目的探讨阿片类生长因子受体假基因1(OGFRP1)对卵巢癌细胞增殖、侵袭和迁移的影响及其作用机制。方法选取120例卵巢癌患者癌组织及正常卵巢组织,实时荧光定量PCR(RT-qPCR)检测OGFRP1和miR-665表达水平;将A2780细胞分为空白(PBS)组、si-N...目的探讨阿片类生长因子受体假基因1(OGFRP1)对卵巢癌细胞增殖、侵袭和迁移的影响及其作用机制。方法选取120例卵巢癌患者癌组织及正常卵巢组织,实时荧光定量PCR(RT-qPCR)检测OGFRP1和miR-665表达水平;将A2780细胞分为空白(PBS)组、si-NC组、si-OGFRP1组、miR-NC组、miR-665组、si-OGFRP1+anti-miR-NC组、si-OGFRP1+anti-miR-665组。四甲基偶氮唑盐比色法(MTT)检测细胞存活率;Transwell检测细胞迁移和侵袭;双荧光素酶报告实验检测OGFRP1和miR-665的靶向关系。结果与正常卵巢组织相比,卵巢癌组织中OGFRP1表达水平升高(2.75±0.27 vs 1.00±0.09),miR-665表达水平降低(0.51±0.05 vs 1.00±0.08)(均P<0.05)。抑制OGFRP1表达或过表达miR-665后,卵巢癌A2780细胞的活性、细胞迁移与侵袭数均降低(均P<0.05)。双荧光素酶报告实验结果显示,OGFRP1靶向调控miR-665;抑制miR-665和OGFRP1表达后,卵巢癌A2780细胞活性升高,迁移、侵袭细胞数增加(P<0.05)。结论抑制OGFRP1表达可抑制卵巢癌A2780细胞增殖、迁移和侵袭,其机制可能与上调miR-665有关。展开更多
Although numerous miRNAs have been discovered,their functions in the different subcellular organelles have remained obscure.In this study,we found that miR-665 was enriched in the nucleus of cardiomyocytes,and then in...Although numerous miRNAs have been discovered,their functions in the different subcellular organelles have remained obscure.In this study,we found that miR-665 was enriched in the nucleus of cardiomyocytes,and then investigated the underlying role of nuclear miR-665 in heart failure.RNA fluorescence in situ hybridization assays in human heart tissue sections and primary cardiomyocytes showed that miR-665 was localized in the nucleus of cardiomyocytes.Increased expression of nuclear miR-665 was observed not only in the cardiomyocytes isolated from the heart of mice treated in vivo by transverse aortic constriction(TAC),but also in phenylephrine(PE)-treated cultured cardiomyocytes in vitro.To further explore the role of miR-665 in heart failure,a type 9 recombinant adeno-associated virus(rAAV)system was employed to manipulate the expression of miR-665 in mice.Overexpression of miR-665 aggravated TAC-induced cardiac dysfunction,while down-expression of miR-665 showed opposite effects.Bioinformatic prediction and biological validation confirmed that the PTEN(phosphatase and tensin homolog)gene was one of the targets of miR-665 in the nucleus.Furthermore,restoring PTEN expression significantly eliminated the destructive effects of miR-665 over-expression in TAC-induced cardiac dysfunction.Our data showed that nuclear miR-665 aggravates heart failure via inhibiting PTEN expression,which provided a therapeutic approach for heart failure.展开更多
文摘目的探讨阿片类生长因子受体假基因1(OGFRP1)对卵巢癌细胞增殖、侵袭和迁移的影响及其作用机制。方法选取120例卵巢癌患者癌组织及正常卵巢组织,实时荧光定量PCR(RT-qPCR)检测OGFRP1和miR-665表达水平;将A2780细胞分为空白(PBS)组、si-NC组、si-OGFRP1组、miR-NC组、miR-665组、si-OGFRP1+anti-miR-NC组、si-OGFRP1+anti-miR-665组。四甲基偶氮唑盐比色法(MTT)检测细胞存活率;Transwell检测细胞迁移和侵袭;双荧光素酶报告实验检测OGFRP1和miR-665的靶向关系。结果与正常卵巢组织相比,卵巢癌组织中OGFRP1表达水平升高(2.75±0.27 vs 1.00±0.09),miR-665表达水平降低(0.51±0.05 vs 1.00±0.08)(均P<0.05)。抑制OGFRP1表达或过表达miR-665后,卵巢癌A2780细胞的活性、细胞迁移与侵袭数均降低(均P<0.05)。双荧光素酶报告实验结果显示,OGFRP1靶向调控miR-665;抑制miR-665和OGFRP1表达后,卵巢癌A2780细胞活性升高,迁移、侵袭细胞数增加(P<0.05)。结论抑制OGFRP1表达可抑制卵巢癌A2780细胞增殖、迁移和侵袭,其机制可能与上调miR-665有关。
基金supported by grant from the National Natural Science Foundation of China(81822002,81630010,81790624,91839302,31771264 and 31800973)。
文摘Although numerous miRNAs have been discovered,their functions in the different subcellular organelles have remained obscure.In this study,we found that miR-665 was enriched in the nucleus of cardiomyocytes,and then investigated the underlying role of nuclear miR-665 in heart failure.RNA fluorescence in situ hybridization assays in human heart tissue sections and primary cardiomyocytes showed that miR-665 was localized in the nucleus of cardiomyocytes.Increased expression of nuclear miR-665 was observed not only in the cardiomyocytes isolated from the heart of mice treated in vivo by transverse aortic constriction(TAC),but also in phenylephrine(PE)-treated cultured cardiomyocytes in vitro.To further explore the role of miR-665 in heart failure,a type 9 recombinant adeno-associated virus(rAAV)system was employed to manipulate the expression of miR-665 in mice.Overexpression of miR-665 aggravated TAC-induced cardiac dysfunction,while down-expression of miR-665 showed opposite effects.Bioinformatic prediction and biological validation confirmed that the PTEN(phosphatase and tensin homolog)gene was one of the targets of miR-665 in the nucleus.Furthermore,restoring PTEN expression significantly eliminated the destructive effects of miR-665 over-expression in TAC-induced cardiac dysfunction.Our data showed that nuclear miR-665 aggravates heart failure via inhibiting PTEN expression,which provided a therapeutic approach for heart failure.
