CAJ | 학술논문

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15953、7987、9304、8948、5911，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15953、7987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15953、7987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9304、8948、5911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9304、8948、5911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5911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。揹景與目的利用蛋白質芯片技術尋找腫瘤標誌物仍是熱點，但目前跼限于僅對腎癌患者進行術前、術後的差異性蛋白質檢測。為深入檢測差異性蛋白質的變化規律，本研究探討瞭腎透明細胞癌血清差異性蛋白質錶達及臨床意義。方法選取2013年11月—2014年4月在新疆醫科大學第一附屬醫院泌尿外科手術治療併且術後經病理學專傢診斷為腎透明細胞癌患者40例為腎癌組，選擇同期在本院體檢健康誌願者以及非腎癌患者53例為對照組。運用錶麵增彊激光解析電離化飛行時間質譜繫統（SELDI - TOF - MS）和 CM10弱暘離子交換蛋白芯片技術繫統檢測腎癌組及對照組血清差異性蛋白質。應用線性支持嚮量機（SVM）方法建立蛋白質指紋圖診斷模型，留一法交扠驗證模型判彆效果。通過 ZUCI - PDAS 蛋白質譜數據分析繫統軟件對血清差異性蛋白質進行分析。結果最終5種蛋白質作為潛在標誌蛋白，質荷比（M/ Z）分彆為15953、7987、9304、8948、5911，所對應的蛋白質依次為 Bcl -2傢族細胞凋亡調節蛋白、WAP 二硫化四物覈心蛋白、Krueppel 樣因子8、單覈細胞趨化蛋白1、血清β-澱粉樣蛋白4。腎癌組術前與對照組5種蛋白質錶達水平比較，差異有統計學意義（P ﹤0.05）。腎癌組術前與術後1週、1箇月、3箇月、6箇月5種蛋白質錶達水平比較，差異均有統計學意義（P ﹤0.05）。術前5種蛋白質錶達水平均高于術後1週、1箇月、3箇月、6箇月（P ﹤0.05）；術後3、6箇月15953、7987蛋白質錶達水平與術後1週比較，差異有統計學意義（P ﹤0.05）；術後6箇月15953、7987蛋白質錶達水平與術後3箇月比較，差異有統計學意義（P ﹤0.05）。術後1、3、6箇月9304、8948、5911蛋白質錶達水平與術後1週比較，差異有統計學意義（P ﹤0.05）；術後6箇月9304、8948、5911蛋白質錶達水平與術後1箇月比較，差異有統計學意義（P ﹤0.05）；術後6箇月5911蛋白質錶達水平與術後3箇月比較，差異有統計學意義（P ﹤0.05）。5種蛋白質作為生物標誌物，預測腎透明細胞癌的靈敏度為87.5％（35/40），特異度為86.8％（46/53）。結論特異性蛋白質可能依次為 Bcl -2傢族細胞凋亡調節蛋白、WAP 二硫化四物覈心蛋白、Krueppel 樣因子8、單覈細胞趨化蛋白、血清β-澱粉樣蛋白4，共有望成為腎癌腫瘤標誌物，對腎透明細胞癌的預測價值較高，在今後腎透明細胞癌的療效評價、預後評估以及靶嚮治療方麵有一定參攷價值。
배경여목적이용단백질심편기술심조종류표지물잉시열점，단목전국한우부대신암환자진행술전、술후적차이성단백질검측。위심입검측차이성단백질적변화규률，본연구탐토료신투명세포암혈청차이성단백질표체급림상의의。방법선취2013년11월—2014년4월재신강의과대학제일부속의원비뇨외과수술치료병차술후경병이학전가진단위신투명세포암환자40례위신암조，선택동기재본원체검건강지원자이급비신암환자53례위대조조。운용표면증강격광해석전리화비행시간질보계통（SELDI - TOF - MS）화 CM10약양리자교환단백심편기술계통검측신암조급대조조혈청차이성단백질。응용선성지지향량궤（SVM）방법건립단백질지문도진단모형，류일법교차험증모형판별효과。통과 ZUCI - PDAS 단백질보수거분석계통연건대혈청차이성단백질진행분석。결과최종5충단백질작위잠재표지단백，질하비（M/ Z）분별위15953、7987、9304、8948、5911，소대응적단백질의차위 Bcl -2가족세포조망조절단백、WAP 이류화사물핵심단백、Krueppel 양인자8、단핵세포추화단백1、혈청β-정분양단백4。신암조술전여대조조5충단백질표체수평비교，차이유통계학의의（P ﹤0.05）。신암조술전여술후1주、1개월、3개월、6개월5충단백질표체수평비교，차이균유통계학의의（P ﹤0.05）。술전5충단백질표체수평균고우술후1주、1개월、3개월、6개월（P ﹤0.05）；술후3、6개월15953、7987단백질표체수평여술후1주비교，차이유통계학의의（P ﹤0.05）；술후6개월15953、7987단백질표체수평여술후3개월비교，차이유통계학의의（P ﹤0.05）。술후1、3、6개월9304、8948、5911단백질표체수평여술후1주비교，차이유통계학의의（P ﹤0.05）；술후6개월9304、8948、5911단백질표체수평여술후1개월비교，차이유통계학의의（P ﹤0.05）；술후6개월5911단백질표체수평여술후3개월비교，차이유통계학의의（P ﹤0.05）。5충단백질작위생물표지물，예측신투명세포암적령민도위87.5％（35/40），특이도위86.8％（46/53）。결론특이성단백질가능의차위 Bcl -2가족세포조망조절단백、WAP 이류화사물핵심단백、Krueppel 양인자8、단핵세포추화단백、혈청β-정분양단백4，공유망성위신암종류표지물，대신투명세포암적예측개치교고，재금후신투명세포암적료효평개、예후평고이급파향치료방면유일정삼고개치。
Background and Objective Looking for tumor markers by using protein chip technology is one of the hot topics,but many studies are still limited to detection of differentiated protein before and after surgery among patients with kidney cancer. In order to detect the variability of differentiated protein,the expression and clinical significance of serum differentiated protein among patients with clear - cell renal cell carcinoma were discussed in this study. Methods 40 patients with clear - cell renal cell carcinoma who underwent surgery in Department of