食管癌组织miR-16-5p和FASN表达水平及临床意义

Study on the expression levels and clinical significance of miR-16-5p and FASN in esophageal carcinoma

目的探索微小RNA-16-5p(miR-16-5p)、脂肪酸合成酶(FASN)在食管癌组织中的表达水平及临床意义。方法选取2017年2月-2020年1月上海健康医学院附属崇明医院收治的104例食管癌患者为研究对象,术中收集其癌组织与癌旁组织并采用实时荧光定量PCR(qRT-PCR)测定组织miR-16-5p、FASN表达量;免疫组化法检测组织中FASN蛋白表达情况;TargetScanHuman网站预测miR-16-5p与FASN的靶向关系;Pearson法分析食管癌患者癌组织miR-16-5p与FASN水平的相关性;采用双荧光素酶报告实验验证miR-16-5p与FASN的靶向关系。依据miR-16-5p、FASN表达量平均值将食管癌患者分为miR-16-5p高表达组(≥0.82,n=53)与miR-16-5p低表达组(<0.82,n=51),FASN高表达组(≥1.23,n=55)与FASN低表达组(<1.23,n=49),采用Kaplan-Meier生存曲线分析食管癌组织miR-16-5p、FASN表达情况与患者生存率之间的关系;采用Cox回归分析食管癌患者预后不良的危险因素。结果食管癌组织中miR-16-5p表达水平(0.82±0.14)低于癌旁组织(0.98±0.11),FASN表达(1.23±0.10)高于癌旁组织(1.01±0.09),差异均有统计学意义(t=9.164、16.676,P均<0.001);FASN蛋白在食管癌组织中阳性表达率为61.54%(64/104),明显高于癌旁组织的17.31%(18/104),差异有统计学意义(χ^(2)=42.599,P<0.05)。TargetScanHuman网站预测miR-16-5p与FASN存在结合位点;食管癌患者癌组织中miR-16-5p与FASN表达水平成负相关(r=-0.584,P<0.01),双荧光素酶报告实验证实miR-16-5p与FASN存在靶向关系(F=144.623,P<0.001)。食管癌组织miR-16-5p、FASN表达均与肿瘤直径(χ^(2)=5.525、3.905,P=0.019、0.048)、浸润深度(χ^(2)=8.255、7.638,P=0.004、0.006)、TNM分期(χ^(2)=8.792、8.521,P=0.003、0.004)、淋巴结转移(χ^(2)=8.054、9.697,P=0.005、0.002)、组织分化程度(χ^(2)=4.695、4.579,P=0.030、0.032)有关。miR-16-5p高表达组食管癌患者3年生存率(73.58%)高于miR-16-5p低表达组(43.14%),FASN高表达组食管癌患者3年生存率(45.45%)低于FASN低表达组(73.47%),差异均有统计学意义(χ^(2)=9.935、8.386,P=0.002、0.004)。多因素Cox回归分析表示,深层浸润(HR=2.116,P=0.033)、TNM分期为Ⅲ/Ⅳ期(HR=2.885,P=0.032)、淋巴结转移(HR=3.207,P=0.027)、FASN高表达(HR=4.104,P=0.006)均是食管癌患者预后不良的危险因素,miR-16-5p高表达是食管癌预后的保护因素(HR=0.559,P=0.012)。结论食管癌组织中miR-16-5p表达水平下调、FASN表达水平上调,二者均与食管癌患者临床病理特征及预后紧密相关。

Objective To explore the expression level and clinical significance of microRNA-16-5p(miR-16-5p)and fatty acid synthase(FASN)in esophageal cancer.Methods A total of 104 patients with esophageal cancer admitted to the Shanghai Health Medical College Affiliated Chongming Hospital from February 2017 to January 2020 were regarded as the study subjects,the expression of miR-16-5p and FASN was measured by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)technique after surgical collection of tumor and adjacent tissues;immunohistochemical method was applied to detect the expression of FASN protein in tissues;TargetScanHuman website was applied to predict the targeting relationship between miR-16-5p and FASN;Pearson method was applied to analyze the correlation between miR-16-5p and FASN levels in esophageal cancer tissues.The targeting relationship between miR-16-5p and FASN was verified by double luciferase reporting experiment.According to the average expression of miR-16-5p and FASN,the patients with esophageal carcinoma were divided into high expression of miR-16-5p group(≥0.82,n=53)and low expression of miR-16-5p group(<0.82,n=51),high expression of FASN group(≥1.23,n=55)and low expression of FASN group(<1.23,n=49).Kaplan-Meier survival curve was applied to analyze the relationship between the expression of miR-16-5p and FASN in esophageal cancer tissue and the survival rate of patients;Cox regression was applied to analyze the risk factors for poor prognosis in patients with esophageal cancer.Results The expression level of miR-16-5p in esophageal cancer(0.82±0.14)was lower than that in adjacent tissues(0.98±0.11),while the expression of FASN(1.23±0.10)was higher than that in adjacent tissues(1.01±0.09),and the differences were statistically significant(t=9.164,16.676,all P<0.001).The positive expression rate of FASN protein in esophageal carcinoma was 61.54%(64/104),which was significantly higher than that in adjacent tissues[17.31%(18/104)],the difference was statistically significant(χ^(2)=42.599,P<0.05).The expression of miR-16-5p had binding site with FASN by TargetScanHuman website prediction;miR-16-5p was negatively correlated with FASN expression level in cancer tissues of esophageal carcinoma patients(r=-0.584,P<0.01),the double Luciferase report experiment confirmed that there was a targeting relationship between miR-16-5p and FASN(F=144.623,P<0.001).The expression of miR-16-5p and FASN in esophageal carcinoma was correlated with tumor diameter(χ^(2)=5.525,3.905;P=0.019,0.048),depth of invasion(χ^(2)=8.255,7.638;P=0.004,0.006),TNM stage(χ^(2)=8.792,8.521;P=0.003,0.004),lymph node metastasis(χ^(2)=8.054,9.697;P=0.005,0.002),and degree of tissue differentiation(χ^(2)=4.695,4.579;P=0.030,0.032).The 3-year survival rate of patients in high expression of miR-16-5p group in esophageal cancer(73.58%)was higher than that of low expression of miR-16-5p group(43.14%);the 3-year survival rate of esophageal cancer patients in high expression of FASN group(45.45%)was lower than that of low expression of FASN group(73.47%);the differences were statistically significant(χ^(2)=9.935,8.386;P=0.002,0.004).Multivariate Cox regression analysis showed that deep invasion(HR=2.116,P=0.033),TNM stageⅢ/Ⅳ(HR=2.885,P=0.032),lymph node metastasis(HR=3.207,P=0.027),high expression of FASN(HR=4.104,P=0.006)were risk factors for poor prognosis in esophageal cancer patients;high expression of miR-16-5p(HR=0.559,P=0.012)was a protective factor.Conclusion The expression level of miR-16-5p was down regulated and the expression level of FASN was up regulated in esophageal cancer tissue;both of which were closely related to the clinicopathological characteristics and prognosis of esophageal cancer patients.