AIM To explore the effect of miR-382 on esophageal squamous cell carcinoma (ESCC) in vitro and its possible molecular mechanism. METHODS Eca 109 cells derived from human ESCC and Het-1A cells derived from human normal...AIM To explore the effect of miR-382 on esophageal squamous cell carcinoma (ESCC) in vitro and its possible molecular mechanism. METHODS Eca 109 cells derived from human ESCC and Het-1A cells derived from human normal esophageal epithelium were used. Lentivirus-mediated miR-382 was overexpressed in Eca109 cells. The effect of miR-382 on cell proliferation was evaluated by MTT and colony formation assay. For cell cycle analysis, cells were fixed and stained for 30 min with propidium iodide (PI) staining buffer containing 10 mg/mL PI and 100 mg/mL RNase A, and analyzed by BD FACSCalibur (TM) flow cytometer. For cell apoptosis assay, cells were stained with an Annexin V-FITC/PI Apoptosis Detection Kit according to the manufacturer's instructions and analyzed by a dual-laser flow cytometer. Cell invasion and migration abilities were determined through use of transwell chambers, non-coated or pre-coated with matrigel. Levels of proteins related to cell growth and migration were examined by western blotting. RESULTS Endogenous miR-382 was down-regulated in Eca109 cells compared with Het-1A. Introduction of miR-382 not only significantly inhibited proliferation and colony formation, but also arrested cell cycle at the G2/M phase, as well as promoted apoptosis and autophagy in Eca109 cells. Migration, invasion and epithelial-mesenchymal transition of Eca109 cells were suppressed by overexpressing miR-382. Western blotting results showed that miR-382 inhibited the phosphorylation of mTOR and 4E-BP1. CONCLUSION miR-382 functions as a tumor suppressor against ESCC development and metastasis, and could be considered as a potential drug source for the treatment of ESCC patients.展开更多
AIM: To study the potential prognostic role of microRNA-382(miR-382) in esophageal squamous cell carcinoma(ESCC).METHODS: Forty six patients were divided into 2groups according to postoperative survival time:the poor ...AIM: To study the potential prognostic role of microRNA-382(miR-382) in esophageal squamous cell carcinoma(ESCC).METHODS: Forty six patients were divided into 2groups according to postoperative survival time:the poor outcome group(28 patients), who showed early metastasis but no recurrence, and died within 1year after surgery, 12 patients of the group received postoperative chemotherapy treatment that was given after early metastasis happening; the good outcome group(18 patients), who had no clinical metastasis and recurrence, and survived 5 years or more after surgery, all patients did not receive any postoperative treatment. Total RNA was extracted from the patients' formalin-fixed and paraffin-embedded esophageal cancer tissues. miR-382 level was evaluated using highthroughput real-time quantitative polymerase chain reaction analysis. The correlation between miR-382 level and clinicopathologic features was analyzed through COX regression model, and Kaplan-Meier analysis was used to analyze the relationship betweenmiR-382 level and patient survival time.RESULTS: miR-382 was differentially expressed in the two groups. Overall the average miR-382 level in the ESCC patients with good outcome was 9.8 ± 3.8,while miR-382 level in the ESCC patients with poor outcome was 3.0 ± 0.8. The differences of miR-382 levels between two groups were significant(P < 0.05).Kaplan-Meier analysis results showed that miR-382 expression level generally had a significant reversecorrelation with ESCC patient survival time(P <0.001), in which the patients with higher expressions of miR-382 had a longer survival time either among individuals with the same tumor stage or among the overall patients.CONCLUSION: miR-382 levels are reverse-correlated with ESCC poor outcomes, suggesting that miR-382 could be a potential predictive biomarker for both prognosis and treatment of ESCC.展开更多