Urology Surgery in the First Affiliated Hospital of Xinjiang Medical University from November 2013 to April 2014,all cases were confirmed by postoperative pathology and were included in kidney cancer group,53 healthy volunteers who underwent medical examination in the same hospital and patients without kidney cancer were included in control group. The surface - enhanced laser desorption ionization time of flight mass spectrometry(SELDI - TOF - MS)and weak cation exchange protein chip(CM10)technology systems were used to identify the serum differentiated protein in kidney cancer group and in control group. The linear support vector machine( SVM)method was used to establish the diagnostic model of protein fingerprints,and the leave - one - out cross validation was used to identify model discrimination effect. The differentially expressed proteins were analyzed by ZUCI - PDAS protein spectral data analysis system software. Results Five kinds of proteins were identified as potential marker protein,the M/ Z of these proteins was 15 953,7 987,9 304, 8 948,5 911,respectively,the corresponding protein was Bcl - 2 family apoptosis regulatory proteins,WAP four - disulfide core protein,Krueppel - like factor 8,monocyte chemotactic protein - 1,serum amyloid β - protein - 4,respectively. There were significant differences in expression levels of five kinds of proteins between kidney cancer group befrore surgery and control group(P ﹤ 0. 05). There were significant differences in expression levels of five kinds of proteins in kidney cancer group among before surgery,1 week after surgery,1 month after surgery,3 months after surgery,6 months after surgery(P ﹤ 0. 05). The expression levels of five kinds of proteins before surgery were significantly higher than those 1 week after surgery,1 month after surgery,3 months after surgery,6 months after surgery,respectively( P ﹤ 0. 05 ) . There were significant differences in expression levels of protein 15 953 and 7 987 between 3,6 months after surgery and 1 week after surgery,respectively(P ﹤0. 05). There were significant differences in expression levels of protein 15 953 and 7 987 between 3 months after surgery and 6 months after surgery( P ﹤ 0. 05). There were significant differences in expression levels of protein 9 304,8 948 and 5 911 between 1,3,6 months after surgery and 1 week after surgery,respectively(P ﹤ 0. 05). There was significant difference in expression levels of protein 5 911 between 3 months after surgery and 6 months after surgery( P ﹤ 0. 05). The sensitivity and specificity of five kinds of proteins in predicting clear - cell renal cell carcinoma was 87. 5 % (35 / 40)and 86. 8% (46 / 53), respectively. Conclusion The specific proteins may include Bcl - 2 family apoptosis regulatory proteins,WAP four - disulfide core protein,Krueppel - like factor 8,monocyte chemotactic protein - 1 and serum amyloid β - protein - 4,respectively, which may be used as tumor markers of kidney cancer,these proteins have high predictive value for clear - cell renal cell carcinoma,and may contribute to therapeutic evaluation,prognosis and targeted therapy for clear - cell renal cell carcinoma.