富组蛋白-3在肾透明细胞癌患者中的表达及临床意义

Expression and clinical significance of Histatin-3 in clear cell renal cell carcinoma

目的利用蛋白质芯片技术筛查肾透明细胞癌患者手术前后血清差异性表达的蛋白质,并探讨其临床意义。方法选取2015年10月至2016年10月在新疆医科大学第一附属医院泌尿外科手术治疗并经术后病理证实为肾透明细胞癌患者89例为肾癌组,选择同期在我院体检的健康志愿者以及非肾细胞癌患者100例为对照组。肾癌组术后规律随访1年。运用表面增强激光解析电离化飞行时间质谱系统(SELDI-TOF-MS)和CM10弱阳离子交换蛋白芯片技术系统检测肾癌组及对照组血清差异性表达的蛋白质。应用线性支持向量机(SVM)方法建立蛋白质指纹图诊断模型,留一法交叉验证模型判别效果。通过ZUCI-PDAS蛋白质谱数据分析系统软件对血清差异性蛋白质进行分析。结果质荷比(M/Z)为2745的蛋白质在肾癌组与对照组呈差异性表达,所对应为富组蛋白-3,其表达水平在肾癌术前组较对照组呈低表达,差异有统计学意义(P<0.05);肾癌组术前与术后1个月、3个月、6个月以及12个月后的表达水平比较,术前表达水平低于术后各随访段水平,差异均有统计学意义(P<0.05)。术后3、6、12个月表达水平与术后1个月相比较,差异有统计学意义(P<0.05)。术后6、12个月与术后3个月表达水平相比较,差异无统计学意义(P>0.05)。依据富组蛋白-3在肾癌组患者与健康人血清中表达量不同,利用ROC曲线法分析,曲线下面积AUC=0.804,灵敏度为86.7%,特异度为66.7%。该蛋白质表达强度的某一界点作为标记,预测诊断肾透明细胞癌的灵敏度为86.3%(76/89),特异度为88.0%(88/100)。结论富组蛋白-3在肾癌与健康人群中的表达存在差异,且肾癌术后其表达水平升高,提示富组蛋白-3与肾细胞癌具有相关性,有望成为肾透明细胞癌新的分子标记物。

Objective To screen differentially expressed proteins in serum of patients with renal clear cell carcinoma(RCC) before and after operation by protein chip technique and to explore its clinical significance. Method From October 2015 to October 2016, 89 patients with renal clear cell carcinoma(RCC) who underwent urological surgery in the First Affiliated Hospital of Xinjiang Medical University were selected as the RCC group, and 100 healthy volunteers and non-RCC patients were selected as the control group. The RCC group was followed up for 1 year after operation. Serum differential proteins were detected by surface enhanced laser time of flight mass spectrometry(SELDI-TOF-MS) and CM10 weak cation exchange protein chip. Linear Support Vector Machine(SVM) method was used to establish a protein fingerprint diagnosis model, leaving a cross validation model to identify the effect. The serum differential proteins were analyzed by ZUCI-PDAS protein mass spectrometry data analysis system. Results The protein of mass-charge ratio(M/Z) 2745 was differentially expressed between RCC group and control group, and corresponding protein is Histatin-3, its expression level was lower in RCC group than that in control group before operation(P<0.05). The preoperative expression level of renal cell carcinoma group was lower than that of the follow-up group 1 month, 3 months, 6 months and 12 months after operation(P<0.05). The expression level at 3, 6, 12 months after operation was significantly higher than that at 1 month after operation(P<0.05);There was no significant difference between 6 months, 12 months after operation and 3 months after operation(P>0.05). According to the different expression levels of Histone-3 in the two groups, ROC curve analysis showed that the area under the curve(AUC) was 0.804, sensitivity was 86.7%, specificity was 66.7%. A certain boundary of protein expression intensity was used as a marker, the sensitivity of predicting renal clear cell carcinoma was 86.3%(76/89), and the specificity was 88%(88/100). Conclusion The expression of Histone-3 in renal cell carcinoma was different from that in healthy people, and the expression level of Histone-3 increased after operation, suggesting that Histone-3 is correlated with renal cell carcinoma and may become a new molecular marker of renal clear cell carcinoma.