| Background:MicroRNAs (miRNAs) are a kind of noncoding endogenous single strand ribonucleic acid (RNA) molecules, on average only 21-25 nucleotides long and are widely found in all eukaryotic cells, including plants, animals and viruses. miRNAs match with bases in 3'UTR region, and silence their target genes post-transcriptionally by inhibiting transcription of target mRNAs or degradating them directly, thereby they could regulate the expression of target genes. miRNAs are involved in many complex life processes , such as growth and development, organogenesis, cell proliferation and apoptosis. It is shown that abnormal expression of miRNAs always closely relate to turmorigenesis and development. Hopefully we can find a new strategy to diagnose and treat the tumor on basis of miRNAs. Osteosarcoma is the most common primary bone malignancy in children and adolescents. Half of the osteosarcoma patients are found pulmomary matastases due to early metastases. And the primary factor of death is metastasis. Our lab has established human osteosarcoma cell line SOSP-9607. We then screen two sub-line called F5 and F4 which have different proliferative ability. F5 has a high proliferative and metastatic ability whereas F4 is a rather low proliferative and metastatic sub-line. We found some differentially expressed miRNAs by means of gene chip assay. miR-194 is one of the highly expressed (about 4-fold) miRNAs in F5. This study is to explore the biological function of miR-194 in osteosarcoma cell line SOSP-9607, and supply with theory basis to the further mechanism study as well as prognosis and treatment of osteosarcoma.Objective:To observe the effects of miR-194 mimics on cell proliferation, cycling , apoptosis, aggression and migration of osteosarcoma cell line SOSP-9607.Methods:1. Cultured SOSP-9607 cells were divided into three groups:negtive control group,blank group and experimental group. In the experimental group, osteosarcoma cell line SOSP-9607 are transfected with miR-194 mimics (hsa-miR-194 mimics) in order to enhance the activity of miR-194.2. Draw the growth curves of three groups of cells separately by MTT assay.3. Assay the difference of cell apoptosis and cycling between experimental group and control after transfection with miR-194 mimics by FCM.4. Evaluate the metastatic ability between experimental group and control after transfection with miR-194 mimics by Transwell aggression and migration assay.Results:1. MTT assays show that the proliferative ability of experimental group sharply decreased compared with control(P<0.01).2. FCM show that the apoptosis rate of control group is (3.3±0.19)%, and that of the experimental group is (10.1±0.22)%. The apoptosis rate is obviously increased(P<0.01). Another result of FCM is that the phase G0/G1 cells are increased whereas phase G2/M and S cells decreased remarkably compare with the control group(P<0.01).3. Transwell aggression and migration assay show that the metastatic ability are rather higher than the control group, and there is no significant difference between the two control groups(P<0.01, n=3)..Conclusion:The proliferation, apoptosis, cell cycle, aggression and migration ability of SOSP-9607 are remarkably effected by transfection with miR-194 mimics. However, the accurate mechanism should be studied furtherly. |
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