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骨肉瘤是临床上常见的原发性恶性骨肿瘤,多发于儿童和青少年,年发病率为百万分之三[1]。骨肉瘤病人的5年生存率为60%~70%,当发生远处转移时可降至20%~30%[2]。尽管在过去几十年里,骨肉瘤的治疗取得了很大的进展,如广泛肿瘤切除术、新辅助放化疗等,但是病人5年生存率和无病生存率仍然较低[3]。因此寻找新的诊断及预后标志物势在必行。微小RNA(miRNAs)是一类高度保守的非编码RNA,其长度为19~25个核苷酸。miRNAs可与靶mRNA的3′-UTR互补结合,抑制基因转录。miRNAs在人类多种肿瘤中有异常表达,并参与细胞增殖、分化、迁移和凋亡等生理学过程,发挥促癌或抑癌作用[4]。miR-1225-5p在骨肉瘤组织和细胞系中的表达明显下调。miR-1225-5p可通过靶向SOX9基因抑制骨肉瘤细胞的增殖和侵袭,提示miR-1225-5p在骨肉瘤中起到抑癌作用[5]。既往研究发现,miR-1225-5p在甲状腺癌[6]、胰腺癌[7]和喉癌[8]等癌症中同样起到抑癌作用。血清miR-1225-5p对骨肉瘤病人诊断及预后的评估有何意义,既往少有报道。本研究采用RT-PCR检测了104例骨肉瘤病人血清中miR-1225-5p的表达水平,旨在探讨血清miR-1225-5p在骨肉瘤诊断及预后中的价值。
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miR-1225-5p的扩增曲线平滑,基线期、对数期、平台期明显,扩增效果满意(见图 1A)。溶解曲线呈单一峰,溶解温度在80~90 ℃之间,说明无引物二聚体或者其他非特异性扩增片段的产生,实验检测结果的特异度比较高(见图 1B)。骨肉瘤组、骨膜炎组和对照组的血清miR-1225-5p相对表达量分别为(0.42±0.08)(0.58±0.11)(0.62±0.09),组间比较差异有统计学意义(F=102.74,P < 0.01),两两比较发现,骨肉瘤组血清miR-1225-5p相对表达量均低于骨膜炎组和对照组(P < 0.01),骨膜炎组低于对照组(P < 0.05)。术后3个月,骨肉瘤病人血清miR-1225-5p相对表达量从术前的(0.42±0.08)升高至(0.59±0.10),差异有统计学意义(t=13.54,P < 0.01)。骨肉瘤病人血清miR-1225-5p表达水平与Enneking分期、肿瘤转移有关(P < 0.01和P < 0.05),而与性别、年龄、肿瘤部位、肿瘤直径和WHO骨肉瘤分类无关(P>0.05)(见表 1)。
临床特征 高表达组
(n=52)低表达组
(n=52)χ2 P 年龄/岁 ≥18
< 1829
2326
260.35 >0.05 性别 男
女28
2430
220.16 >0.05 肿瘤部位 四肢
躯干33
1929
230.64 >0.05 Enneking分期 Ⅱ
Ⅲ27
2513
397.96 < 0.01 WHO分类 普通型骨肉瘤 27 22 6.05 >0.05 血管扩张型骨肉瘤 4 7 小细胞型骨肉瘤 2 6 骨膜骨肉瘤 1 3 骨旁骨肉瘤 8 9 髓内高分化骨肉瘤 10 5 肿瘤直径/cm < 5
≥527
2521
311.39 >0.05 远处转移 是
否19
3332
206.50 < 0.05 表 1 骨肉瘤病人血清miR-1225-5p表达水平与临床特征的关系
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ROC曲线分析结果显示,血清miR-1225-5p诊断骨肉瘤的曲线下面积为0.785(95%CI:0.625~0.934)。当血清miR-1225-5p相对表达量为0.46时约登指数最大,此时灵敏度和特异度分别为89.35%和71.30%(见图 2)。
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用Kaplan-Meier分析和Log-Rank检验分析骨肉瘤病人的生存情况,结果显示高表达组OS和DFS均高于低表达组(P < 0.05和P < 0.01)(见图 3、4)。COX多因素分析结果显示,血清miR-1225-5p是影响骨肉瘤病人OS和DFS的独立因素(P < 0.05和P < 0.01)(见表 2)。
因素 OS DFS HR 95%CI P HR 95%CI P Enneking分期 3.645 1.695~6.301 < 0.05 4.165 1.569~6.301 < 0.01 肿瘤转移 2.265 1.253~4.937 < 0.05 2.795 1.396~5.236 < 0.05 miR-1225-5p 3.555 1.269~5.001 < 0.05 3.934 1.568~6.152 < 0.01 表 2 COX比例风险回归模型分析影响骨肉瘤病人预后的因素
血清miR-1225-5p在骨肉瘤病人诊断及预后中的价值
Value of serum miR-1225-5p in the diagnosis and prognosis of osteosarcoma
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摘要:
目的探讨血清miR-1225-5p在骨肉瘤病人诊断及预后中的价值。 方法选取104例骨肉瘤病人(骨肉瘤组)、45例骨膜炎病人(骨膜炎组)和50名健康体检者(对照组)为研究对象。用RT-PCR检测血清中miR-1225-5p的表达水平。以中位数为截断值,将骨肉瘤病人分为高表达组和低表达组。绘制ROC曲线分析miR-1225-5p诊断骨肉瘤的价值;Kaplan-Meier分析和Log-Rank检验分析生存情况;采用COX比例风险回归模型分析影响病人预后的因素。 结果骨肉瘤组血清miR-1225-5p相对表达量均低于骨膜炎组和对照组(P < 0.01)。骨肉瘤病人血清miR-1225-5p表达水平与Enneking分期、肿瘤转移有关(P < 0.01和P < 0.05),而与性别、年龄、肿瘤部位、肿瘤直径和WHO骨肉瘤分类无关(P>0.05)。血清miR-1225-5p诊断骨肉瘤的曲线下面积为0.785(95%CI:0.625~0.934)。当血清miR-1225-5p相对表达量为0.46时约登指数最大,此时灵敏度和特异度分别为89.35%和71.30%。高表达组病人的总生存期和无病生存期均高于低表达组(P < 0.05)。血清miR-1225-5p是影响骨肉瘤病人OS和DFS的独立因素(P < 0.05和P < 0.01)。 结论miR-1225-5p在骨肉瘤的诊断中有重要意义,低血清miR-1225-5p表达水平与骨肉瘤病人的不良预后有关。 -
