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背景与目的利用蛋白质芯片技术寻找肿瘤标志物仍是热点,但目前局限于仅对肾癌患者进行术前、术后的差异性蛋白质检测。为深入检测差异性蛋白质的变化规律,本研究探讨了肾透明细胞癌血清差异性蛋白质表达及临床意义。方法选取2013年11月—2014年4月在新疆医科大学第一附属医院泌尿外科手术治疗并且术后经病理学专家诊断为肾透明细胞癌患者40例为肾癌组,选择同期在本院体检健康志愿者以及非肾癌患者53例为对照组。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。结果最终5种蛋白质作为潜在标志蛋白,质荷比(M/Z)分别为15 953、7 987、9 304、8 948、5 911,所对应的蛋白质依次为Bcl-2家族细胞凋亡调节蛋白、WAP二硫化四物核心蛋白、Krueppel样因子8、单核细胞趋化蛋白1、血清β-淀粉样蛋白4。肾癌组术前与对照组5种蛋白质表达水平比较,差异有统计学意义(P<0.05)。肾癌组术前与术后1周、1个月、3个月、6个月5种蛋白质表达水平比较,差异均有统计学意义(P<0.05)。术前5种蛋白质表达水平均高于术后1周、1个月、3个月、6个月(P<0.05);术后3、6个月15 953、7 987蛋白质表达水平与术后1周比较,差异有统计学意义(P<0.05);术后6个月15 953、7 987蛋白质表达水平与术后3个月比较,差异有统计学意义(P<0.05)。术后1、3、6个月9 304、8 948、5 911蛋白质表达水平与术后1周比较,差异有统计学意义(P<0.05);术后6个月9 304、8 948、5 911蛋白质表达水平与术后1个月比较,差异有统计学意义(P<0.05);术后6个月5 911蛋白质表达水平与术后3个月比较,差异有统计学意义(P<0.05)。5种蛋白质作为生物标志物,预测肾透明细胞癌的灵敏度为87.5%(35/40),特异度为86.8%(46/53)。结论特异性蛋白质可能依次为Bcl-2家族细胞凋亡调节蛋白、WAP二硫化四物核心蛋白、Krueppel样因子8、单核细胞趋化蛋白、血清β-淀粉样蛋白4,共有望成为肾癌肿瘤标志物,对肾透明细胞癌的预测价值较高,在今后肾透明细胞癌的疗效评价、预后评估以及靶向治疗方面有一定参考价值。
Background and Objective It is still a hot spot to find tumor markers by using protein chip technology, but it is currently limited to preoperative and postoperative differential protein detection in patients with renal cell carcinoma. In order to further examine the variation of differential proteins, this study explored the serum differential protein expression in renal clear cell carcinoma and its clinical significance. Methods From November 2013 to April 2014, 40 cases of renal clear cell carcinoma were diagnosed as renal cell carcinoma by operation of urology in the First Affiliated Hospital of Xinjiang Medical University. 53 healthy volunteers and non-renal cancer patients as control group. SELDI-TOF-MS and CM10 weak cation exchange protein chip were used to detect serum differential proteins in RCC and control groups by surface enhanced laser desorption / ionization time of flight mass spectrometry (SELDI-TOF-MS). The diagnosis model of protein fingerprinting was established by using the method of linear support vector machine (SVM), and the discriminant effect was verified by a cross-validation model. Serum differential proteins were analyzed by ZUCI-PDAS protein profiling system software. Results As the potential marker proteins, the mass-to-charge ratios (M / Z) of the final five proteins were 15 953, 7 987, 9 304, 8 948 and 5 911 respectively. The corresponding proteins were, in turn, Bcl-2 family apoptosis regulatory proteins , WAP disulfide core protein, Krueppel-like factor 8, monocyte chemotactic protein 1, serum beta-amyloid 4. There were significant differences in the expression levels of five proteins between preoperative and control groups (P <0.05). There were significant differences in the expression of five proteins between preoperative and postoperative 1 week, 1 month, 3 months and 6 months in renal cell carcinoma (P <0.05). The levels of protein expression in the five preoperative groups were significantly higher than those at 1 week, 1 month, 3 months and 6 months after operation (P <0.05). The protein expression levels of 15 953 and 7 987 at 3 and 6 months postoperatively were significantly different from those after operation (P <0.05). The protein expression level of 15 953,7 987 at 6 months after operation was significantly higher than that at 3 months after operation (P <0.05). The protein expression levels of 9 304,8 948,5 911 at 1, 3 and 6 months after operation were significantly different from those at 1 week after operation (P <0.05); at 6 months after operation 9 304,898,458,5 (P <0.05). The protein expression level of 5 911 at 6 months after operation was significantly higher than that at 3 months after operation (P <0.05) ). The five proteins as biomarkers were 87.5% (35/40) in sensitivity and 86.8% (46/53) in predicting renal clear cell carcinoma. CONCLUSIONS: The specific proteins may be Bcl-2 family members regulatory proteins, WAP disulfide core protein, Krueppel-like factor 8, monocyte chemoattractant protein, and serum beta-amyloid protein 4, Tumor markers, high predictive value of renal clear cell carcinoma, renal cell carcinoma in the future evaluation of the efficacy, prognosis and targeted therapy have some reference value.