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Objective: miRNA-194 were knocked down or overexpressed in Osteosarcoma Cell Line SOSP-9607 to investigate its effect on Metastasis of human Osteosarcoma which to provide the foundation of theory of miRNA-194 as a new target of osteosarcoma therapy. Methods: We transfected human osteosarcoma cell lines SOSP-9607 by using recombinant lentiviruses and divided the cells into three groups with different levels of miRNA-194. Then the stable transfected cells were used in transwell and wound healing assay. Results:1) The transfection and selection of miRNA-194 using recombinant lentiviral expression vector was finished correctly and different levels of miRNA-194 were got sucessfully. 2) Results in the transwell migration and invasion assay showed significantly lower numbers of OE SOSP-9607 cells (28.60± 4.36) and (21.25± 6.42) and significantly higher numbers of KD SOSP-9607 cells (132.60± 15.64; 115.76± 11.38) compared with other groups (p<0.05). The wound healing assay showed that cells in OE groups exhibited a significant decrease in migration rate compared to the other three groups(p<0.05). Conclusions: miRNA-194 could inhibit metastasis of Osteosarcoma Cell Line SOSP-9607. MiRNA-194 could be further explored as a potential target in Osteosarcoma diagnosis and therapy. [ABSTRACT FROM AUTHOR]

Objective: To forecast and verify the target genes of miRNA-194 in human osteosarcoma and establish some theoretical basis for osteosarcoma treatment and element experience of miRNA-194 for further research. Methods: Potential target gene of miRNA-194 was predicted using bioinformatics and were verified in gene and protein levels. Moreover, luciferase reporter assay was used to confirm whether CDH2 was direct targets of miRNA-194. Results: Potential target genes of miRNA-194 were predicted by using bioinformatics. Moreover, luciferase activity of the wild-type PTEN-3'-UTR reporter was significantly suppressed in miRNA-194up-regulation group compared with other groups in SOSP-9607 cells(P<0.01) while down-regulation group could significantly increase it(P<0.01). At the same time, western blot analysis revealed that miRNA-194 expression could affect the expression of CDH2 protein while there was no significant difference on m RNA level(P<0.05). Conclusion: CDH2, one target gene of miRNA-194, was predicted and verified, which provided the theoretical basis for the next study of miRNA-194 in the osteosarcoma. [ABSTRACT FROM AUTHOR]

Objective: To explore and describe the relationship of miRNA-192 function and clinicopathological features in human osteosarcoma and analyze the relationship between miRNA-192 and mortality, which support some theoretical basis for osteosarcoma treatment and element experience for further research. Methods: Real-time quantitative PCR analysis were used in 50 clinical osteosarcoma samples which were followed up. Then the relationship of miRNA-192 function and clinicopathological features(like sex,age) in human osteosarcoma were explored. Results: We found that there were statistically significant relationships between miRNA-192 expression and some clinicopathological features of osteosarcoma, including age(P = 0.028), clinical stage(P = 0.007), distant metastasis(P = 0.013) and patient mortality(P=0.001). Conclusion: In clinical specimens, miRNA-192 expression levels were correlated with some clinicopathological features of osteosarcoma, including age, clinical stage, distant metastasis and patient mortality, which provided the theoretical basis for the next study especially the biological therapy of miRNA-192 in the osteosarcoma. [ABSTRACT FROM AUTHOR]

Objective: We examined the effects of miRNA-194 on the capacities of tumor formation and pulmonary metastasis of osteosarcoma in nude mice by using recombinant lentiviruses. All these wouLd provide the foundation of theory to explore microRNA- 194 as a new target of osteosarcoma therapy. Methods: we transfected human osteosarcoma cell lines U2-OS by using recombinant lentiviruses and divided the mice into three groups. Then we put the cells into the mice to built the metastasis model of rude mice. 5 weeks later the mice were killed to compare the size and weight of tumor in leg and lung. Results: 1) The size and weight of miRNA- 194-down were obviously higher than that in other groups (P<0.05). 2) The lung of rude mice of miRNA-194-up cell were obviously better than that in other groups(P<0.05). 3) It were certified that the tumor were osteosarcoma and metastatic tumor. Conclusion: In vivo, miRNA-194 couLd significantly inhibit the capacities of tumor formation and pulmonary metastasis which declare that miRNA-194 may play the role of tumor suppressor genes in osteosarcoma, which provide a good foundation for the study of miRNA-194. [ABSTRACT FROM AUTHOR]

