| Background and purposeOsteosarcoma(Osteosarcoma OS)is one of the particularly common malignant bone tumors in adolescence,it is particularly high malignant degree of invasive tumor,and high fatality rate,its characterized by malignant spindle stromal cells can produce bone-like tissue,multiple in the limbs long backbone epiphyseal,one of the most common is the distal femur,clinically found basic is in the middle and late.Although significant advances have been made in clinical treatment for OS,including surgical resection,neoadjuvant or adjuvant chemotherapy and radiotherapy,patients have not improved viability after metastasis or relapse.Long-chain noncoding RNA(Long noncoding RNAs,lnc RNAs)is a novel non-coding RNA with more than 200 nucleotides long,but it does not encode protein functions.To date,there is increasing literature demonstrating that lnc RNA can significantly regulate the pathophysiological properties and phenotypes of OS.They involve a variety of processes,mainly including gene expression,chromatin remodeling,and posttranscriptional processing.This experiment focused on the expression of interfering RNA: LncRNA SNHG12(LncSNHG12)gene by si RNA-LncSNHG12 molecule(si LncSNHG12),the expression of LncSNHG12 gene,and the proliferation and invasion of OS cells under two expressions,which provided beneficial value for clinical diagnosis and treatment of OS.Methods1.Three osteosarcoma cell lines 143 B,MNNG and HOS were purchased from ATCC and the osteoblast cell line h FOB 1.19 cell line was purchased from the Chinese Academy of Sciences(Shanghai,China)culture bank as a reciprocal control assay to CLarify differences in target gene expression.2.The full-length coding sequence of our target gene LncSNHG12 was found in the NCBI gene database,and the most suitable primers for this experiment were made using primer software,and LncSNHG12 primers,LncSNHG12 overexpression plasmid and si LncSNHG12 were synthesized manually by Biocon.3.Purchase 3 Our group has already screened the strongly correlated miRNA of the target gene LncSNHG12 by bioinformatics: miR-195-5p,and prepared its primers with miR-inhabitor in the same way.4.Differential expression of LncSNHG12 in osteosarcoma cell line 143 B versus h FOB 1.19,MNNG,and HOS cell lines was determined using qRT-PCR via primers for LncSNHG12.5.The experiment were divided into three groups: 143 targeted small interference RNA(si LncSNHG12)transfected B Osteosarcoma CL as the silencing group(A);143B Osteosarcoma CL was transfected with the target gene overexpression plasmid(LncSNHG12)as the overexpression group(B),and 143 B Osteosarcoma CL without transfection as the blank control group(C)was cultured routinely;after transfection,the cells were placed in a CO2 incubator at the appropriate temperature in preparation for later experiments.6.The primers for LncSNHG12 and miR-195-5p were used to determine the difference in expression of LncSNHG12 and miR-195-5p in each group using qRT-PCR to assess the correlation,and the CCK8 cell proliferation assay and Transwell cell invasiveness assay were used to observe the difference in cell proliferation and invasiveness in each group to assess the correlation.Results1.A qRT-PCR was performed on 143 B,HOS,and MNNG,as well as on h FOB 1.19 OS CL,Results show: when compared to h FOB 1.19 OS CL,The expression of LncSNHG12 was significantly upregulated in the OS cell lines 143 B,HOS,and MNNG,The results were statistically significant,p<0.05;No significant difference in LncSNHG12 at 143 B CL and MNNG CL,P>0.05;The expression of LncSNHG12 in HOS CL was significantly greater than 143 B CL as well as MNNG CL,P<0.05.2.After qRT-PCR was performed on the silent group,the overexpression group and the blank group,the results showed that the expression level of LncSNHG12 in the silent group was significantly lower than that in the overexpression group and the blank control group,while the expression level in the overexpression group was higher than that in the silent group and the blank control group,i.e.group B > group C > group A,p < 0.05,which was statistically significant;the expression level of miR-195-5p in the silent group was significantly more than that in the The expression level of miR-195-5p in the silent group was significantly higher than that in the blank control group and the overexpression group,while the expression level in the overexpression group was lower than that in the silent group and the blank control group,i.e.group A > group C > group B,p < 0.05,which was statistically significant;the results of proliferation assay and invasion assay of CCK8 cells showed that the proliferation and invasion ability of B and C experimental groups were higher than that of Osteosarcoma cells in A experimental group,while the proliferation and invasion ability of A experimental group and The proliferation and invasion ability of experimental groups A and C were lower than those of experimental group B,i.e.group B > group C > group A,p < 0.05,the difference was statistically significant.Conclusions1.LncSNHG12 was highly expressed in 143 B and MNNG osteosarcoma cell lines,and LncSNHG12 was strongly expressed in HOS osteosarcoma cell line.2.Overexpression of LncSNHG12 resulted in enhanced proliferation and invasion of 143 B osteosarcoma cells,and silencing of LncSNHG12 in the osteosarcoma cell line 143 B reduced the extent of proliferation and invasion of tumour cells.3.In osteosarcoma cells,the expression of LncSNHG12 was negatively correlated with miR-195-5p... |
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