目的:研究miR-1271在卵巢癌SKOV3/DDP细胞顺铂耐药形成中的作用。方法:采用实时定量PCR法检测卵巢癌SKOV3/DDP细胞和母代SKOV3细胞中miR-1271的表达,并通过蛋白质印迹法检测上述细胞X连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis ...目的:研究miR-1271在卵巢癌SKOV3/DDP细胞顺铂耐药形成中的作用。方法:采用实时定量PCR法检测卵巢癌SKOV3/DDP细胞和母代SKOV3细胞中miR-1271的表达,并通过蛋白质印迹法检测上述细胞X连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)的表达。构建XIAP 3'UTR荧光素酶报告质粒验证miR-1271的靶基因,在耐药细胞株中瞬时转染miR-1271模拟物以观察上调miR-1271对卵巢癌SKOV3/DDP细胞XIAP表达及顺铂耐药的影响,并运用流式细胞术检测转染miR-1271模拟物后SKOV3/DDP细胞的凋亡率。结果:miR-1271在卵巢癌SKOV3/DDP细胞中呈低表达,抗凋亡蛋白XIAP在卵巢癌SKOV3/DDP细胞中呈高表达,荧光素酶报告实验证实XIAP是直接受miR-1271调控的靶基因,在耐药株中上调miR-1271显著抑制XIAP蛋白表达水平,显著增加细胞对顺铂的敏感性,并显著增加细胞对顺铂诱导的凋亡。结论:miR-1271靶向抑制XIAP表达可增加卵巢癌SKOV3/DDP细胞对顺铂的敏感性。展开更多
目的探讨miR-1271对卵巢癌细胞的增殖、侵袭和迁移的影响,及其作用机制。方法实验分为miR-NC组(转染miR-NC)、miR-1271组(转染miR-1271 mimics)、miR-1271+pcDNA3.1组(共转染miR-1271和pcDNA3.1)、miR-1271+pcDNA3.1-Twist1组(共转染miR...目的探讨miR-1271对卵巢癌细胞的增殖、侵袭和迁移的影响,及其作用机制。方法实验分为miR-NC组(转染miR-NC)、miR-1271组(转染miR-1271 mimics)、miR-1271+pcDNA3.1组(共转染miR-1271和pcDNA3.1)、miR-1271+pcDNA3.1-Twist1组(共转染miR-1271和pcDNA3.1-Twist1)。用噻唑蓝法检测细胞活性,用Transwell检测细胞迁移和侵袭情况,用实时定量聚合酶链反应检测miR-1271和碱性螺旋-环-螺旋转录因子1(Twist1)mRNA的表达水平,用荧光素酶报告基因检测实验检测miR-1271对Twist1的靶向调控。结果干预24 h后,miR-NC组、miR-1271组、miR-1271+pcDNA3.1组和miR-1271+pcDNA3.1-Twist1组的细胞活性分别为0.71±0.07,0.46±0.04,0.42±0.04和0.57±0.05,迁移细胞数分别为(129.00±9.69),(68.00±3.26),(67.00±3.18)和(98.00±6.85)个,侵袭细胞数分别为(110.00±8.16),(53.00±2.91),(55.00±2.96)和(83.00±4.39)个,miR-1271表达水平分别为0.18±0.02,0.47±0.04,0.46±0.06和0.49±0.01,Twist1 mRNA表达水平分别为0.68±0.06,0.13±0.01,0.11±0.01和0.47±0.03。miR-1271组和miR-1271+pcDNA3.1-Twist1组的细胞活性、迁移和侵袭细胞数、miR-1271和Twist1 mRNA表达水平分别与miR-NC组和miR-1271+pcDNA3.1组比较,差异均有统计学意义(均P<0.05)。转染WT-Twist1后,相较于miR-NC组,miR-1271组的荧光素酶活性显著降低(0.63±0.06 vs 1.12±0.10,P<0.05);而转染MUT-Twist1后,相较于miR-con组,miR-1271组的荧光素酶活性无显著变化(1.09±0.10 vs 1.11±0.10,P>0.05)。结论miR-1271可抑制卵巢癌细胞的增殖、迁移和侵袭,其机制可能与Twist1有关。展开更多
[目的]探讨miR-1271对甲状腺乳头状癌细胞增殖和转移的影响及其作用机制。[方法]收集20例手术切除甲状腺乳头状癌患者的癌组织和对应的癌旁组织标本。采用RTqPCR检测组织和细胞系中miR-1271和ZEB1的表达水平;CCK-8检测K1细胞增殖活力;Tr...[目的]探讨miR-1271对甲状腺乳头状癌细胞增殖和转移的影响及其作用机制。[方法]收集20例手术切除甲状腺乳头状癌患者的癌组织和对应的癌旁组织标本。采用RTqPCR检测组织和细胞系中miR-1271和ZEB1的表达水平;CCK-8检测K1细胞增殖活力;Transwell检测K1细胞侵袭和迁移能力;Western blot检测蛋白的表达水平;双荧光素酶报告基因验证miR-1271与ZEB1的靶向关系。[结果]miR-1271在甲状腺乳头状癌组织中的表达水平低于癌旁组织组织(1.175±0.534 vs 3.918±0.512,P<0.001)。与人甲状腺滤泡上皮正常细胞(0.985±0.062)相比,miR-1271在癌细胞系(TPC-1:0.752±0.052,K1:0.318±0.042,BCPAP:0.584±0.045)中的表达水平明显下调(P均<0.05),且在K1细胞中表达最低(P=0.016)。双荧光素酶报告基因结果显示,miR-1271通过结合ZEB1基因的3′UTR,进而抑制其表达水平。过表达ZEB1可明显缓解miR-1271对K1细胞增殖和转移的抑制作用。[结论]miR-1271通过靶向下调ZEB1的表达,进而抑制K1细胞增殖、侵袭、迁移和EMT。展开更多
