目的:探讨circ-0030042与人第10号染色体缺失的磷酸酶(Phosphatase and tensin homolog deleted on chromosome ten,PTEN)的相互作用关系,并分析其在增生性瘢痕(Hypertrophic scar,HS)患者中对成纤维细胞增殖与迁移的影响及作用机制。方...目的:探讨circ-0030042与人第10号染色体缺失的磷酸酶(Phosphatase and tensin homolog deleted on chromosome ten,PTEN)的相互作用关系,并分析其在增生性瘢痕(Hypertrophic scar,HS)患者中对成纤维细胞增殖与迁移的影响及作用机制。方法:通过circRNA序列和定量聚合酶链反应(PCR技术)检测正常皮肤成纤维细胞(NSFBs)和增生性瘢痕患者成纤维细胞(HSFBs)中circ-0030042的表达。用CCK8检测法检测转染48 h后的HSFBs细胞增殖情况。利用stubRFP-sensGFP-LC3基因转染、流式细胞仪及电子显微镜观察circ-0030042对miR-145/PTEN轴调控VEGF水平的表达。利用生物信息学分析、RNA免疫沉淀、免疫荧光检测等方法,揭示circ-0030042介导HS患者成纤维细胞增殖与迁移的作用机制。结果:circ-0030042在增生性瘢痕中显著上调,过表达时作为VEGF海绵抑制miR-145诱导的成纤维细胞,维持体内稳定性。此外,circ-0030042通过海绵化VEGF水平并阻断其miR-145捕获转录因子(FOXO1)mRNA来影响自噬,而circ-0030042诱导FOXO1的抑制被VEGF水平过表达或circ-0030042结合减少所抵消。过表达circ-0030042对成纤维细胞的增殖抑制与VEGF表达的抑制作用被过表达miR-145部分抵消。结论:干扰circ-0030042通过靶向下调miR-145/PTEN轴进而抑制HSFBs细胞的增殖与迁移,进一步诱导恶性细胞凋亡。展开更多
AIM To determine whether circular RNAs(circ RNAs) are involved in pathological processes of gastric cancer(GC).METHODS Three circ RNAs with differential expression in GC and colorectal cancer were randomly selected fo...AIM To determine whether circular RNAs(circ RNAs) are involved in pathological processes of gastric cancer(GC).METHODS Three circ RNAs with differential expression in GC and colorectal cancer were randomly selected for validation by quantitative reverse transcription-polymerase chain reaction(q RT-PCR), using 20 pairs of gastric tissues and normal tissues. Based on the predicted circ RNAmi RNA network, we then focused on hsa_circ_0000745, which was found to be down-regulated in 20 GC tissues compared with normal tissues. The hsa_circ_0000745 levels were further analyzed by q RT-PCR in 60 GC tissues and paired adjacent non-tumor tissues, as well as 60 plasma samples from GC patients and 60 plasma samples from healthy controls. The associations between the levels of hsa_circ_0000745 and the clinicopathological features of GC patients were statistically assessed. A receiver operating characteristic(ROC) curve was used to evaluate the diagnostic value of hsa_circ_0000745 in GC.RESULTS Hsa_circ_0000745 was down-regulated in GC tissues vs non-tumorous tissues(P < 0.001) and in plasma samples from patients with GC vs healthy controls(P < 0.001). The expression level of hsa_circ_0000745 in GC tissues correlated with tumor differentiation, while the expression level in plasma correlated with tumor-nodemetastasis stage. The area under the ROC curve(AUC) of hsa_circ_0000745 in plasma was 0.683, suggesting good diagnostic value. Plasma hsa_circ_0000745 level combined with carcinoembryogenic antigen(CEA) level increased the AUC to 0.775.CONCLUSION Hsa_circ_0000745 plays an important role in GC and its expression level in plasma in combination with CEA level is a promising diagnostic marker for this malignancy.展开更多
文摘目的:探讨circ-0030042与人第10号染色体缺失的磷酸酶(Phosphatase and tensin homolog deleted on chromosome ten,PTEN)的相互作用关系,并分析其在增生性瘢痕(Hypertrophic scar,HS)患者中对成纤维细胞增殖与迁移的影响及作用机制。方法:通过circRNA序列和定量聚合酶链反应(PCR技术)检测正常皮肤成纤维细胞(NSFBs)和增生性瘢痕患者成纤维细胞(HSFBs)中circ-0030042的表达。用CCK8检测法检测转染48 h后的HSFBs细胞增殖情况。利用stubRFP-sensGFP-LC3基因转染、流式细胞仪及电子显微镜观察circ-0030042对miR-145/PTEN轴调控VEGF水平的表达。利用生物信息学分析、RNA免疫沉淀、免疫荧光检测等方法,揭示circ-0030042介导HS患者成纤维细胞增殖与迁移的作用机制。结果:circ-0030042在增生性瘢痕中显著上调,过表达时作为VEGF海绵抑制miR-145诱导的成纤维细胞,维持体内稳定性。此外,circ-0030042通过海绵化VEGF水平并阻断其miR-145捕获转录因子(FOXO1)mRNA来影响自噬,而circ-0030042诱导FOXO1的抑制被VEGF水平过表达或circ-0030042结合减少所抵消。过表达circ-0030042对成纤维细胞的增殖抑制与VEGF表达的抑制作用被过表达miR-145部分抵消。结论:干扰circ-0030042通过靶向下调miR-145/PTEN轴进而抑制HSFBs细胞的增殖与迁移,进一步诱导恶性细胞凋亡。
基金Supported by National Natural Science Foundation of China,No.81501354
文摘AIM To determine whether circular RNAs(circ RNAs) are involved in pathological processes of gastric cancer(GC).METHODS Three circ RNAs with differential expression in GC and colorectal cancer were randomly selected for validation by quantitative reverse transcription-polymerase chain reaction(q RT-PCR), using 20 pairs of gastric tissues and normal tissues. Based on the predicted circ RNAmi RNA network, we then focused on hsa_circ_0000745, which was found to be down-regulated in 20 GC tissues compared with normal tissues. The hsa_circ_0000745 levels were further analyzed by q RT-PCR in 60 GC tissues and paired adjacent non-tumor tissues, as well as 60 plasma samples from GC patients and 60 plasma samples from healthy controls. The associations between the levels of hsa_circ_0000745 and the clinicopathological features of GC patients were statistically assessed. A receiver operating characteristic(ROC) curve was used to evaluate the diagnostic value of hsa_circ_0000745 in GC.RESULTS Hsa_circ_0000745 was down-regulated in GC tissues vs non-tumorous tissues(P < 0.001) and in plasma samples from patients with GC vs healthy controls(P < 0.001). The expression level of hsa_circ_0000745 in GC tissues correlated with tumor differentiation, while the expression level in plasma correlated with tumor-nodemetastasis stage. The area under the ROC curve(AUC) of hsa_circ_0000745 in plasma was 0.683, suggesting good diagnostic value. Plasma hsa_circ_0000745 level combined with carcinoembryogenic antigen(CEA) level increased the AUC to 0.775.CONCLUSION Hsa_circ_0000745 plays an important role in GC and its expression level in plasma in combination with CEA level is a promising diagnostic marker for this malignancy.
