目的了解MicroRNA-153(miR-153)在膀胱癌组织中的表达及对膀胱癌细胞生物学特性的影响。方法收集经病理确诊为膀胱癌进行外科手术切除的25例患者的膀胱癌组织及距离膀胱癌病灶5 cm以上的癌旁组织,通过实时定量聚合酶链反应(Real time P...目的了解MicroRNA-153(miR-153)在膀胱癌组织中的表达及对膀胱癌细胞生物学特性的影响。方法收集经病理确诊为膀胱癌进行外科手术切除的25例患者的膀胱癌组织及距离膀胱癌病灶5 cm以上的癌旁组织,通过实时定量聚合酶链反应(Real time PCR)检测miR-153在25对膀胱癌及瘤旁组织中的表达,分析miR-153基因表达与肿瘤分类(肌层浸润型与非肌层浸润型)及病理分级的关系。采用Annexin V/PI双染法和MTT实验绘制细胞生长曲线,采用生物信息学软件及荧光素酶实验验证miR-153是否存在直接作用。结果与癌旁组织相比,miR-153在膀胱癌组织中的相对表达量分别为1.04±0.07和0.14±0.03。膀胱癌组织显著低于癌旁组织,差异有统计学意义(P<0.05);miR-153在肌层浸润组中的表达为非肌层浸润组的(2.48±0.32)倍,其表达水平随着病理分级升高而升高(P<0.05);miR-153成熟体的膀胱癌EJ细胞中RAS蛋白的表达水平较对照组显著降低,且具有浓度依赖性。结论miR-153在膀胱癌组织中的相对表达量明显升高,且其表达水平随着病理分级升高而升高。由于受到脂质体毒性的影响,肿瘤细胞的增殖受到抑制,anti-miRl53组的活性细胞数明显下降。anti-miRl53特异性地抑制了膀胱癌细胞的增殖。特异性anti-miRl53的转染使膀胱癌细胞发生了包括凋亡和坏死在内的肿瘤细胞的死亡,提示有可能参与了膀胱癌细胞的增殖和凋亡过程,并可作为膀胱癌的潜在临床诊断标志物。展开更多
BACKGROUND Exosomes contain proteins, lipids, and biological molecules such as DNA and RNA. Nucleic acids in exosomes are a group of molecules that can act as biomarkers. Currently, there are many reports on exosomal ...BACKGROUND Exosomes contain proteins, lipids, and biological molecules such as DNA and RNA. Nucleic acids in exosomes are a group of molecules that can act as biomarkers. Currently, there are many reports on exosomal microRNAs, which are ideal biomarkers for the early diagnosis of cancer. However, there are few reports on the role of exosomal microRNAs in the diagnosis and prognosis of hepatocellular carcinoma(HCC).AIM To understand the mechanism of exosomal microRNA-224(miR-224) in the development of HCC and evaluate its diagnostic and prognostic value.METHODS Cell culture and transfection of exosomal miRNA-224, real-time quantitative PCR, luciferase reporter assay, and other methods were used to find new biomarkers related to the development of HCC that can be used to diagnose HCC and predict HCC prognosis.RESULTSBy targeting glycine N-methyltransferase, incubating exosomes with miR-224 mimic resulted in a significant increase in cell proliferation compared to that of the control group, while incubation with the miR-224 inhibitor significantly reduced cell proliferation. The same results were obtained for the cell invasion assay. Serum exosomal miR-224 did have some ability to differentiate patients with HCC from healthy controls, with an area under the curve of 0.910, and HCC patients with higher serum exosomal miR-224 expression had lower overall survival.CONCLUSION Exosomal miR-224 is a tumor promotor and can be a marker of diagnosis and prognosis of HCC patients, however, its ability to distinguish liver diseases needs further verification.展开更多
Background:Crohn’s disease(CD)has a tendency for recurrence and requires adequate monitoring and personalized treatment.Since endoscopy is considerably invasive,serum biomarkers are required as alternatives for CD mo...Background:Crohn’s disease(CD)has a tendency for recurrence and requires adequate monitoring and personalized treatment.Since endoscopy is considerably invasive,serum biomarkers are required as alternatives for CD monitoring.Toward this,exosomal microRNAs(miRNAs)may serve as promising candidates.In this study,we aimed to assess the role of serum exosomal microRNA-144-3p(miR-144-3p)as a biomarker for CD monitoring.Methods:We prospectively recruited 154 patients without a history of surgery(Cohort 1)and 75 patients who were to undergo intestinal resection(Cohort 2).Serum samples were collected from Cohort 1 before colonoscopy and from Cohort 2 before surgery and during post-operative colonoscopic examination.The serum levels of exosomal miR-144-3p were measured using quantitative reverse-transcription polymerase chain reaction(PCR).Correlations between relative exosomal miR-144-3p levels,disease activity,and disease behavior were analysed.The area under the receiver-operating characteristic curve(AUC)was used to assess the predictive value of exosomal miR-144-3p regarding mucosal activity and postoperative recurrence.Results:A 3.33-fold increase in serum exosomal miR-144-3p levels was recorded in patients with CD compared with those in healthy controls(P<0.001).The exosomalmiR-144-3p levels were positively correlated with the simple endoscopic score of CD(q=0.547,P<0.001)as well as the Rutgeerts score(q=0.478,P<0.001).Elevated exosomalmiR-144-3p levels were correlated with the penetrating disease with high specificity(100%[95%confidence interval,95.1%–100%]).The accuracy of exosomalmiR-144-3p for identifying post-operative recurrence was higher than that of C-reactive protein(CRP)(AUC,0.775 vs 0.639;P<0.001).Conclusions:Serum exosomal miR-144-3p is a reliable biomarker of mucosal inflammation and penetrating CD.It may identify endoscopic CD recurrence after intestinal resection with higher accuracy than CRP testing.展开更多
