Aim: To reveal the exonic and 3’UTR sequences of KRAS, TP53, APC, BRAF, PIK3CA genes in sporadic colorectal tumors and to investigate the clinical relevance of 3’UTR variations in miRNA profiles. Methods: In the stu...Aim: To reveal the exonic and 3’UTR sequences of KRAS, TP53, APC, BRAF, PIK3CA genes in sporadic colorectal tumors and to investigate the clinical relevance of 3’UTR variations in miRNA profiles. Methods: In the study, the exonic and 3’UTR sequences of five genes in 12 sporadic colorectal tumors were extracted by next generation sequencing. In tumors with variation in the 3’UTR region, the changes caused by the variation in the miRNA binding profile were detected. The expression profile of these miRNAs in colorectal and other solid tumors compared to normal tissue was determined. Pathway analysis was performed to determine which signaling pathways miRNAs affect. Results: Case-10 in our study was wild type KRAS and received cetuximab treatment and developed drug resistance. In this case, it was concluded that the expression of KRAS increased and tumorigenesis progressed due to miRNAs that do not bind to this region due to variations in the 3’UTR region. Among these miRNAs, hsa-miR-124-3p was found to have decreased expression in colorectal tumors and to be associated with the ECM-receptor interaction pathway. Conclusion: Variations in the 3’UTR regions of genes critical in the process of carsinogenesis are associated with drug resistance and the process of tumorigenesis.展开更多
Objective： To explore the effects of acupuncture at Baihui （GV 20） and Zusanli （ST 36） on the peripheral serum expression of microRNA 124 （miRNA 124）, laminin and integrinβ 1 in rats with cerebral ischemia rep...Objective： To explore the effects of acupuncture at Baihui （GV 20） and Zusanli （ST 36） on the peripheral serum expression of microRNA 124 （miRNA 124）, laminin and integrinβ 1 in rats with cerebral ischemia reperfusion injury （CIRI）. Methods： Seventy-two healthy male Sprague-Dawley rats were randomized into a model group, an acupuncture group, and a sham-operated group using a random digits table, with 24 rats per group. Each group was further randomly divided into 1-, 3-, 5-, and 7-day subgroups based on the reperfusion time according to a random digits table, with 6 rats in each subgroup. In the model and acupuncture groups, CIRI was induced using the thread occlusion method. Electroacupuncture stimulation was applied daily to GV 20 and left ST 36 for 20 min at the indicated time points after successful operations. Serum was sampled for detecting laminin and integrin β 1 protein via enzyme-linked immunosorbent assay, and serum miRNA 124 was examined using quantitative polymerase chain reaction. Results： The serum level of miRNA 124 in the cerebral ischemia rats increased significantly, and the peak expression of miRNA 124 in both the model and acupuncture groups occurred at 3 days. The expression of miRNA 124 in the acupuncture group was higher than in the model group at the same time point （5.96±0.01 vs. 3.11 ±0.04, P〈0.05）. Laminin expression in serum from the cerebral ischemia group was higher than that in the sham-operated group. Compared with the model group, the