miR-1237-3p和miR-140-3p的异常表达对脊柱脊索瘤预后影响的病例对照研究

发布时间：2018-06-26 04:50

本文选题：microRNA + 脊柱　；参考：《中南大学》2014年硕士论文

【摘要】：目的：探讨脊柱脊索瘤中miR-1237-3p和miR-140-3p的表达对患者预后的影响。方法：利用Agilent人类miRNA芯片技术(微阵列技术),回顾性分析脊索瘤组织和髓核组织(作为阴性对照)中miRNA的表达情况。根据微阵列分析结果,使用qRT-PCR技术对来自42名脊索瘤患者(男性27名和女性15名,年龄29-76岁,平均52.93±11.50岁)的肿瘤组织和14名腰椎间盘突出症患者(作为健康对照组,男性8名和女性6名,年龄24-73岁,平均49.21±13.35岁)的髓核组织中差异表达的miRNAs(miR-1237-3p和miR-140-3p)进行定量检测。所有样本均采取手术方式获得。同时获得患者以下的临床及病理学资料,包括年龄、性别、肿瘤大小、肿瘤部位、周围肌肉组织浸润和术前复发状态。病例组42例患者(原发性患者32名,复发性患者10名)根据miR-1237-3p和miR-140-3p平均表达水平(分别为0.18±0.27和3.56±3.17)分为两组。探讨上述两个miRNA分子的相对表达量与脊索瘤患者临床病理学特征的关系。利用Kaplan-Meier法估计不同的miR-1237-3p和miR-140-3p表达组患者间的生存率和生存曲线,并采用Cox比例风险模型分析多种因素对生存时间的影响。术后根据患者的临床和影像学表现进行随访。无复发生存时间(Recurrence-free survival time, RFS)定义为从肿瘤手术切除到根据临床检查和影像学评估出现疾病复发的时间。在进行该次手术治疗之前,没有患者曾接受化疗或放射治疗。结果： 1.病例组和对照组中性别构成和年龄分布差异无统计学意义(χ2=0.229,P=0.633和t=1.005,P=0.319)。 2.微阵列分析结果显示,相比对照组髓核组织,有182个miRNA在脊索瘤中存在差异表达(Fold change≥2.0且P≤0.05),其中,110个miRNA为高表达,另72个miRNA呈低表达状态。miR-1237-3p和miR-140-3p在脊索瘤组织中存在表达异常(P=0.0036,fold change:12.62和P=0.0211,fold change=19.76).该结果为后续qRT-PCR实验进一步证实,其结果表明miR-1237-3p在脊索瘤组织中的表达水平低于髓核组织(t=-2.906,P=0.011),而miR-140-3p在脊索瘤组织中却呈高表达状态(t=3.530,P=0.001). 3. miR-1237-3p低表达患者较高表达患者出现周围肌肉组织浸润的风险越高(OR=6.000,95%CI:1.472-24.454,P=0.007)。而miR-140-3p高表达组患者较低表达组患者出现周围肌肉组织浸润的风险越高(OR=3.238,95%CI:1.719-6.101,P=0.000)。 4.随访时间为5-72个月。在随访期内,有32例(76.19%)患者出现复发。中位生存时间为28.0个月(5-72个月)。Kapan-meier生存分析显示术前伴周围肌肉组织浸润的患者RFS时间较无周围肌肉组织浸润患者短(χ2=26.917,P=0.000,log-rank检验)。miR-1237-3p低表达组患者RFS时间较高表达组患者短(χ2=16.217, P=0.00,log-rank检验),而miR-140-3p高表达组患者RFS时间较低表达组患者短(χ2=31.270,P=0.000,log-rank检验)。 5.在多因素Cox回归模型中,有三个因素进入模型,分别为：miR-1237-3p和miR-140-3p的表达(HR=0.001,95%CI:0.000-0.141,P=0.006和HR=1.267,95%CI:1.055-1.522, P=0.011)和周围肌肉组织浸润(HR=4.759,95%CI:1.294-17.510, P=0.019). 结论：脊柱脊索瘤中miR-1237-3p和miR-140-3p的异常表达与肿瘤复发和浸润有关,提示miR-1237-3p和miR-140-3p可作为脊柱脊索瘤患者复发和预后的预测指标。
[Abstract]:Objective: To investigate the effect of miR-1237-3p and miR-140-3p expression on prognosis of spinal chordoma.
Methods: the expression of miRNA in the chordoma tissue and nucleus pulposus (as negative control) was retrospectively analyzed using the Agilent human miRNA chip technique (microarray technique). According to the results of microarray analysis, qRT-PCR was used to detect the swelling of 42 patients (27 men and 15 women, 29-76 years old and 52.93 + 11.50 years old). The differential expression of miRNAs (miR-1237-3p and miR-140-3p) in the nucleus pulposus of the tumor tissue and 14 lumbar intervertebral disc herniation patients (8 men and 6 men, 24-73 years old and 49.21 + 13.35 years old) was quantified. All the samples were obtained by surgical procedure. The clinical and pathological features of the patients were also obtained. Data, including age, sex, tumor size, tumor site, peripheral muscle tissue infiltration and preoperative recurrence. 