Diagnostic value of promoter methylation and protein expression of plasma RNF180 gene in gastric cancer
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摘要:
目的 探讨血浆RNF180基因启动子甲基化及其蛋白表达在胃癌临床诊断中的价值。 方法 采用甲基化特异性PCR法(MSP)和ELISA法检测各组患者血浆中RNF180甲基化状态和RNF180蛋白表达情况，并分别将其与各临床病理指标进行统计学分析。 结果 MSP结果显示胃癌患者血浆中RNF180基因启动子甲基化率为62.75%，健康者为21.88%，其差异具有统计学意义(P < 0.01)。RNF180甲基化阳性率与肿瘤大小、临床分期、分化程度、淋巴结转移以及远处转移密切相关(P < 0.05)。ELISA结果显示胃癌血浆RNF180蛋白水平(23.22±1.36) μg/mL明显低于对照健康组(34.25±2.44) μg/mL，差异具有统计学意义(P < 0.01)。胃癌血浆RNF180蛋白表达水平与临床病理特征无统计学意义(P>0.05)。 结论 胃癌患者血浆RNF180基因表达存在高甲基化率，而RNF180蛋白表达明显下降，提示血浆RNF180基因甲基化可作为一种微创的检测方法在临床胃癌诊断中有潜在的应用价值。 Abstract:Objective To investigate the diagnostic value of the promoter methylation of plasma RNF180 gene and its protein expression for the detection of gastric cancer. Methods Methylation-specific polymerase-chain reaction (MSP) and enzyme-linked immuno-sorbent assay (ELISA) were performed to detect DNA methylation and protein expression of the RNF180 gene, respectively. The correlations of DNA methylation and protein expression of the RNF180 gene with the clinico-pathological parameters of gastric carcinoma were then separately analyzed. Results MSP showed that the methylation rates of the RNF180 gene were 62.75% and 21.88% in the plasma of patients with gastric carcinoma and healthy volunteers, respectively; this result indicated that the two groups significantly differed (P < 0.01). The methylation of the RNF180 gene was associated with tumor size, clinical stage, tumor differentiation, lymph node metastasis, and distant metastasis (P < 0.05). ELISA results showed that the protein expression of the RNF180 gene [(23.22 ± 1.36) μg/mL] was significantly lower (P < 0.01) in the plasma of patients with gastric carcinoma than in the plasma of healthy volunteers [(34.25 ± 2.44) μg/mL]. However, the protein expression of the RNF180 gene was not associated with clinicopathological parameters (P>0.05). Conclusion The RNF180 gene is expressed at a hypermethylation rate, and the corresponding protein expression level is decreased in the plasma of individuals with gastric carcinoma. Therefore, RNF180 gene methylation in plasma could be applied to detect microinvasion for the clinical diagnosis of gastric cancer. -
Key words:
- RNF180 gene /
- methylation /
- gastric cancer /
- plasma /
- methylation-specific polymerase-chain reaction
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图 1 正常健康者与胃癌患者血浆RNF180基因MSP电泳图
Figure 1. MSP electrophoretograms of aberrant RNF180 methylation in gastric cancer patients and healthy controls
M: methylation, U: no methylation. Upper panel: plasma samples of the normal controls in cases 1 to 8. Lower panel: Plasma samples of the gastric cancer patients in cases 1 to 8
图 2 正常和胃癌患者血浆RNF180甲基化发生率(P < 0.01)
Figure 2. Frequency of detecting methylated RNF180 DNA in the plasma of patients with gastric cancer and normal controls (P < 0.01)
图 3 正常健康者和胃癌患者血浆RNF180蛋白水平表达
Figure 3. ELISA results of RNF180 protein levels in patients with gastric cancer and the controls subjects
表 1 RNF180 MSP甲基化引物序列
Table 1. Primer sequences of RNF180 in MSP
表 2 胃癌患者血浆RNF180基因甲基化率与其临床病理特征相关性
Table 2. Correlation of RNF180 methylation with clinicopathological features in plasma samples of gastric cancer patients
表 3 胃癌患者血浆RNF180蛋白表达与临床病理特征相关性
Table 3. Correlation of RNF180 protein levels in plasma samples of gastric cancer patients with chinicopathologic features
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