level of laminin in the serum of the acupuncture group was significantly lower at each time point, especially at the 3-day, and 7-day time points （589.12± 3.57 vs. 793.05± 5.28, and 600.53± 3.05 vs. 899.06± 5.74, P〈0.05）. The level of integrin 131 in the serum from the acupuncture group was lower than that in the model group particularly at the 3-day and Z-day time points （208.66± 0.95 vs. 280.83± 1.77, and 212.36±0.95 vs. 316.77±2.42, P〈0.05）. Additionally, the model group and the acupuncture group showed dual peaks of integrin β 1 and laminin expression at 3-day and 7-day. Conclusions： Acupuncture at GV 20 and ST 36 in rats alleviated CIRI and was associated with upregulated expression of miRNA 124 and with downregulated expression of integrin β 1 and laminin in peripheral serum. These changes may represent one of the mechanisms underlying acupuncture＇s attenuation of CIRI.展开更多
目的探究微小RNA-124(miR-124)通过特异性β1糖蛋白(Sp1)控制人血管平滑肌细胞(HVSMC)表型转换影响颅内动脉瘤(IA)发展的机制。方法HVSMC进行原代培养,HVSMC在5%胎牛血清的生长培养基SmGM-2(Lonza)中于37℃在加湿的5%CO_(2)培养箱中培...目的探究微小RNA-124(miR-124)通过特异性β1糖蛋白(Sp1)控制人血管平滑肌细胞(HVSMC)表型转换影响颅内动脉瘤(IA)发展的机制。方法HVSMC进行原代培养,HVSMC在5%胎牛血清的生长培养基SmGM-2(Lonza)中于37℃在加湿的5%CO_(2)培养箱中培养。根据研究方案将HVSMC细胞分为对照组、miR-124模拟转染组和miR-124-pCMV-Sp1Mut共转染组。通过定量RT-PCR分析HVSMC在血小板源生长因子(PDGF-BB)刺激后miR-124表达。通过BrdU检测和刮擦试验检测HVSMC的增殖迁移。通过荧光素酶报告基因测定检测miR-124与Sp1的靶向关系。通过蛋白印迹分析HVSMC标记基因和Sp1的蛋白表达。结果与0 h相比,PDGF-BB(20 ng/mL)诱导后第6、12、24 h HVSMC中miR-124表达降低,差异有统计学意义(P<0.05)。miR-124模拟转染组细胞增殖和细胞迁移能力较对照组增强,差异均有统计学意义(P<0.05)。miR-124模拟转染组与Sp1-WT3'-UTR融合的荧光素酶活性较对照组降低,差异有统计学意义(P<0.05),而miR-124模拟转染组与Sp1-MUT的3'-UTR融合的荧光素酶活性与对照组相比,差异无统计学意义(P>0.05)。miR-124模拟转染组α-SMA、SM22a、calponin和MYH11蛋白表达较对照组升高,miR-124-pCMV-Sp1Mut共转染组α-SMA、SM22a、calponin和MYH11蛋白表达较miR-124模拟转染组降低,差异均有统计学意义(P<0.05)。miR-124模拟转染组Sp1蛋白表达较对照组降低,miR-124-pCMV-Sp1Mut共转染组Sp1蛋白表达较miR-124模拟转染组升高,差异均有统计学意义(P<0.05)。结论miR-124通过Sp1调节HVSMC的表型并显著抑制IA发展,miR-124对Sp1表达的调节可能为miR-124的功能脑血管疾病的分子机制提供新的见解。展开更多
文摘Aim: To reveal the exonic and 3’UTR sequences of KRAS, TP53, APC, BRAF, PIK3CA genes in sporadic colorectal tumors and to investigate the clinical relevance of 3’UTR variations in miRNA profiles. Methods: In the study, the exonic and 3’UTR sequences of five genes in 12 sporadic colorectal tumors were extracted by next generation sequencing. In tumors with variation in the 3’UTR region, the changes caused by the variation in the miRNA binding profile were detected. The expression profile of these miRNAs in colorectal and other solid tumors compared to normal tissue was determined. Pathway analysis was performed to determine which signaling pathways miRNAs affect. Results: Case-10 in our study was wild type KRAS and received cetuximab treatment and developed drug resistance. In this case, it was concluded that the expression of KRAS increased and tumorigenesis progressed due to miRNAs that do not bind to this region due to variations in the 3’UTR region. Among these miRNAs, hsa-miR-124-3p was found to have decreased expression in colorectal tumors and to be associated with the ECM-receptor interaction pathway. Conclusion: Variations in the 3’UTR regions of genes critical in the process of carsinogenesis are associated with drug resistance and the process of tumorigenesis.