文摘目的了解MicroRNA-153(miR-153)在膀胱癌组织中的表达及对膀胱癌细胞生物学特性的影响。方法收集经病理确诊为膀胱癌进行外科手术切除的25例患者的膀胱癌组织及距离膀胱癌病灶5 cm以上的癌旁组织,通过实时定量聚合酶链反应(Real time PCR)检测miR-153在25对膀胱癌及瘤旁组织中的表达,分析miR-153基因表达与肿瘤分类(肌层浸润型与非肌层浸润型)及病理分级的关系。采用Annexin V/PI双染法和MTT实验绘制细胞生长曲线,采用生物信息学软件及荧光素酶实验验证miR-153是否存在直接作用。结果与癌旁组织相比,miR-153在膀胱癌组织中的相对表达量分别为1.04±0.07和0.14±0.03。膀胱癌组织显著低于癌旁组织,差异有统计学意义(P<0.05);miR-153在肌层浸润组中的表达为非肌层浸润组的(2.48±0.32)倍,其表达水平随着病理分级升高而升高(P<0.05);miR-153成熟体的膀胱癌EJ细胞中RAS蛋白的表达水平较对照组显著降低,且具有浓度依赖性。结论miR-153在膀胱癌组织中的相对表达量明显升高,且其表达水平随着病理分级升高而升高。由于受到脂质体毒性的影响,肿瘤细胞的增殖受到抑制,anti-miRl53组的活性细胞数明显下降。anti-miRl53特异性地抑制了膀胱癌细胞的增殖。特异性anti-miRl53的转染使膀胱癌细胞发生了包括凋亡和坏死在内的肿瘤细胞的死亡,提示有可能参与了膀胱癌细胞的增殖和凋亡过程,并可作为膀胱癌的潜在临床诊断标志物。
基金Supported by the Scientific and Technological Research Project of Henan Province,No.162102310024
文摘BACKGROUND Exosomes contain proteins, lipids, and biological molecules such as DNA and RNA. Nucleic acids in exosomes are a group of molecules that can act as biomarkers. Currently, there are many reports on exosomal microRNAs, which are ideal biomarkers for the early diagnosis of cancer. However, there are few reports on the role of exosomal microRNAs in the diagnosis and prognosis of hepatocellular carcinoma(HCC).AIM To understand the mechanism of exosomal microRNA-224(miR-224) in the development of HCC and evaluate its diagnostic and prognostic value.METHODS Cell culture and transfection of exosomal miRNA-224, real-time quantitative PCR, luciferase reporter assay, and other methods were used to find new biomarkers related to the development of HCC that can be used to diagnose HCC and predict HCC prognosis.RESULTSBy targeting glycine N-methyltransferase, incubating exosomes with miR-224 mimic resulted in a significant increase in cell proliferation compared to that of the control group, while incubation with the miR-224 inhibitor significantly reduced cell proliferation. The same results were obtained for the cell invasion assay. Serum exosomal miR-224 did have some ability to differentiate patients with HCC from healthy controls, with an area under the curve of 0.910, and HCC patients with higher serum exosomal miR-224 expression had lower overall survival.CONCLUSION Exosomal miR-224 is a tumor promotor and can be a marker of diagnosis and prognosis of HCC patients, however, its ability to distinguish liver diseases needs further verification.
基金supported by the National Natural Science Foundation of China[grant numbers 81630018,82070538,,81870374]Guangdong Science and Technology Department[grant number 2017A030306021]Guangzhou Science and Technology Department[grant number 202002030041].
文摘Background:Crohn’s disease(CD)has a tendency for recurrence and requires adequate monitoring and personalized treatment.Since endoscopy is considerably invasive,serum biomarkers are required as alternatives for CD monitoring.Toward this,exosomal microRNAs(miRNAs)may serve as promising candidates.In this study,we aimed to assess the role of serum exosomal microRNA-144-3p(miR-144-3p)as a biomarker for CD monitoring.Methods:We prospectively recruited 154 patients without a history of surgery(Cohort 1)and 75 patients who were to undergo intestinal resection(Cohort 2).Serum samples were collected from Cohort 1 before colonoscopy and from Cohort 2 before surgery and during post-operative colonoscopic examination.The serum levels of exosomal miR-144-3p were measured using quantitative reverse-transcription polymerase chain reaction(PCR).Correlations between relative exosomal miR-144-3p levels,disease activity,and disease behavior were analysed.The area under the receiver-operating characteristic curve(AUC)was used to assess the predictive value of exosomal miR-144-3p regarding mucosal activity and postoperative recurrence.Results:A 3.33-fold increase in serum exosomal miR-144-3p levels was recorded in patients with CD compared with those in healthy controls(P<0.001).The exosomalmiR-144-3p levels were positively correlated with the simple endoscopic score of CD(q=0.547,P<0.001)as well as the Rutgeerts score(q=0.478,P<0.001).Elevated exosomalmiR-144-3p levels were correlated with the penetrating disease with high specificity(100%[95%confidence interval,95.1%–100%]).The accuracy of exosomalmiR-144-3p for identifying post-operative recurrence was higher than that of C-reactive protein(CRP)(AUC,0.775 vs 0.639;P<0.001).Conclusions:Serum exosomal miR-144-3p is a reliable biomarker of mucosal inflammation and penetrating CD.It may identify endoscopic CD recurrence after intestinal resection with higher accuracy than CRP testing.