42 patients (32 of primary patients and 10 recurrent patients) were divided into two groups based on the average expression level of miR-1237-3p and miR-140-3p (0.18 + 0.27 and 3.56 + 3.17 respectively). The relative tables of the two miRNA molecules mentioned above were discussed. The relationship between DA and the clinicopathological features of the patients with chordoma. The Kaplan-Meier method was used to estimate the survival rate and survival curve of patients with different miR-1237-3p and miR-140-3p expression groups, and the Cox proportional hazard model was used to analyze the impact of a variety of factors on the survival time. Recurrence-free survival time (RFS) is defined as the time for the recurrence of the disease from the resection of the tumor to the clinical and imaging evaluation. No patient has received chemotherapy or radiation before this operation.
Result:
1. there was no significant difference in sex composition and age distribution between case group and control group (x 2=0.229, P=0.633 and t=1.005, P=0.319).
2. microarray analysis showed that there were 182 different expressions of miRNA in chordoma compared with the control group (Fold change > 2 and P < 0.05), of which 110 miRNA were highly expressed and 72 miRNA showed low expression of.MiR-1237-3p and miR-140-3p in chordoma tissues (P=0.0036, fold change:12.62 and P=0.021). 1, fold change=19.76). The results were further confirmed by subsequent qRT-PCR experiments. The results showed that the expression level of miR-1237-3p in the chordoma tissue was lower than that of the nucleus pulposus (t=-2.906, P=0.011), but the miR-140-3p in the chordoma tissue was highly expressed (t=3.530, P= 0.001).
The higher risk of peripheral muscle tissue infiltration in patients with low expression of 3. miR-1237-3p was higher (OR=6.000,95%CI:1.472-24.454, P=0.007). The higher risk of peripheral muscle tissue infiltration in patients with low expression of miR-140-3p was higher (OR=3.238,95%CI: 1.719-6.101, P=0.000).
4. the duration of follow-up was 5-72 months. During the follow-up period, 32 patients (76.19%) had a recurrence. The median survival time was 28 months (5-72 months).Kapan-meier survival analysis showed that the RFS time of patients with infiltration of peripheral muscle tissue was shorter than those without peripheral muscle tissue infiltration (x 2=26.917, P=0.000, log-rank test) low expression of.MiR-1237-3p. The patients in the group with high expression of RFS were short (x 2=16.217, P=0.00, log-rank test), while the miR-140-3p high expression group had shorter RFS time than the lower expression group (x 2=31.270, P=0.000, log-rank test).
5. in the multifactor Cox regression model, three factors entered the model, miR-1237-3p and miR-140-3p (HR=0.001,95%CI:0.000-0.141, P=0.006 and HR=1.267,95%CI:1.055-1.522, P=0.011) and the infiltration of peripheral muscle tissue (HR=4.759,95%CI:1.294-17.510, P= 0.019).
Conclusion: the abnormal expression of miR-1237-3p and miR-140-3p in spinal chordoma is associated with tumor recurrence and invasion, suggesting that miR-1237-3p and miR-140-3p can be used as a predictor of recurrence and prognosis in spinal chordoma.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.4

【共引文献】