中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2010年
3期
213-218
,共6页
崔杰峰%杨蓉%刘银坤%康晓楠%黄成%孙瑞霞%李杨
崔傑峰%楊蓉%劉銀坤%康曉楠%黃成%孫瑞霞%李楊
최걸봉%양용%류은곤%강효남%황성%손서하%리양
肝肿瘤%癌,肝细胞%光谱法,质量,基质辅助激光解吸电离%蛋白质陈列分析%生物学标记
肝腫瘤%癌,肝細胞%光譜法,質量,基質輔助激光解吸電離%蛋白質陳列分析%生物學標記
간종류%암,간세포%광보법,질량,기질보조격광해흡전리%단백질진렬분석%생물학표기
Liver neoplasms%Carcinoma,hepatocellular%Spectrometry,mass,matrix-assisted laser desorption ionization%Protein array analysis%Biological marker
目的 筛查潜在的小分子量低丰度的HCC血清相关候选蛋白,为肝癌复杂病理机制的全面阐明、理想早期诊断标志物的发现提供新思路.方法 分别收集81例伴有HBV感染和肝硬化背景的HCC患者血清,43例慢性乙型肝炎和36例肝硬化患者血清,采用弱阳离子交换蛋白芯片(WCX2)介质结合SELDI-TOF-MS对患者血清蛋白谱进行检测,并通过Biomarker Wizard软件对生成的蛋白指纹峰分别进行差异归类比较,然后采用消减差异分析确定HCC特异相关候选蛋白.标本检测前,对芯片检测系统的重复性进行分析.结果 同一标本同一芯片内强度CV为17.46%,m/z平均CV为0.024%,不同芯片间强度CV为17.74%,m/z平均CV为0.024%.将信噪比大于5,且超过20%标本具有的蛋白峰认定为有意义峰,在2 000～30 000 m/z范围内,共有128个蛋白指纹峰被确认.HCC与肝硬化患者血清蛋白指纹峰的差异比较分析显示,87个差异蛋白指纹峰有统计学意义(P＜0.05),其中强度差异在2倍以上的蛋白指纹峰有45个,在HCC患者血清中2倍上调的15个,2倍下调的30个.从HCC与慢性乙型肝炎血清蛋白指纹峰的差异分析发现,差异有统计学意义的蛋白指纹峰52个(P＜0.05),其中强度相差2倍以上的蛋白指纹峰有9个,HCC患者血清中2倍上调的2个,2倍下调的7个.从肝硬化与慢性乙型肝炎患者血清蛋白指纹峰的差异比较分析发现,差异有统计学意义的蛋白指纹峰79个(P＜0.05),其中强度相差2倍以上的蛋白指纹峰有28个,在肝硬化患者血清中2倍上调的17个,慢性乙型肝炎患者血清中2倍上调的11个.经消减差异分析筛查,确定在HCC患者血清下调表达的5个公共差异蛋白(2 870、3 941、2 688、3 165、5 483 m/z)及在肝硬化和HCC血清均上调表达的2个公共差异蛋白(3 588、2 017 m/z),但蛋白2 017 m/z在HCC与健康人比较的本所前期工作中的差异无统计学意义(P＞0.05).结论 由慢性乙型肝炎和肝硬化背景产生的非特异分泌蛋白的干扰,经消减分析被部分消除,获得的6个血清公共差异蛋白(2 870、3 941、2 688、3 165、5 483、3 588 m/z)在反映HCC病理特征方面具有明显的特异性,且有望成为HCC诊断中新的候选标志物.
目的 篩查潛在的小分子量低豐度的HCC血清相關候選蛋白,為肝癌複雜病理機製的全麵闡明、理想早期診斷標誌物的髮現提供新思路.方法 分彆收集81例伴有HBV感染和肝硬化揹景的HCC患者血清,43例慢性乙型肝炎和36例肝硬化患者血清,採用弱暘離子交換蛋白芯片(WCX2)介質結閤SELDI-TOF-MS對患者血清蛋白譜進行檢測,併通過Biomarker Wizard軟件對生成的蛋白指紋峰分彆進行差異歸類比較,然後採用消減差異分析確定HCC特異相關候選蛋白.標本檢測前,對芯片檢測繫統的重複性進行分析.結果 同一標本同一芯片內彊度CV為17.46%,m/z平均CV為0.024%,不同芯片間彊度CV為17.74%,m/z平均CV為0.024%.將信譟比大于5,且超過20%標本具有的蛋白峰認定為有意義峰,在2 000～30 000 m/z範圍內,共有128箇蛋白指紋峰被確認.HCC與肝硬化患者血清蛋白指紋峰的差異比較分析顯示,87箇差異蛋白指紋峰有統計學意義(P＜0.05),其中彊度差異在2倍以上的蛋白指紋峰有45箇,在HCC患者血清中2倍上調的15箇,2倍下調的30箇.從HCC與慢性乙型肝炎血清蛋白指紋峰的差異分析髮現,差異有統計學意義的蛋白指紋峰52箇(P＜0.05),其中彊度相差2倍以上的蛋白指紋峰有9箇,HCC患者血清中2倍上調的2箇,2倍下調的7箇.從肝硬化與慢性乙型肝炎患者血清蛋白指紋峰的差異比較分析髮現,差異有統計學意義的蛋白指紋峰79箇(P＜0.05),其中彊度相差2倍以上的蛋白指紋峰有28箇,在肝硬化患者血清中2倍上調的17箇,慢性乙型肝炎患者血清中2倍上調的11箇.經消減差異分析篩查,確定在HCC患者血清下調錶達的5箇公共差異蛋白(2 870、3 941、2 688、3 165、5 483 m/z)及在肝硬化和HCC血清均上調錶達的2箇公共差異蛋白(3 588、2 017 m/z),但蛋白2 017 m/z在HCC與健康人比較的本所前期工作中的差異無統計學意義(P＞0.05).結論 由慢性乙型肝炎和肝硬化揹景產生的非特異分泌蛋白的榦擾,經消減分析被部分消除,穫得的6箇血清公共差異蛋白(2 870、3 941、2 688、3 165、5 483、3 588 m/z)在反映HCC病理特徵方麵具有明顯的特異性,且有望成為HCC診斷中新的候選標誌物.
