miR-101a-3p is expressed in a variety of organs and tissues and plays a regulatory role in many diseases,but its role in spinal cord ischemia/reperfusion injury remains unclear.In this study,we established a rat model...miR-101a-3p is expressed in a variety of organs and tissues and plays a regulatory role in many diseases,but its role in spinal cord ischemia/reperfusion injury remains unclear.In this study,we established a rat model of spinal cord ischemia/reperfusion injury by clamping the aortic arch for 14 minutes followed by reperfusion for 24 hours.Results showed that miR-101a-3p expression in L4-L6 spinal cord was greatly decreased,whereas MYCN expression was greatly increased.Dual-luciferase reporter assay results showed that miR-101a-3p targeted MYCN.MYCN immunoreactivity,which was primarily colocalized with neurons in L4-L6 spinal tissue,greatly increased after spinal cord ischemia/reperfusion injury.However,intrathecal injection of an miR-101a-3p mimic within 24 hours before injury decreased MYCN,p53,caspase-9 and interleukin-1βexpression,reduced p53 immunoreactivity,reduced the number of MYCN/NeuN-positive cells and the number of necrotic cells in L4-L6 spinal tissue,and increased Tarlov scores.These findings suggest that the miR-101a-3p mimic improved spinal ischemia/reperfusion injury-induced nerve cell apoptosis and inflammation by inhibiting MYCN and the p53 signaling pathway.Therefore,miR-101a-3p mimic therapy may be a potential treatment option for spinal ischemia/reperfusion injury.展开更多
目的探讨MicroRNA-101(miRNA-101)对上皮性卵巢癌耐药细胞SKOV3/DDP顺铂敏感性的影响及相关作用机制。方法本研究通过细胞转染技术上调SKOV3/DDP细胞内miRNA-101的表达,采用RT-PCR检测miRNA-101及BDNF m RNA的表达情况;CCK-8检测SKOV3/...目的探讨MicroRNA-101(miRNA-101)对上皮性卵巢癌耐药细胞SKOV3/DDP顺铂敏感性的影响及相关作用机制。方法本研究通过细胞转染技术上调SKOV3/DDP细胞内miRNA-101的表达,采用RT-PCR检测miRNA-101及BDNF m RNA的表达情况;CCK-8检测SKOV3/DDP细胞顺铂敏感性及增殖力;Annexin V-FITC/PI流式细胞实验检测细胞凋亡率。结果与阴性对照组比较,miRNA-101转染组细胞miRNA-101表达量及细胞凋亡率增加[(1.000±0.022)vs(5.380±0.246),P<0.05],[(11.020±0.685)%vs(26.158±1.278)%,P<0.05];BDNF m RNA表达量降低[(1.000±0.042)vs(0.389±0.055),P<0.05];顺铂IC50及细胞增殖力均下降[(57.276±1.717)vs(33.176±2.465),P<0.05],[(1.776±0.030)vs(1.642±0.252),P<0.05]。结论 miRNA-101可有效增加SKOV3/DDP细胞对顺铂的药物敏感性,促进细胞凋亡。展开更多
Emerging evidence has indicated that circular RNAs（circRNAs） play pivotal roles in the regulation of cellular processes and are found to be aberrantly expressed in a variety of tumors. However, the clinical role of ...Emerging evidence has indicated that circular RNAs（circRNAs） play pivotal roles in the regulation of cellular processes and are found to be aberrantly expressed in a variety of tumors. However, the clinical role of circ RNAs in bladder cancer（BC） and the molecular mechanisms have yet to be fully understood. In this study, the clinical specimens were obtained and the expression level of a circ RNA BCRC4 was detected by real-time PCR in both BC tissues and cell line. The circular RNA over-expression plasmid was constructed and transfected into BC cells and related cell line. The cell cycles and apoptosis were observed using inverted microscope and flow cytometry. Western blotting was used to compare the relative protein expression of groups with different treatments. It was found that circ RNA BCRC4 expression was lower in BC tissues than in adjacent normal tissues. Furthermore, consequences of forced-expression of BCRC4 promoted apoptosis and inhibited viability of T24T and UMUC3 cells, and up-regulated BCRC4-increased miR-101 level, which suppressed EZH2 expression in both RNA and protein levels. In addition, gambogic acid（GA） is a promising natural anticancer compound for BC therapy, and GA treatment increased the BCRC4 expression in T24T and UMUC3 cells in a dose-dependent manner. Altogether, our findings suggest that BCRC4 functions as a tumor suppressor in BC, and mediates anticancer function, at least in part, by up-regulating the expression of miR-101. Targeting this newly identified circ RNA may help us develop a novel strategy for treating human BC.展开更多
