| Objective: To explore whether long non-coding RNA(lncRNA)myocyte enhancer factors 2C-antisense RNA 1(MEF2C-AS1)can affect the expression of the SOST gene by acting on myocyte enhancer factors2C(MEF2C)gene,thus affecting the function of osteoblast.Methods:(1)Human osteoblast cell line hFOB1.19 was cultured and transfected with the blank plasmid,over-expressed lncRNA MEF2C-AS1 plasmid as the control group and over-expressed MEF2C-AS1 group.Alkaline phosphatase(ALP)detection kit with p-nitrophenyl phosphate as a substrate was used to detect the changes of ALP activity in the two groups;qRT-PCR and Western blot experiments were performed on the control group and over-expressed MEF2C-AS1 group respectively.The effects of over-expression of MEF2C-AS1 on the mRNA and protein levels of MEF2 C and SOST genes were examined.(2)Human osteoblast cell line hFOB1.19 was cultured and transfected with Smart Silencer negative control and lncRNAMEF2C-AS1 Smart Silencer as a control group and silenced MEF2C-AS1 group.Then,qRT-PCR and Western blot experiments were performed on the control group and silenced MEF2C-AS1 group to detect the effect of silenced MEF2C-AS1 on the mRNA and protein levels of MEF2 C and SOST genes.(3)Human osteoblast cell line hFOB1.19 was cultured,the blank plasmids,over-expressed MEF2C-AS1 plasmids,over-expressed MEF2 C plasmids,over-expressed MEF2C-AS1 and MEF2 C plasmids were transfected respectively as the control group,over-expressed MEF2C-AS1 group,over-expressed MEF2 C group,over-expressed MEF2C-AS1 and MEF2 C group.And qRT-PCR,Western blot experiments were used to detect the mRNA level and protein level of the SOST gene in each group.Results:(1)Compared with the control group,the mRNA expression levels of MEF2 C and SOST in the over-expressed MEF2C-AS1 group were decreased(P <0.05)and protein levels were also decreased(P <0.05),but ALP activity was increased(P <0.05).(2)Compared with the control group,the mRNA expression levels of MEF2 C and SOST in the silenced MEF2C-AS1 group were increased(P <0.05),and protein expression was also increased(P <0.05).(3)The mRNA and protein expression levels of the SOST gene in the control group,over-expressed MEF2C-AS1 group,over-expressed MEF2 C group,over-expressed MEF2C-AS1 and MEF2 C group were different(P <0.05).The mRNA and protein expression levels of the SOST gene in the over-expressed MEF2 C group were increased compared with the control group(P <0.05).At the same time,compared with the over-expressed MEF2C-AS1 group,the mRNA and protein expression level of the SOST gene in the over-expressed MEF2 C and MEF2C-AS1 group were increased(P <0.05).Conclusion: LncRNA MEF2C-AS1 could inhibit the expression of the SOST gene by inhibiting the MEF2 C gene in osteoblasts,and promoting the differentiation and maturation of osteoblasts. |
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