Objective: Use microRNA microarray chip and real time quantitative PCR method to analysis the relative expression levels of miR-15a-5p and miR-16-5p, then compare the difference between osteosarcoma paraffin block samples and peritumoral tissue. Methods: 34 cases of osteosarcoma paraffin block samples were chosen, microRNAs microarray chip and real time quantitative PCR method was used to measure the relative expression levels of miR-15a-5p and miR-16-5p. Results: microRNAs microarray chip results showed that the expression level ofmiR-15a-5p (miR-16-5p) in tumor tissue was 1.79 (1.62) times lower than the adjacent tissue. Real time quantitative PCR results showed that compare with tumor adjaccnt tissues, the expression level of miR-15a-5p and miR-16-5p was low in osteosarcoma paraffin block samples. The results were considered significant through statistical analysis (P＜0.05). Further analysis results showed that the expression level of miR-15a-5p (miR-16-5p) in tumor tissue was 3.14 (5.65) times lower than the adjacent tissue. Conclusions: miR-15a-5p and miR-16-5p was low expressed in osteosarcoma cells, which suggest that these two microRNAs could be act as oncogene. [ABSTRACT FROM AUTHOR]

目的探讨微小RNA-432(miR-432)对骨肉瘤细胞增殖、迁移和侵袭的影响及其作用机制。方法采用qRT-PCR技术检测人骨肉瘤细胞MG-63、人成骨细胞h FOB1.19中miR-432表达。将对数生长期MG-63细胞随机分为三组,分别转染miRNA对照质粒(miR-NC组)、miR-432抑制质粒(miR-432 inhibitor组)及miR-432过表达质粒(miR-432组),转染48 h时进行接种培养,分别检测细胞增殖(吸光度值)、迁移和侵袭情况。利用miRWalk和miRanda生物信息学软件预测miR-432靶基因可能为致癌基因CX3CL1,分别构建含有miR-432结合位点的CX3CL1基因3’-UTR区野生型(CX3CL1-wt)和突变型(CX3CL1-mut)荧光素酶报告载体。将MG-63细胞分别共转染CX3CL1-wt或CX3CL1-mut报告载体和miR-432载体(观察组),并设置共转染miR-NC载体作为对照组。转染48 h时,采用双荧光素酶试剂盒检测相对荧光素酶活性。结果 MG-63细胞中miR-432相对表达量低于h FOB1.19细胞(P<0.01)。miR-NC组、miR-432 inhibitor组、miR-432组细胞中miR-432相对表达量分别为1.00±0.01、0.47±0.03、3.62±0.03,组间比较P均<0.01。miR-432组培养24、36、48 h的细胞吸光度值均明显低于miR-NC组、miR-432 inhibitor组,miR-432 inhibitor组均明显高于miR-NC组。miR-432 inhibitor组及miR-432组细胞相对迁移率分别为(122±7)%、(65±4)%,相对侵袭率分别为(189±25)%、(49±11)%;上述指标组间比较P均<0.05。CX3CL1-wt和miR-432共转染MG-63细胞后,观察组相对荧光素酶活性低于对照组(P<0.05);CX3CL1-mut和miR-432共转染细胞后,对照组和观察组相对荧光素酶活性分别为1.00±0.11、0.93±0.13,两组比较P>0.05。结论 miR-432过表达可抑制骨肉瘤细胞增殖、迁移和侵袭,靶向调控致癌基因CX3CL1可能是其作用机制。 [ABSTRACT FROM AUTHOR]