中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2011年
12期
898-903
,共6页
癌,肝细胞%肿瘤转移%糖基化%CD44v6%DNA甲基化
癌,肝細胞%腫瘤轉移%糖基化%CD44v6%DNA甲基化
암,간세포%종류전이%당기화%CD44v6%DNA갑기화
Carcinoma,hepatocellular%Neoplasm metastasis%Glycosylation%CD44v6%DNA methlation
目的 探讨CD44S及其变异体CD44v分子的表达和其糖基化与肝癌细胞侵袭转移的关系. 方法 用免疫组织化学法、量子点、RT-PCR、Western blot、细胞免疫荧光染色、甲基化特异性聚合酶链反应等技术检测转移与非转移性肝癌组织、不同转移潜能人肝癌细胞株中CD44S及其变异体CD44v的定位与表达;并应用多重凝集素印迹法检测这些细胞株中CD44v6的糖基化差异.组间均数比较应用方差分析及t检验,多组间等级资料的比较采用Wilcoxon秩和检验,各组间率的比较采用x2检验.结果 免疫组织化学结果显示,CD44S蛋白定位以细胞质为主; CD44v3、CD44v4/5蛋白定位于细胞膜与细胞质；而CD44v6主要定位于细胞膜.组织芯片结果显示,相对于CD44S及其他CD44v,CD44v6在转移组的表达水平高于非转移组(阳性率为75％与46％),差异具有统计学意义(x2=8.828,P＜0.05);量子点检测(t=2.392,P＜0.05)与血清学检测(t=2.56,P＜0.05)也证实这一结果.Western blot结果显示,CD44v6的表达与肝癌细胞转移潜能呈正相关.此外,在MHCC97L、MHCC97H肝癌细胞中CD44v6基因启动子发生不完全甲基化,而在Hep3B细胞中则发生完全甲基化.而且,相对于Hep3B细胞,MHCC97L及MHCC97H细胞中CD44v6蛋白对朝鲜槐凝集素、黑接骨木凝集素及麦胚凝激素的亲和力较高. 结论 在CD44S及其变异体CD44v中,CD44v6蛋白的高表达与肝癌转移潜能的增强呈正相关；其高表达可能与基因启动子低甲基化有关.此外,CD44v6蛋白唾液酸寡糖链的增加可能与肝癌细胞转移潜能增高有关.
目的 探討CD44S及其變異體CD44v分子的錶達和其糖基化與肝癌細胞侵襲轉移的關繫. 方法 用免疫組織化學法、量子點、RT-PCR、Western blot、細胞免疫熒光染色、甲基化特異性聚閤酶鏈反應等技術檢測轉移與非轉移性肝癌組織、不同轉移潛能人肝癌細胞株中CD44S及其變異體CD44v的定位與錶達;併應用多重凝集素印跡法檢測這些細胞株中CD44v6的糖基化差異.組間均數比較應用方差分析及t檢驗,多組間等級資料的比較採用Wilcoxon秩和檢驗,各組間率的比較採用x2檢驗.結果 免疫組織化學結果顯示,CD44S蛋白定位以細胞質為主; CD44v3、CD44v4/5蛋白定位于細胞膜與細胞質；而CD44v6主要定位于細胞膜.組織芯片結果顯示,相對于CD44S及其他CD44v,CD44v6在轉移組的錶達水平高于非轉移組(暘性率為75％與46％),差異具有統計學意義(x2=8.828,P＜0.05);量子點檢測(t=2.392,P＜0.05)與血清學檢測(t=2.56,P＜0.05)也證實這一結果.Western blot結果顯示,CD44v6的錶達與肝癌細胞轉移潛能呈正相關.此外,在MHCC97L、MHCC97H肝癌細胞中CD44v6基因啟動子髮生不完全甲基化,而在Hep3B細胞中則髮生完全甲基化.而且,相對于Hep3B細胞,MHCC97L及MHCC97H細胞中CD44v6蛋白對朝鮮槐凝集素、黑接骨木凝集素及麥胚凝激素的親和力較高. 結論 在CD44S及其變異體CD44v中,CD44v6蛋白的高錶達與肝癌轉移潛能的增彊呈正相關；其高錶達可能與基因啟動子低甲基化有關.此外,CD44v6蛋白唾液痠寡糖鏈的增加可能與肝癌細胞轉移潛能增高有關.
목적 탐토CD44S급기변이체CD44v분자적표체화기당기화여간암세포침습전이적관계. 방법 용면역조직화학법、양자점、RT-PCR、Western blot、세포면역형광염색、갑기화특이성취합매련반응등기술검측전이여비전이성간암조직、불동전이잠능인간암세포주중CD44S급기변이체CD44v적정위여표체;병응용다중응집소인적법검측저사세포주중CD44v6적당기화차이.조간균수비교응용방차분석급t검험,다조간등급자료적비교채용Wilcoxon질화검험,각조간솔적비교채용x2검험.결과 면역조직화학결과현시,CD44S단백정위이세포질위주; CD44v3、CD44v4/5단백정위우세포막여세포질；이CD44v6주요정위우세포막.조직심편결과현시,상대우CD44S급기타CD44v,CD44v6재전이조적표체수평고우비전이조(양성솔위75％여46％),차이구유통계학의의(x2=8.828,P＜0.05);양자점검측(t=2.392,P＜0.05)여혈청학검측(t=2.56,P＜0.05)야증실저일결과.Western blot결과현시,CD44v6적표체여간암세포전이잠능정정상관.차외,재MHCC97L、MHCC97H간암세포중CD44v6기인계동자발생불완전갑기화,이재Hep3B세포중칙발생완전갑기화.이차,상대우Hep3B세포,MHCC97L급MHCC97H세포중CD44v6단백대조선괴응집소、흑접골목응집소급맥배응격소적친화력교고. 결론 재CD44S급기변이체CD44v중,CD44v6단백적고표체여간암전이잠능적증강정정상관；기고표체가능여기인계동자저갑기화유관.차외,CD44v6단백타액산과당련적증가가능여간암세포전이잠능증고유관.
Objective To explore the relationship of CD44 expression or glycosylation and hepatocellular carcinoma (HCC) metastasis.Methods IHC,Quantum dots dtection,RT-PCR,Western blot,Cellular immune fluorescence and MS-PCR were used to identify CD44 expression in HCC samples and a series of human HCC cell lines with different metastatic potentials.Lectin array was used to reveal the relationship of CD44v6 glycosylation and human HCC metastasis.Results Immunohistochemistry analysis showed that CD44v6 was mainly distributed on the cell membrane,while CD44S immunoreactivity was prominently in the cytoplasm,CD44v3 and CD44v4/5 were in cytoplasm on membrane.Among CD44S and those CD44 variants,only the expression of CD44v6 was higher in metastasis HCC samples as compared to that in non-metastasis group (x2 =8.828,P ＜ 0.05).This result was also re-confirmed by the result of Quantum dots(t=2.392,P＜0.05) and serum detection (t=2.56,P＜0.05).We found completely methylation of CD44v6 gene in Hep3B and incomplete methylation in MHCC97H and MHCC97L cell lines with metastatic potentials.The lectin affinity assay indicated that lectin MAL,SNA and WGA showed more affinity to MHCC97H and MHCC97Lcell lines than that of the non-metastatic Hep3B cell lines.Conclusions CD44v6 over-expression presents a positive correlation with HCC metastatic potential,which may be associated with DNA methyhylation level in promoter sequence.The increasing sialic acid modified glycan of CD44v6 might be related to HCC metastatic ability.
