.0)	40 (53.3)		40 (53.3)		26 (34.7)
Venous infiltration
Negative	30 (30.0)	15 (50.0)	NS	15 (50.0)	NS	10 (33.3)	NS
Positive	70 (70.0)	40 (57.1)		37 (52.9)		22 (31.4)
Cirrhosis
Negative	65 (65.0)	35 (53.8)	NS	35 (53.8)	NS	22 (33.8)	NS
Positive	35 (35.0)	20 (57.1)		17 (48.6)		10 (28.6)
Edmondson–Steiner grading
I–II	72 (72.0)	35 (48.6)	0.02	32 (44.4)	0.02	17 (23.6)	0.01
III–IV	28 (28.0)	20 (71.4)		20 (71.4)		15 (53.6)
TNM stage
I–II	70 (70.0)	32 (45.7)	0.02	30 (42.9)	0.02	12 (17.1)	0.01
III–IV	30 (30.0)	23 (76.7)		23 (76.7)		20 (66.7)

Note: “NS” refers to the difference without statistical significance.

Abbreviations: AFP, alpha fetal protein; HBV, hepatitis B virus; TNM, tumor node metastasis; miR, microRNA; mRNA, messenger RNA.

Figure 2 Relative expression of microRNA (miR)-328 in both human hepatocellular carcinoma (HCC) cell lines SMMC-7721 and BEL-7402 transfected with miR-328/normal control (NC) mimics and miR-328/NC inhibitors.

Notes: (A) Expression levels of miR-328 in both SMMC-7721 and BEL-7402 cells transfected with miR-328 mimic were substantially increased, compared to those transfected with NC mimic (*P<0.05). (B) Expression levels of miR-328 in both SMMC-7721 and BEL-7402 cells transfected with miR-328 inhibitor were substantially decreased, compared to those transfected with NC inhibitor (*P<0.05).

Figure 3 MicroRNA (miR)-328 promotes the migration and invasion of hepatocellular carcinoma (HCC) cell lines in vitro.

Notes: (A) Representative images showing the migration and invasion abilities of SMMC-7721 cells transfected with miR-328/normal control (NC) mimics or miR-328/NC inhibitors. (B) Enforced expression of miR-328 significantly promoted cell migration and invasion of both SMMC-7721 and BEL-7402 cells (*P<0.05). Knockdown of miR-328 dramatically retarded cell migration and invasion of both SMMC-7721 and BEL-7402 cells (*P<0.05).

Figure 4 Loss of PTPRJ attenuates the inhibitory effects of knockdown miR-328 on migration and invasion of hepatocellular carcinoma (HCC) cells.

Notes: (A) At 48 hours posttransfection, Western blot analysis revealed that the expression level of PTPRJ protein in cells cotransfected with miR-328 inhibitor and si-PTPRJ was significantly lower than that in cells cotransfected with miR-328 inhibitor and si-con (*P<0.05). (B) Loss of PTPRJ could significantly attenuate the effects of miR-328 inhibitor on cell migration and invasion of both SMMC-7721 and BEL-7402 cells (*P<0.05).
Abbreviations: PTPRJ, protein tyrosine phosphatase, receptor type, J; si-con, matched control siRNA; miRNA, microRNA; si, small intefering.