목적 사사잠재적소분자량저봉도적HCC혈청상관후선단백,위간암복잡병리궤제적전면천명、이상조기진단표지물적발현제공신사로.방법 분별수집81례반유HBV감염화간경화배경적HCC환자혈청,43례만성을형간염화36례간경화환자혈청,채용약양리자교환단백심편(WCX2)개질결합SELDI-TOF-MS대환자혈청단백보진행검측,병통과Biomarker Wizard연건대생성적단백지문봉분별진행차이귀류비교,연후채용소감차이분석학정HCC특이상관후선단백.표본검측전,대심편검측계통적중복성진행분석.결과 동일표본동일심편내강도CV위17.46%,m/z평균CV위0.024%,불동심편간강도CV위17.74%,m/z평균CV위0.024%.장신조비대우5,차초과20%표본구유적단백봉인정위유의의봉,재2 000～30 000 m/z범위내,공유128개단백지문봉피학인.HCC여간경화환자혈청단백지문봉적차이비교분석현시,87개차이단백지문봉유통계학의의(P＜0.05),기중강도차이재2배이상적단백지문봉유45개,재HCC환자혈청중2배상조적15개,2배하조적30개.종HCC여만성을형간염혈청단백지문봉적차이분석발현,차이유통계학의의적단백지문봉52개(P＜0.05),기중강도상차2배이상적단백지문봉유9개,HCC환자혈청중2배상조적2개,2배하조적7개.종간경화여만성을형간염환자혈청단백지문봉적차이비교분석발현,차이유통계학의의적단백지문봉79개(P＜0.05),기중강도상차2배이상적단백지문봉유28개,재간경화환자혈청중2배상조적17개,만성을형간염환자혈청중2배상조적11개.경소감차이분석사사,학정재HCC환자혈청하조표체적5개공공차이단백(2 870、3 941、2 688、3 165、5 483 m/z)급재간경화화HCC혈청균상조표체적2개공공차이단백(3 588、2 017 m/z),단단백2 017 m/z재HCC여건강인비교적본소전기공작중적차이무통계학의의(P＞0.05).결론 유만성을형간염화간경화배경산생적비특이분비단백적간우,경소감분석피부분소제,획득적6개혈청공공차이단백(2 870、3 941、2 688、3 165、5 483、3 588 m/z)재반영HCC병리특정방면구유명현적특이성,차유망성위HCC진단중신적후선표지물.
Objective To screen potential serum HCC associated proteins with low molecular weight and low abundance for better understanding the pathological mechanism of HCC and discovering new biomarkers.Methods All serum samples were collected from 81 HBV-related HCC patients,43 chronic hepatitis B patients and 36 cirrhosis patients.Serum protein fingerprint profiles were first generated by selected WCX2 protein chip integrating with SELDI-TOF-MS,and then normalized and aligned by Ciphergen SELDI Software 3.1.1 with Biomarker Wizard.Comparative analysis of the intensity of corresponding protein fingerprint peaks in normalized protein spectra was performed.Some protein peaks with significant difference among HCC,cirrhosis and chronic hepatitis B groups were found.The reproducibility of the SELDI system was assessed before serum protein fingerprint profiles analysis.Results The intra-and inter-assay CV for intensity and m/z in this SELDI system were 17.46% and 0.024%,and 17.74% and 0.024% respectively.Total 128 protein fingerprint peaks between 2 000 to 30 000 Da were identified under the condition of signal to noise＞5 and minimum threshold for cluster＞20%.Eighty-seven proteins were found to significantly expressed between HCC and cirrhosis groups(P＜0.05).Of the above differential proteins,forty-five proteins had changes greater than two fold,including 15 up-regulated proteins and 30 downregulated proteins in HCC sera.Between HCC and chronic hepatitis B groups,nine of fifty-two differential proteins(P＜0.05) had intensities of more than two folds,including 2 up-regulated proteins and 7 downregulated proteins in HCC sera.Between cirrhosis and chronic hepatitis B groups,twenty-eight of seventynine significantly differential proteins(P＜0.05) changed greater than two folds in intensity,including 17 up-regulated proteins in cirrhosis seru and 11 down-regulated proteins in chronic hepatitis B sera.Analysis of above leading differential proteins among three diseases using subtraction difference mode,the 5 common down-regulated proteins 2 870,3 941,2 688,3 165 and 5 483 m/z in HCC sera and 2 common up-regulated proteins 3 588 and 2 017 m/z in cirrhosis and HCC sera were screened.But no statistic difference in the level of protein 2 017m/z was found between HCC group and normal group inour previous study.Conclusion Because the interference of unspecific proteins from hepatitis B and cirrhosis could be eliminated partly in HCC sera through subtraction difference analysis,these 6 common differential proteins (2 870,3 941,2 688,3 165,5 483,3 588 m/z)have obvious advantages of increased specificity for evaluating the pathological state of HCC and might become promising candidate biomarkers in the diagnosis of HCC.