BACKGROUND Hepatocellular carcinoma(HCC)is the third leading cause of cancer death,causing about 750000 deaths worldwide every year.Patients with advanced hepatocellular carcinoma will often only receive transcatheter...BACKGROUND Hepatocellular carcinoma(HCC)is the third leading cause of cancer death,causing about 750000 deaths worldwide every year.Patients with advanced hepatocellular carcinoma will often only receive transcatheter arterial chemoembolization(TACE).Glypican-3(GPC3)is one of the most promising serum markers for HCC.Abnormal expression of miRNAs may be involved in the occurrence and development of tumor.AIM To explore the value of miR-1271 and GPC3 in evaluating the prognosis of patients with HCC after TACE.METHODS From January 2016 to December 2018,162 patients with advanced HCC who received TACE in our hospital were selected into the cancer group,and 162 patients who underwent physical examination during the same period were selected into the health group.The patients in the HCC group were treated with TACE.The changes of serum GPC3 and circulating miR-1271 in the HCC before and after TACE were analyzed.The expression of serum GPC3 was detected by enzyme-linked immunosorbent assay,and the expression of circulating miR-1271 was detected by real-time quantitative polymerase chain reaction.The methodological results of sensitivity,specificity,and accuracy of miR-1271 and GPC3 alone and joint detection of HCC were also evaluated.RESULTSThe level of serum GPC3 in patients with HCC was significantly higher than that in healthy controls.GPC3 levels were increased in both HCC patients and those treated with TACE compared with healthy controls.After TACE,the level of serum GPC3 was significantly lower than that before treatment(P<0.05),and the level of circulating miR-1271 was significantly higher than that before treatment(P<0.05).There were 112 cases(69.14%)with remission(complete remission+complete remission+stable disease)and 50 cases(30.86%)with relapse disease progression in HCC patients.After TACE,the miR-1271 level in patients with remission and relapse was lower than that in the healthy group,and the GPC3 level was higher than that in the healthy group,the differences were statistically significant(P<0.05).The 展开更多
文摘目的探讨miR-1271对卵巢癌细胞的增殖、侵袭和迁移的影响,及其作用机制。方法实验分为miR-NC组(转染miR-NC)、miR-1271组(转染miR-1271 mimics)、miR-1271+pcDNA3.1组(共转染miR-1271和pcDNA3.1)、miR-1271+pcDNA3.1-Twist1组(共转染miR-1271和pcDNA3.1-Twist1)。用噻唑蓝法检测细胞活性,用Transwell检测细胞迁移和侵袭情况,用实时定量聚合酶链反应检测miR-1271和碱性螺旋-环-螺旋转录因子1(Twist1)mRNA的表达水平,用荧光素酶报告基因检测实验检测miR-1271对Twist1的靶向调控。结果干预24 h后,miR-NC组、miR-1271组、miR-1271+pcDNA3.1组和miR-1271+pcDNA3.1-Twist1组的细胞活性分别为0.71±0.07,0.46±0.04,0.42±0.04和0.57±0.05,迁移细胞数分别为(129.00±9.69),(68.00±3.26),(67.00±3.18)和(98.00±6.85)个,侵袭细胞数分别为(110.00±8.16),(53.00±2.91),(55.00±2.96)和(83.00±4.39)个,miR-1271表达水平分别为0.18±0.02,0.47±0.04,0.46±0.06和0.49±0.01,Twist1 mRNA表达水平分别为0.68±0.06,0.13±0.01,0.11±0.01和0.47±0.03。miR-1271组和miR-1271+pcDNA3.1-Twist1组的细胞活性、迁移和侵袭细胞数、miR-1271和Twist1 mRNA表达水平分别与miR-NC组和miR-1271+pcDNA3.1组比较,差异均有统计学意义(均P<0.05)。转染WT-Twist1后,相较于miR-NC组,miR-1271组的荧光素酶活性显著降低(0.63±0.06 vs 1.12±0.10,P<0.05);而转染MUT-Twist1后,相较于miR-con组,miR-1271组的荧光素酶活性无显著变化(1.09±0.10 vs 1.11±0.10,P>0.05)。结论miR-1271可抑制卵巢癌细胞的增殖、迁移和侵袭,其机制可能与Twist1有关。
