ABSTRACT
Objective To investigate the expression of miRNA-182 in hepatocellular carcinoma (HCC) cell lines and tissues, to verify the targeted regulation relationship between miRNA -182 and forkhead box transcription factor 2 (FOXF2) and to explore whether miRNA-182 can promote the invasion, metastasis and angiogenesis of HCC by targeting FOXF2.Methods From January to October 2017, a total of 41 patients with primary HCC admitting to Cancer Hospital Affiliated to Zhengzhou University and receiving operation were enrolled.The expression of miRNA-182 in tumor tissues and normal paracancerous tissues was detected by quantitative real-time fluorescence polymerase chain reaction ( qRT-PCR).The effects of overexpression of miRNR-182 and FOXF2 on invasion and metastasis of HCC were detected by Transwell transfer experiment and the Boyden invasion experiment.The effects of miRNA-182 and FOXF2 on angiogenesis of HCC were determinded by in vitro angiogenesis experiments.The targeted regulation relationship between miRNA -182 and FOXF2 was investigated by dual luciferase reporter gene .The expression of FOXF2 at mRNA and protein level after miRNA-182 and miRNA-182 scramble transfection was detected by qRT -PCR and Western blotting method.T test was performed for statistical analysis.Results The expression in miRNA-182 in HCC tissue was higher than that in normal tissues adjacent to cancer (5.41 ±1.72 vs.1.80 ±0.76), and the difference was statistically significant (t =-5.764, P <0.01).The results of Transwell transfer experiment and the Boyden invasion experiment showed that the number of cells passing through the membrane of the small chamber of miRNA -182 over-expression group was higher than that of miRNA-182 scramble group (85.65 ±4.86 vs.31.43 ±2.41, 55.34 ±5.66 vs. 19.13 ±2.12), the number of cells passing through the membrane of the small chamber of miRNA -182 +FOXF2 over expression group was less than that of simple miRNA -182 over expression group (11.21 ±3.16 vs. 31.43 ±2.41, 8.67 ±2.83 vs.19.13 ±2.12), and the differences were statistically significant (t =15.110, 9.220, 5.443 and 4.410; all P <0.05).The activity of luciferase of wild FOXF2 and miRNA-182 cotransfection group was lower than that of miRNA -182 scramble group (0.43 ±0.10 vs.0.97 ±0.05), and the difference was statistically significant (t =8.042, P =0.012).However the activity of luciferase of mutated FOXF2 and miRNA-182 cotransfection group was higher than that of miRNA -182 scramble group (0.97 ±0.47 vs.1.06 ±0.52), and the difference was not statistically significant (t =0.934, P =0.402).The results of in vitro angiogenesis experiments demonstrated that the number of constituent vessels in miRNA -182 over expression group was higher than that in miRNA-182 scramble group (14.27 ±1.77 vs.5.91 ±1.42), and after FOXF2 over-expressing plasmids cotransfected, the angiogenesis ability restored (1.78 ±0.71 vs.14.27 ±1.77), and the differences were statistically significant (t =6.530 and 4.570, both P <0.05).The results of Western blotting indicated that the expression level of FOXF 2 in miRNA-182 transfection group was lower than that of miRNA-182 scramble group (1.01 ±0.13 vs.4.32 ±0.46), and the difference was statistically significant (t =7.420, P =0.012).The results of qRT-PCR showed that the expression level of FOXF2 mRNA of miRNA-182 transfection group was lower than that in miRNA-182 scramble group (0.591 ±0.284 vs.1.534 ±0.345), and the difference was statistically significant (t =3.465, P =0.014).Conclusion miRNA-182 can promote the invasion and metastasis of HCC cell line by targeting regulation of FOXF 2.
ABSTRACT
Objective To explore the diagnostic value of serum miRNA-182 in colorectal cancer(CRC).Methods A total of 53 CRC patients were enrolled in this study,serum miRNA-182 was detected by quantitative real time polymerase chain reaction(qRT-PCR).The results were compared with those of patients with benign intestinal diseases and healthy controls.Results The content of serum miRNA-182 in CRC patients was significantly higher than those in patients with benign intestinal diseases and healthy group(P0.05).Serum miRNA-182 level in CRC patients after surgery was significantly lower than that before operation(P<0.01),and which was significantly higher in patients with recurrence/metastasis after surgery(P=0.019).Conclusion Serum miRNA-182 level increased significantly in CRC patients,and which was closely related to the disease and malignant progression,serum miRNA-182 could be used as a potential marker for clinical diagnosis and disease evaluation of CRC.