Expression Of MiRNA-217 In Giant Cell Tumor Of Bone And Its Effect On Autophagy

Objective Giant cell tumor of bone(GCTB)is a borderline bone tumor,and surgical treatment is still the most important treatment.Such tumors are completely cured with difficulty,and the possibility of postoperative recurrence is high.So,the prognosis is poor.Autophagy plays a dual role in the regulation of tumor cells.On the one hand,autophagy can inhibit the occurrence and development of tumors,and on the other hand,autophagy can promote the occurrence and development of tumors.At present,there are few studies on the effect of autophagy on giant cell tumor of bone,and the specific mechanism is still unclear.Many studies have revealed that mi RNAs take on the indispensible role in the occurrence and development of a lot of tumors,and mi RNAs can regulate autophagy.In the process of regulating autophagy,most of them inhibit the occurrence of autophagy,and a small number of micro RNAs can activate autophagy.This study intends to investigate the expression of mi RNA-217 in giant cell tumor of bone and its effect on the autophagy of GCT0404,in order to explore a biomarker that may be used for the early diagnosis,treatment and recurrence of giant cell tumor of bone,and then better provide guidance for clinical diagnosis and treatment.Methods Experiment one: Human osteoblasts h FOB1.19 and GCT0404 cells were cultured in vitro,and the expression levels of mi RNA-217 in osteoblasts h FOB1.19 and GCT0404 were detected using quantitative PCR technique,and finally T-test was performed to compare whether the difference between the two groups was significant.Experiment two: Human GCT0404 cells were cultured in vitro and transfected,and they were divided into NC group,mi RNA-217 mimic group,NC + rapa group,mi RNA-217 mimic + rapa group,inhibitor NC group,mi RNA-217 inhibitor group,inhibitor NC + rapa group,and mi RNA-217 Inhibitor +rapa group,and the expression of m RNA of LC3,Atg5,Atg7,and Beclin1 in the above groups was detected by quantitative PCR;the expression of protein of LC3,Atg5,Atg7,and Beclin1 in the above groups was detected by Western blot;the changes of fluorescence intensity of NAO staining in the above groups were detected by flow cytometry;and the phage formation in the above groups was detected by electron microscopy.Finally,T-test was performed to compare whether the difference between each experimental group and the control group was significant.Results Experiment one: Micro RNA-217 was highly expressed in osteoblasts h FOB1.19 and lower in GCT0404 cells,and there exists the significant difference.Experiment two:(1)The expressions of m RNA of LC3,Atg5,Atg7,and Beclin1 in the cells of the mi RNA-217 mimic group were lower than the NC group,and there exists the significant difference;the expressions of m RNA of LC3,Atg5,Atg7,and Beclin1 in the cells of the NC + rapa,mi RNA-217 mimic + rapa,mi RNA-217 inhibitor,inhibitor NC + rapa groups were higher than the NC group,and there exists the significant difference;the expressions of m RNA of LC3,Atg5,Atg7,and Beclin1 in the NC inhibitor were not significantly different from those in the NC group.(2)The expressions of protein of LC3,Atg5,Atg7,and Beclin1 in the cells of the mi RNA-217 mimic group were lower than the NC group,and there exists the significant difference;the expressions of protein of LC3,Atg5,Atg7,and Beclin1 in the cells of the NC + rapa,mi RNA-217 mimic + rapa,mi RNA-217 inhibitor,inhibitor NC + rapa groups were higher than the NC group,and there exists the significant difference;the expressions of protein of LC3,Atg5,Atg7,and Beclin1 in the NC were not significantly different from those in the NC group.(3)There were significant differences in the fluorescence intensity of NAO between the mi RNA-217 mimic,NC + rapa,mi RNA-217 mimic + rapa,mi RNA-217 inhibitor,inhibitor NC + rapa,and mi RNA-217 inhibitor + rapa groups and the NC group,and the difference in the fluorescence intensity of NAO between the NC inhibitor group was considered not statistically significant.(4)Electron microscopy revealed that mi RNA-217 inhibited autophagosome formation and promoted cellular autophagy when mi RNA-217 expression was down-regulated.Conclusion(1)The expression levels of mi RNA-217 were significantly down-regulated inhuman GCT0404 cells.(2)The expression of autophagy gene-related m RNA and protein was significantly down-regulated in human GCT0404 cells with overexpressing mi RNA-217,and mi RNA-217 inhibited autophagosome formation in human GCT0404 cells,and overexpression of mi RNA-217 was able to inhibit autophagy in GCT0404 cells.