目的探讨血清miR-382-5p在丙型肝炎病毒(hepatitis C virus,HCV)相关肝细胞癌(hepatocellular carcinoma,HCC)患者中的表达及其联合甲胎蛋白(alpha fetoprotein,AFP)、异常凝血酶原-Ⅱ(protein induced by vitamin K antagonist-Ⅱ,PIV...目的探讨血清miR-382-5p在丙型肝炎病毒(hepatitis C virus,HCV)相关肝细胞癌(hepatocellular carcinoma,HCC)患者中的表达及其联合甲胎蛋白(alpha fetoprotein,AFP)、异常凝血酶原-Ⅱ(protein induced by vitamin K antagonist-Ⅱ,PIVKA-Ⅱ)对HCC诊断的价值。方法选取2019年1月至2022年3月保定市人民医院收治的102例HCV相关HCC患者(HCC组)、55例肝硬化患者(肝硬化组)和45例体检正常者(对照组)为研究对象,比较各组血清miR-382-5p、AFP及PIVKA-Ⅱ水平,比较不同临床特征HCC患者血清miR-382-5p的表达水平。应用受试者工作特征(receiver operator characteristic,ROC)曲线分析血清miR-382-5p、AFP及PIVKA-Ⅱ水平对HCC的诊断价值。采用Pearson相关分析HCC患者血清miR-382-5p表达水平与AFP及PIVKA-Ⅱ的相关性。结果HCC组血清miR-382-5p(6.83±2.51比3.15±1.08比1.82±0.76)、AFP[(527.10±73.26)μg/L比(22.84±9.25)μg/L比(3.17±0.36)μg/L]及PIVKA-Ⅱ[(352.64±69.12)mAU/ml比(21.73±6.90)mAU/ml比(18.30±4.27)mAU/ml]水平均显著高于肝硬化组和对照组(P均<0.05)。TNM分期Ⅲ~Ⅳ期(8.70±3.51比5.06±1.80)、Child-Pugh分级C级(8.14±2.90比5.62±1.91)、低分化(7.68±2.70比6.11±2.08)、门静脉癌栓(7.74±2.73比6.04±2.01)及远处转移(8.06±2.84比5.71±1.95)的HCC患者血清miR-382-5p表达水平显著升高(P均<0.05)。miR-382-5p、AFP及PIVKA-Ⅱ三项联合诊断HCC的ROC曲线下面积最大(0.980,95%CI:0.918~0.997),其敏感度最高(98.5%)。相关分析显示,HCC患者血清miR-382-5p水平与AFP(r=0.795,P<0.001)和PIVKA-Ⅱ(r=0.866,P<0.001)均呈正相关。结论血清miR-382-5p水平在HCV相关HCC患者中显著升高,联合AFP及PIVKA-Ⅱ检测可提高HCC的诊断价值。展开更多
基金Supported by Key Technologies R&D Program of Science and Technology Commission of Henan Province,No.152102310110 to Zhao BSKey Science and Technique Fund of Xinxiang,No.ZG15018 to Zhao BS
文摘AIM To explore the effect of miR-382 on esophageal squamous cell carcinoma (ESCC) in vitro and its possible molecular mechanism. METHODS Eca 109 cells derived from human ESCC and Het-1A cells derived from human normal esophageal epithelium were used. Lentivirus-mediated miR-382 was overexpressed in Eca109 cells. The effect of miR-382 on cell proliferation was evaluated by MTT and colony formation assay. For cell cycle analysis, cells were fixed and stained for 30 min with propidium iodide (PI) staining buffer containing 10 mg/mL PI and 100 mg/mL RNase A, and analyzed by BD FACSCalibur (TM) flow cytometer. For cell apoptosis assay, cells were stained with an Annexin V-FITC/PI Apoptosis Detection Kit according to the manufacturer's instructions and analyzed by a dual-laser flow cytometer. Cell invasion and migration abilities were determined through use of transwell chambers, non-coated or pre-coated with matrigel. Levels of proteins related to cell growth and migration were examined by western blotting. RESULTS Endogenous miR-382 was down-regulated in Eca109 cells compared with Het-1A. Introduction of miR-382 not only significantly inhibited proliferation and colony formation, but also arrested cell cycle at the G2/M phase, as well as promoted apoptosis and autophagy in Eca109 cells. Migration, invasion and epithelial-mesenchymal transition of Eca109 cells were suppressed by overexpressing miR-382. Western blotting results showed that miR-382 inhibited the phosphorylation of mTOR and 4E-BP1. CONCLUSION miR-382 functions as a tumor suppressor against ESCC development and metastasis, and could be considered as a potential drug source for the treatment of ESCC patients.