文摘miR-101a-3p is expressed in a variety of organs and tissues and plays a regulatory role in many diseases,but its role in spinal cord ischemia/reperfusion injury remains unclear.In this study,we established a rat model of spinal cord ischemia/reperfusion injury by clamping the aortic arch for 14 minutes followed by reperfusion for 24 hours.Results showed that miR-101a-3p expression in L4-L6 spinal cord was greatly decreased,whereas MYCN expression was greatly increased.Dual-luciferase reporter assay results showed that miR-101a-3p targeted MYCN.MYCN immunoreactivity,which was primarily colocalized with neurons in L4-L6 spinal tissue,greatly increased after spinal cord ischemia/reperfusion injury.However,intrathecal injection of an miR-101a-3p mimic within 24 hours before injury decreased MYCN,p53,caspase-9 and interleukin-1βexpression,reduced p53 immunoreactivity,reduced the number of MYCN/NeuN-positive cells and the number of necrotic cells in L4-L6 spinal tissue,and increased Tarlov scores.These findings suggest that the miR-101a-3p mimic improved spinal ischemia/reperfusion injury-induced nerve cell apoptosis and inflammation by inhibiting MYCN and the p53 signaling pathway.Therefore,miR-101a-3p mimic therapy may be a potential treatment option for spinal ischemia/reperfusion injury.
文摘目的探讨MicroRNA-101(miRNA-101)对上皮性卵巢癌耐药细胞SKOV3/DDP顺铂敏感性的影响及相关作用机制。方法本研究通过细胞转染技术上调SKOV3/DDP细胞内miRNA-101的表达,采用RT-PCR检测miRNA-101及BDNF m RNA的表达情况;CCK-8检测SKOV3/DDP细胞顺铂敏感性及增殖力;Annexin V-FITC/PI流式细胞实验检测细胞凋亡率。结果与阴性对照组比较,miRNA-101转染组细胞miRNA-101表达量及细胞凋亡率增加[(1.000±0.022)vs(5.380±0.246),P<0.05],[(11.020±0.685)%vs(26.158±1.278)%,P<0.05];BDNF m RNA表达量降低[(1.000±0.042)vs(0.389±0.055),P<0.05];顺铂IC50及细胞增殖力均下降[(57.276±1.717)vs(33.176±2.465),P<0.05],[(1.776±0.030)vs(1.642±0.252),P<0.05]。结论 miRNA-101可有效增加SKOV3/DDP细胞对顺铂的药物敏感性,促进细胞凋亡。
文摘Emerging evidence has indicated that circular RNAs（circRNAs） play pivotal roles in the regulation of cellular processes and are found to be aberrantly expressed in a variety of tumors. However, the clinical role of circ RNAs in bladder cancer（BC） and the molecular mechanisms have yet to be fully understood. In this study, the clinical specimens were obtained and the expression level of a circ RNA BCRC4 was detected by real-time PCR in both BC tissues and cell line. The circular RNA over-expression plasmid was constructed and transfected into BC cells and related cell line. The cell cycles and apoptosis were observed using inverted microscope and flow cytometry. Western blotting was used to compare the relative protein expression of groups with different treatments. It was found that circ RNA BCRC4 expression was lower in BC tissues than in adjacent normal tissues. Furthermore, consequences of forced-expression of BCRC4 promoted apoptosis and inhibited viability of T24T and UMUC3 cells, and up-regulated BCRC4-increased miR-101 level, which suppressed EZH2 expression in both RNA and protein levels. In addition, gambogic acid（GA） is a promising natural anticancer compound for BC therapy, and GA treatment increased the BCRC4 expression in T24T and UMUC3 cells in a dose-dependent manner. Altogether, our findings suggest that BCRC4 functions as a tumor suppressor in BC, and mediates anticancer function, at least in part, by up-regulating the expression of miR-101. Targeting this newly identified circ RNA may help us develop a novel strategy for treating human BC.
