In addition, Kaplan-Meier survival curve showed that the overall survival time of patients with lower miR-629 expression was longer than that of patients with higher miR-629 expression levels, suggesting that this upregulation of miR-629 was associated with poor overall survival. proved that this high expression of miR-629 promoted cell proliferation, migration, and invasion of OS. Conclusion All experimental results exhibited that miR-629 as an oncogene promotes the tumor cell growth, migration and invasion of OS, and miR-629 may act as a novel prognostic biomarker and therapeutic target for patients with this malignant tumor. test. Chi-square test was used to evaluate the relationship between miR-629 and clinicopathological characteristics. The relationship between miR-629 and overall survival was estimated by Kaplan-Meier analysis and Cox regression analysis. Results with 0.05 were considered statistically significant. Results Expression of miR-629 in OS Tissues and Cell Lines In order to determine the expression of miR-629 in OS, qRT-PCR was performed in 110 patients. As shown in Physique 1A, miR-629 expression in OS tissue was higher than that in healthy tissues ( 0.001). We then examined the expression of miR-629 in OS cell lines MG63, HOS, SaOS2, U2OS, and the human fetal osteoblastic cell collection hFOB1.19. As shown in Physique 1B, the expression levels of miR-629 in all four OS cell lines were higher than that of human osteoblasts ( 0.001). Open in a separate window Physique 1 The expression of miR-629 in osteosarcoma and normal tissues. (A) miR-629 was significantly upregulated in OS compared to normal tissues (*** 0.001). (B) miR-629 expression in different OS cell lines and human fetal osteoblastic cell collection, the expression levels of miR-629 were higher in all four OS cell lines (*** 0.001). miR-629 Was Correlated with Clinicopathological Characteristics of OS Patients In order to explore the relationship between miR-629 and the clinicopathological characteristics, the OS patients were divided into patients with high miR-629 Rabbit Polyclonal to MAPK1/3 expression group (n = 65) and low miR-629 expression group (n = 45). The relationship between miR-629 expression and various clinicopathological characteristics in OS was shown in Table 1. The results of chi-square analysis indicated that miR-629 overexpression was significantly associated with clinical stage (= 0.031), and distant metastasis (= 0.012). However, miR-629 expression was not correlated with age, gender, tumor size or tumor site ( 0.05). miR-629 Was Correlated with Poor Prognosis in OS Patients The KaplanCMeier method and Log-rank test were used to analyze the relationship between miR-629 expression and the survival time of OS patients, and to explore the prognostic value of miR-629 in OS. The results exhibited that the overall survival time of patients with lower miR-629 expression was longer than that of patients with higher miR-629 expression levels (log-rank = 0.013, Physique 2). Moreover, multivariate Cox regression analysis results indicated miR-629 can be used as an independent prognostic factor in OS (HR = 2.890, 95% CI = 1.126C7.416, = 0.027. Table 2). Table 2 Multivariate Cox Analysis of miR-629 and Clinical Parameters in Relation to Overall Survival = 0.013). miR-629 Regulated Cell Proliferation, Migration, and Invasion in vitro In addition to studying the clinical significance of miR-629 in OS, we further verified whether miR-629 was involved in tumor progression of OS cells by in vitro functional detection. MG63 and U2OS were transfected with miR-629 inhibitor, inhibitor NC, miR-629 mimic, mimic NC. Bethoxazin Transfection efficiency was verified by qRT-PCR for miR-629 expression. Results indicated that miR-629 mimics successfully up-regulated the expression of miR-629, while miR-629 inhibitors down-regulated the expression of miR-629 ( 0.001, Figure 3A). Open in a separate windows Physique 3 Effect of miR-629 on the level of OS cells. Bethoxazin (A) The expression of miR-629 in MG63 and U2OS cells was detected by qRT-PCR after transfection Bethoxazin with miR-629 mimics and inhibitors (** 0.01; *** 0.01; *** 0.001). (D) Transwell analysis was used to detect the effect of miR-629 around the invasion of OS cells, miR-629 mimic significantly promoted cell invasion, and miR-629 inhibitor significantly inhibited cell invasion (*** Bethoxazin 0.001). The effect of miR-629 on cell proliferation ability was examined by CCK-8 assay. The results showed that overexpression of miR-629 promoted cell proliferation, while decreased miR-629 significantly inhibited cell proliferation ( 0.01, Physique 3B). This study also exhibited by Transwell assays that overexpression of miR-629 could promote the migration and invasion ability of OS cells, while the reduction.