基金Supported by Xinxiang Medical University Key Areas grant,No.ZD2011-8(to BS Zhao)
文摘AIM: To study the potential prognostic role of microRNA-382(miR-382) in esophageal squamous cell carcinoma(ESCC).METHODS: Forty six patients were divided into 2groups according to postoperative survival time:the poor outcome group(28 patients), who showed early metastasis but no recurrence, and died within 1year after surgery, 12 patients of the group received postoperative chemotherapy treatment that was given after early metastasis happening; the good outcome group(18 patients), who had no clinical metastasis and recurrence, and survived 5 years or more after surgery, all patients did not receive any postoperative treatment. Total RNA was extracted from the patients' formalin-fixed and paraffin-embedded esophageal cancer tissues. miR-382 level was evaluated using highthroughput real-time quantitative polymerase chain reaction analysis. The correlation between miR-382 level and clinicopathologic features was analyzed through COX regression model, and Kaplan-Meier analysis was used to analyze the relationship betweenmiR-382 level and patient survival time.RESULTS: miR-382 was differentially expressed in the two groups. Overall the average miR-382 level in the ESCC patients with good outcome was 9.8 ± 3.8,while miR-382 level in the ESCC patients with poor outcome was 3.0 ± 0.8. The differences of miR-382 levels between two groups were significant(P < 0.05).Kaplan-Meier analysis results showed that miR-382 expression level generally had a significant reversecorrelation with ESCC patient survival time(P <0.001), in which the patients with higher expressions of miR-382 had a longer survival time either among individuals with the same tumor stage or among the overall patients.CONCLUSION: miR-382 levels are reverse-correlated with ESCC poor outcomes, suggesting that miR-382 could be a potential predictive biomarker for both prognosis and treatment of ESCC.
文摘目的探讨血清miR-382-5p在丙型肝炎病毒(hepatitis C virus,HCV)相关肝细胞癌(hepatocellular carcinoma,HCC)患者中的表达及其联合甲胎蛋白(alpha fetoprotein,AFP)、异常凝血酶原-Ⅱ(protein induced by vitamin K antagonist-Ⅱ,PIVKA-Ⅱ)对HCC诊断的价值。方法选取2019年1月至2022年3月保定市人民医院收治的102例HCV相关HCC患者(HCC组)、55例肝硬化患者(肝硬化组)和45例体检正常者(对照组)为研究对象,比较各组血清miR-382-5p、AFP及PIVKA-Ⅱ水平,比较不同临床特征HCC患者血清miR-382-5p的表达水平。应用受试者工作特征(receiver operator characteristic,ROC)曲线分析血清miR-382-5p、AFP及PIVKA-Ⅱ水平对HCC的诊断价值。采用Pearson相关分析HCC患者血清miR-382-5p表达水平与AFP及PIVKA-Ⅱ的相关性。结果HCC组血清miR-382-5p(6.83±2.51比3.15±1.08比1.82±0.76)、AFP[(527.10±73.26)μg/L比(22.84±9.25)μg/L比(3.17±0.36)μg/L]及PIVKA-Ⅱ[(352.64±69.12)mAU/ml比(21.73±6.90)mAU/ml比(18.30±4.27)mAU/ml]水平均显著高于肝硬化组和对照组(P均<0.05)。TNM分期Ⅲ~Ⅳ期(8.70±3.51比5.06±1.80)、Child-Pugh分级C级(8.14±2.90比5.62±1.91)、低分化(7.68±2.70比6.11±2.08)、门静脉癌栓(7.74±2.73比6.04±2.01)及远处转移(8.06±2.84比5.71±1.95)的HCC患者血清miR-382-5p表达水平显著升高(P均<0.05)。miR-382-5p、AFP及PIVKA-Ⅱ三项联合诊断HCC的ROC曲线下面积最大(0.980,95%CI:0.918~0.997),其敏感度最高(98.5%)。相关分析显示,HCC患者血清miR-382-5p水平与AFP(r=0.795,P<0.001)和PIVKA-Ⅱ(r=0.866,P<0.001)均呈正相关。结论血清miR-382-5p水平在HCV相关HCC患者中显著升高,联合AFP及PIVKA-Ⅱ检测可提高HCC的诊断价值。
