目的探讨血清外泌体源性circ_0009362对人骨髓间充干细胞(human bone marrow mesenchymalstem cells,hBMSCs)成骨分化的影响及潜在作用机制。方法收集骨质疏松症(osteoporosis,OP)患者的血清并分离外泌体,使用qRT-PCR检测外泌体中circ_0...目的探讨血清外泌体源性circ_0009362对人骨髓间充干细胞(human bone marrow mesenchymalstem cells,hBMSCs)成骨分化的影响及潜在作用机制。方法收集骨质疏松症(osteoporosis,OP)患者的血清并分离外泌体,使用qRT-PCR检测外泌体中circ_0009362的表达水平。诱导hBMSCs成骨并检测细胞中circ_0009362、miR-29b-3p的表达。双荧光素酶报告实验检测circ_0009362、miR-29b-3p的相互调控关系。碱性磷酸酶(ALP)试剂盒检测ALP活性,茜素红(ARS)染色法检测钙沉积。结果与健康对照组相比,OP患者的血清外泌体中circ_0009362表达升高,诱导hBMSCs成骨后circ_0009362的表达降低(均P<0.05)。外泌体能降低hBMSCs中ALP活性与钙沉积百分比,该作用通过分泌circ_0009362实现。敲减外泌体中circ_0009362表达能部分抵消外泌体的作用(均P<0.05)。诱导hBMSCs成骨后miR-29b-3p的表达提高(P<0.05)。circ_0009362与miR-29b-3p存在靶向关系,外泌体通过分泌circ_0009362抑制miR-29b-3p的表达。过表达miR-29b-3p促进hBMSCs中ALP活性与钙沉积百分比,该作用被外泌体部分抵消(均P<0.05)。结论OP患者的血清外泌体通过分泌circ_0009362下调miR-29b-3p的表达来抑制hBMSCs成骨分化。展开更多
To confirm the relationship between Circ_0003855 and EC,we purchased the Human esophageal carcinoma cell line Eca109 and normal human esophageal epithelial cells HEEC,and the expression levels of Circ_0003855,miR-622,...To confirm the relationship between Circ_0003855 and EC,we purchased the Human esophageal carcinoma cell line Eca109 and normal human esophageal epithelial cells HEEC,and the expression levels of Circ_0003855,miR-622,and FLOT1 were detected.The results show that Circ_0003855 and FLOT1 were highly expressed in Eca109 cells,while miR-622 was lowly expressed(p<0.05).Subsequently,Circ_0003855 small interfering RNA(si-Circ_0003855)and its negative control(si-NC)were used to detect changes in cellular biological behaviors.We found that the activity of Eca109 cells was reduced after interfering with the expression of Circ_0003855,and miR-622 expression was elevated,while FLOT1 was decreased(p<0.05).Additionally,si-Circ_0003855 and miR-622 inhibitor sequence(miR-622-inhibition)were co-transfected into cells with miR-622-inhibition alone,and untreated Eca109 cells were used as a control to detect the expression of FLOT1.Co-transfection of si-Circ_0003855 and miR-622-inhibition showed no significant difference in FLOT1 expression compared to the control cells(p>0.05).Synthesizing the results of these experiments above,we believe that interfering with the expression of Circ_0003855 can inhibit the activity of EC cells,and its mechanism is related to miR-622 and FLOT1.展开更多
文摘目的探讨血清外泌体源性circ_0009362对人骨髓间充干细胞(human bone marrow mesenchymalstem cells,hBMSCs)成骨分化的影响及潜在作用机制。方法收集骨质疏松症(osteoporosis,OP)患者的血清并分离外泌体,使用qRT-PCR检测外泌体中circ_0009362的表达水平。诱导hBMSCs成骨并检测细胞中circ_0009362、miR-29b-3p的表达。双荧光素酶报告实验检测circ_0009362、miR-29b-3p的相互调控关系。碱性磷酸酶(ALP)试剂盒检测ALP活性,茜素红(ARS)染色法检测钙沉积。结果与健康对照组相比,OP患者的血清外泌体中circ_0009362表达升高,诱导hBMSCs成骨后circ_0009362的表达降低(均P<0.05)。外泌体能降低hBMSCs中ALP活性与钙沉积百分比,该作用通过分泌circ_0009362实现。敲减外泌体中circ_0009362表达能部分抵消外泌体的作用(均P<0.05)。诱导hBMSCs成骨后miR-29b-3p的表达提高(P<0.05)。circ_0009362与miR-29b-3p存在靶向关系,外泌体通过分泌circ_0009362抑制miR-29b-3p的表达。过表达miR-29b-3p促进hBMSCs中ALP活性与钙沉积百分比,该作用被外泌体部分抵消(均P<0.05)。结论OP患者的血清外泌体通过分泌circ_0009362下调miR-29b-3p的表达来抑制hBMSCs成骨分化。
文摘To confirm the relationship between Circ_0003855 and EC,we purchased the Human esophageal carcinoma cell line Eca109 and normal human esophageal epithelial cells HEEC,and the expression levels of Circ_0003855,miR-622,and FLOT1 were detected.The results show that Circ_0003855 and FLOT1 were highly expressed in Eca109 cells,while miR-622 was lowly expressed(p<0.05).Subsequently,Circ_0003855 small interfering RNA(si-Circ_0003855)and its negative control(si-NC)were used to detect changes in cellular biological behaviors.We found that the activity of Eca109 cells was reduced after interfering with the expression of Circ_0003855,and miR-622 expression was elevated,while FLOT1 was decreased(p<0.05).Additionally,si-Circ_0003855 and miR-622 inhibitor sequence(miR-622-inhibition)were co-transfected into cells with miR-622-inhibition alone,and untreated Eca109 cells were used as a control to detect the expression of FLOT1.Co-transfection of si-Circ_0003855 and miR-622-inhibition showed no significant difference in FLOT1 expression compared to the control cells(p>0.05).Synthesizing the results of these experiments above,we believe that interfering with the expression of Circ_0003855 can inhibit the activity of EC cells,and its mechanism is related to miR-622 and FLOT1.
