| Objective :To explore the potential anti-oncogene of bladder cancer,the expression level of mi R-451 was examined,and, analyzed the relation between mi R-451 and bladder carcinoma, s clinical pathological characteristics and metastasis degree of bladder carcinoma cell lines(BC cells). And the ability of adhesion, invasion and metastasis of BC cells with up-regulation of mi R-451 was analyzed by invitro experiments.And to predict and find the target genes of mi R-451. Materials and Methods:30 cases from 2011 /1 to 2014/12 specimens of patients with bladder cancer were collected, The expression level of mi R-451 in 14 cases of bladder cancer tissue was examined using RT-PCR and compared with bladder cancer tissue,pericarcinous tissue and normal tissue,and analyzes the relation between mi R-451 and bladder carcinoma and its clinical pathological characteristics. The expression level of mi R-451 in there bladder carcinoma cell line(T24、5637、 J82) was examined using RT-PCR and compared with each other,and analyzes the relation between mi R-451 and their metastasis degree. The cell model with over expression of mi R-451 was established,the ability of adhesion,invasion and metastasis of BC cells was analyzed with scratch wound model, cellular adhesion experiment and Transwell model,and then, to predict and find the target genes of mi R-451 using bioinformatics softwares like Target Scan,mir Base etc. Results:1. Gene expression test showed that: ① mi R-451 had lower expression in BC tissue than either in PC or in NB tissues(P<0.05). ② the expression of mi R-451 was lower in late-staged(e.g. T3, T4) BC tissue than in early-staged(e.g. T1,T2) BC tissue(P<0.05), and higher in highly differentiated BC tissue than in poorlydifferentiated tissue(P<0.05), which revealed a negative correlation of mi R-451 expression with both T stage and pathological grading. ③ J82, the BC cell strain with lower metastatic potential, had the higher expression of mi R-451, compared with T24 and 5637, showing a negative correlation of mi R-451 expression with the metastatic potential of BC cell strains(P<0.05).2. ① Efficiency test showed that the transfection rates of both mi R-451 mimics and scramble sequence were no less than 90% in each of the three strains, which validated their use in the subsequent experiments. ② It was shown by adhering experiment that the group of BC cells with over expression of mi R-451 had a lower OD(optical density) value than either the negative group or the blank group(P<0.05). No statistical difference, however, was found in cell adhesion between the latter two groups(P>0.05). These results showed that the over expression of mi R-451 could weaken the adhesion capacity of BC cell strains. ③ It was shown by scratch wound test that the group of BC cells with over expression of mi R-451 had the shortest migration distance, compared with the negative group and the blank group, respectively(P<0.05). No statistical difference, however, was found in cell migration between the latter two groups(P>0.05).These results showed that the over expression of mi R-451 could weaken the two-dimensional migration capacity of BC cell strains.④ It was shown by Transwell experiment that the group of BC cells with over expression of mi R-451 had a smaller number of invasive cells than either the negative group or the blank group(P<0.05). No statistical difference, however, was found in number of invasive cells between the latter two groups(P>0.05).These results showed that the over expression of mi R-451 could weaken the invasion capacity of BC cell strains.3. CAB39,MIF,RAB14,ABCB1,etc maybe the target genes of mi R-451 Conclusion:1.The differences of the expression of mi R-451 in bladder cancer tissue, pericarcinous tissue and normal tissue was significant, the expression of mi R-451 maybe closely related to the occurrence, evolution and progress of the bladder cancer.2. There is a strong negative correlation between the expression of mi R-451 in bladder cancer tissue and its clinical pathological characteristics, the mi R-451 may be a new indicator for assessment of the prognosis of patients with bladder cancer.3.The over expression of mi R-451 weakens the two-dimensional adhesion, migration and invasion of bladder carcinoma cell strains.4. RAB14,CAB39,PSMB8,OSR1,MIF,MMP9,etc maybe the target genes of mi R-451.Predicting the target genes with bioinformatics softwares can provide new idear for the research of mi R-451. |
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