关键词:
- 骨肉瘤 /
- miR-1225-5p /
- 诊断 /
- 预后
Abstract:ObjectiveTo explore the value of serum miR-1225-5p in the diagnosis and prognosis of osteosarcoma. MethodsOne hudred and four patients with osteosarcoma, 45 patients with periostitis and 50 healthy people were divided into the osteosarcoma group, periostitis group and control group, respectively.The serum expression levels of miR-1225-5p in three groups were detected using RT-PCR.The median was set as the cutoff value, the osteosarcoma group was subdivided into the high expression group and low expression group.The value of miR-1225-5p in the diagnosis of osteosarcoma was analyzed using ROC curve, Kaplan-Meier analysis and Log-Rank test were used to analyze the survival, and the COX proportional risk regression model was used to analyze the factors affecting the prognosis of patients. ResultsThe relative expression level of serum miR-1225-5p in osteosarcoma group was lower than that in periostitis group and control group(P < 0.01).The expression of miR-1225-5p was correlated with the Enneking stage and tumor metastasis(P < 0.01 and P < 0.05), but the gender, age, tumor location, tumor diameter were not correlated with the WHO classification of osteosarcoma(P>0.05).The area under the curve of serum miR-1225-5p in the diagnosis of osteosarcoma was 0.785(95%CI: 0.625-0.934).When the relative expression of serum miR-1225-5p was 0.46, the Yoden index was the highest, and the sensitivity and specificity were 89.35% and 71.30%, respectively.The total survival time and disease-free survival time in high expression group were higher than those in low expression group(P < 0.05).The serum miR-1225-5p was an independent factor of OS and DFS in patients with osteosarcoma(P < 0.05 and P < 0.01). ConclusionsMiR-1225-5p plays an important role in the diagnosis of osteosarcoma.The low expression level of serum miR-1225-5p is related to the poor prognosis of osteosarcoma patients. -
Key words:
- osteosarcoma /
- miR-1225-5p /
- diagnosis /
- prognosis
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表 1 骨肉瘤病人血清miR-1225-5p表达水平与临床特征的关系
临床特征 高表达组
(n=52)低表达组
(n=52)χ2 P 年龄/岁 ≥18
< 1829
2326
260.35 >0.05 性别 男
女28
2430
220.16 >0.05 肿瘤部位 四肢
躯干33
1929
230.64 >0.05 Enneking分期 Ⅱ
Ⅲ27
2513
397.96 < 0.01 WHO分类 普通型骨肉瘤 27 22 6.05 >0.05 血管扩张型骨肉瘤 4 7 小细胞型骨肉瘤 2 6 骨膜骨肉瘤 1 3 骨旁骨肉瘤 8 9 髓内高分化骨肉瘤 10 5 肿瘤直径/cm < 5
≥527
2521
311.39 >0.05 远处转移 是
否19
3332
206.50 < 0.05 表 2 COX比例风险回归模型分析影响骨肉瘤病人预后的因素
因素 OS DFS HR 95%CI P HR 95%CI P Enneking分期 3.645 1.695~6.301 < 0.05 4.165 1.569~6.301 < 0.01 肿瘤转移 2.265 1.253~4.937 < 0.05 2.795 1.396~5.236 < 0.05 miR-1225-5p 3.555 1.269~5.001 < 0.05 3.934 1.568~6.152 < 0.01 -
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