Zi Chen1, Guohui Zhao, Yunkun Zhang, Yong Ma, Yin Ding, Nanwei Xu. 1. Department of Orthopaedics, Changzhou No.2 People's Hospital Affiliated to Nanjing Medical University, 68th Gehu Middle Rd, Wujing District, Changzhou 213000, Jiangsu, China.
Abstract
PURPOSE: MicroRNAs (miRs) are endogenous, noncoding small RNAs that play a key role in regulating biological and pathological processes. The oncogenic properties of miR-199b-5p have been demonstrated in previous studies but the effect of miR-199b-5p on osteosarcoma (OS) has not yet been clarified. This study aimed to investigate the effect of miR-199b-5p on OS and the relationship between this miR and the pathological parameters and prognosis of OS. METHODS: MiR-199b-5p expression in 57 pairs of OS tissues, corresponding adjacent normal tissues and OS cells was measured by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR).The relationship between miR-199b-5p and the pathological features and prognosis of OS patients was examined. We constructed small interfering (si) RNA to knock down miR-199b-5p expression in OS cell lines MG63 and U2OS. Cell Counting Kit-8 (CCK-8), cell cloning assay and Transwell cell migration and invasion assay were applied for investigating the biological function of miR-199b-5p, respectively. Finally, western blot was used for exploring its underlying mechanism. RESULTS: MiR-199b-5p expression in OS was significantly higher than that of normal tissues. Compared to patients w\sith low expression of miR-199b-5p, patients with high expression level tended to be with younger age, higher incidence of distant metastases and lower overall survival. Compared with interference sequence negative control (si-NC) group, the abilities of proliferation, invasion and metastasis of cells transfected with si-miR-199b-5p were significantly decreased. Western blot analysis indicated that expressions of key proteins related to epithelial to mesenchymal transition (EMT) signaling pathway, including N-cadherin, Vimentin, β-catenin and matrix metalloproteinase-9 (MMP9), were significantly decreased after transfection with si-miR-199b-5p. Furthermore, we found that miR-199b-5p promoted the progression of OS mainly through regulating HER2. CONCLUSIONS: Upregulated miR-199b-5p is significantly related with stage, distant metastasis and poor prognosis of OS. This MiR may promote progression of OS through regulating HER2.
PURPOSE: MicroRNAs (miRs) are endogenous, noncoding small RNAs that play a key role in regulating biological and pathological processes. The oncogenic properties of miR-199b-5p have been demonstrated in previous studies but the effect of miR-199b-5p on osteosarcoma (OS) has not yet been clarified. This study aimed to investigate the effect of miR-199b-5p on OS and the relationship between this miR and the pathological parameters and prognosis of OS. METHODS:MiR-199b-5p expression in 57 pairs of OS tissues, corresponding adjacent normal tissues and OS cells was measured by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR).The relationship between miR-199b-5p and the pathological features and prognosis of OS patients was examined. We constructed small interfering (si) RNA to knock down miR-199b-5p expression in OS cell lines MG63 and U2OS. Cell Counting Kit-8 (CCK-8), cell cloning assay and Transwell cell migration and invasion assay were applied for investigating the biological function of miR-199b-5p, respectively. Finally, western blot was used for exploring its underlying mechanism. RESULTS:MiR-199b-5p expression in OS was significantly higher than that of normal tissues. Compared to patients w\sith low expression of miR-199b-5p, patients with high expression level tended to be with younger age, higher incidence of distant metastases and lower overall survival. Compared with interference sequence negative control (si-NC) group, the abilities of proliferation, invasion and metastasis of cells transfected with si-miR-199b-5p were significantly decreased. Western blot analysis indicated that expressions of key proteins related to epithelial to mesenchymal transition (EMT) signaling pathway, including N-cadherin, Vimentin, β-catenin and matrix metalloproteinase-9 (MMP9), were significantly decreased after transfection with si-miR-199b-5p. Furthermore, we found that miR-199b-5p promoted the progression of OS mainly through regulating HER2. CONCLUSIONS: Upregulated miR-199b-5p is significantly related with stage, distant metastasis and poor prognosis of OS. This MiR may promote progression of OS through regulating HER2.