文摘[目的]探讨miR-1271对甲状腺乳头状癌细胞增殖和转移的影响及其作用机制。[方法]收集20例手术切除甲状腺乳头状癌患者的癌组织和对应的癌旁组织标本。采用RTqPCR检测组织和细胞系中miR-1271和ZEB1的表达水平;CCK-8检测K1细胞增殖活力;Transwell检测K1细胞侵袭和迁移能力;Western blot检测蛋白的表达水平;双荧光素酶报告基因验证miR-1271与ZEB1的靶向关系。[结果]miR-1271在甲状腺乳头状癌组织中的表达水平低于癌旁组织组织(1.175±0.534 vs 3.918±0.512,P<0.001)。与人甲状腺滤泡上皮正常细胞(0.985±0.062)相比,miR-1271在癌细胞系(TPC-1:0.752±0.052,K1:0.318±0.042,BCPAP:0.584±0.045)中的表达水平明显下调(P均<0.05),且在K1细胞中表达最低(P=0.016)。双荧光素酶报告基因结果显示,miR-1271通过结合ZEB1基因的3′UTR,进而抑制其表达水平。过表达ZEB1可明显缓解miR-1271对K1细胞增殖和转移的抑制作用。[结论]miR-1271通过靶向下调ZEB1的表达,进而抑制K1细胞增殖、侵袭、迁移和EMT。
文摘BACKGROUND Hepatocellular carcinoma(HCC)is the third leading cause of cancer death,causing about 750000 deaths worldwide every year.Patients with advanced hepatocellular carcinoma will often only receive transcatheter arterial chemoembolization(TACE).Glypican-3(GPC3)is one of the most promising serum markers for HCC.Abnormal expression of miRNAs may be involved in the occurrence and development of tumor.AIM To explore the value of miR-1271 and GPC3 in evaluating the prognosis of patients with HCC after TACE.METHODS From January 2016 to December 2018,162 patients with advanced HCC who received TACE in our hospital were selected into the cancer group,and 162 patients who underwent physical examination during the same period were selected into the health group.The patients in the HCC group were treated with TACE.The changes of serum GPC3 and circulating miR-1271 in the HCC before and after TACE were analyzed.The expression of serum GPC3 was detected by enzyme-linked immunosorbent assay,and the expression of circulating miR-1271 was detected by real-time quantitative polymerase chain reaction.The methodological results of sensitivity,specificity,and accuracy of miR-1271 and GPC3 alone and joint detection of HCC were also evaluated.RESULTSThe level of serum GPC3 in patients with HCC was significantly higher than that in healthy controls.GPC3 levels were increased in both HCC patients and those treated with TACE compared with healthy controls.After TACE,the level of serum GPC3 was significantly lower than that before treatment(P<0.05),and the level of circulating miR-1271 was significantly higher than that before treatment(P<0.05).There were 112 cases(69.14%)with remission(complete remission+complete remission+stable disease)and 50 cases(30.86%)with relapse disease progression in HCC patients.After TACE,the miR-1271 level in patients with remission and relapse was lower than that in the healthy group,and the GPC3 level was higher than that in the healthy group,the differences were statistically significant(P<0.05).The