基金Supported by the National Natural Science Foundation of China(No.81273850)
文摘Objective： To explore the effects of acupuncture at Baihui （GV 20） and Zusanli （ST 36） on the peripheral serum expression of microRNA 124 （miRNA 124）, laminin and integrinβ 1 in rats with cerebral ischemia reperfusion injury （CIRI）. Methods： Seventy-two healthy male Sprague-Dawley rats were randomized into a model group, an acupuncture group, and a sham-operated group using a random digits table, with 24 rats per group. Each group was further randomly divided into 1-, 3-, 5-, and 7-day subgroups based on the reperfusion time according to a random digits table, with 6 rats in each subgroup. In the model and acupuncture groups, CIRI was induced using the thread occlusion method. Electroacupuncture stimulation was applied daily to GV 20 and left ST 36 for 20 min at the indicated time points after successful operations. Serum was sampled for detecting laminin and integrin β 1 protein via enzyme-linked immunosorbent assay, and serum miRNA 124 was examined using quantitative polymerase chain reaction. Results： The serum level of miRNA 124 in the cerebral ischemia rats increased significantly, and the peak expression of miRNA 124 in both the model and acupuncture groups occurred at 3 days. The expression of miRNA 124 in the acupuncture group was higher than in the model group at the same time point （5.96±0.01 vs. 3.11 ±0.04, P〈0.05）. Laminin expression in serum from the cerebral ischemia group was higher than that in the sham-operated group. Compared with the model group, the level of laminin in the serum of the acupuncture group was significantly lower at each time point, especially at the 3-day, and 7-day time points （589.12± 3.57 vs. 793.05± 5.28, and 600.53± 3.05 vs. 899.06± 5.74, P〈0.05）. The level of integrin 131 in the serum from the acupuncture group was lower than that in the model group particularly at the 3-day and Z-day time points （208.66± 0.95 vs. 280.83± 1.77, and 212.36±0.95 vs. 316.77±2.42, P〈0.05）. Additionally, the model group and the acupuncture group showed dual peaks of integrin β 1 and laminin expression at 3-day and 7-day. Conclusions： Acupuncture at GV 20 and ST 36 in rats alleviated CIRI and was associated with upregulated expression of miRNA 124 and with downregulated expression of integrin β 1 and laminin in peripheral serum. These changes may represent one of the mechanisms underlying acupuncture＇s attenuation of CIRI.
文摘目的探究微小RNA-124(miR-124)通过特异性β1糖蛋白(Sp1)控制人血管平滑肌细胞(HVSMC)表型转换影响颅内动脉瘤(IA)发展的机制。方法HVSMC进行原代培养,HVSMC在5%胎牛血清的生长培养基SmGM-2(Lonza)中于37℃在加湿的5%CO_(2)培养箱中培养。根据研究方案将HVSMC细胞分为对照组、miR-124模拟转染组和miR-124-pCMV-Sp1Mut共转染组。通过定量RT-PCR分析HVSMC在血小板源生长因子(PDGF-BB)刺激后miR-124表达。通过BrdU检测和刮擦试验检测HVSMC的增殖迁移。通过荧光素酶报告基因测定检测miR-124与Sp1的靶向关系。通过蛋白印迹分析HVSMC标记基因和Sp1的蛋白表达。结果与0 h相比,PDGF-BB(20 ng/mL)诱导后第6、12、24 h HVSMC中miR-124表达降低,差异有统计学意义(P<0.05)。miR-124模拟转染组细胞增殖和细胞迁移能力较对照组增强,差异均有统计学意义(P<0.05)。miR-124模拟转染组与Sp1-WT3'-UTR融合的荧光素酶活性较对照组降低,差异有统计学意义(P<0.05),而miR-124模拟转染组与Sp1-MUT的3'-UTR融合的荧光素酶活性与对照组相比,差异无统计学意义(P>0.05)。miR-124模拟转染组α-SMA、SM22a、calponin和MYH11蛋白表达较对照组升高,miR-124-pCMV-Sp1Mut共转染组α-SMA、SM22a、calponin和MYH11蛋白表达较miR-124模拟转染组降低,差异均有统计学意义(P<0.05)。miR-124模拟转染组Sp1蛋白表达较对照组降低,miR-124-pCMV-Sp1Mut共转染组Sp1蛋白表达较miR-124模拟转染组升高,差异均有统计学意义(P<0.05)。结论miR-124通过Sp1调节HVSMC的表型并显著抑制IA发展,miR-124对Sp1表达的调节可能为miR-124的功能脑血管疾病的分子机制提供新的见解。
