中国临床新医学
中國臨床新醫學
중국림상신의학
CHINESE JOURNAL OF NEW CLINICAL MEDICINE
2015年
5期
394-397
,共4页
施强%牟云%于大海%黎明武%卿海云%李晶
施彊%牟雲%于大海%黎明武%卿海雲%李晶
시강%모운%우대해%려명무%경해운%리정
小鼠%播散肿瘤细胞%激光捕获显微切割(LCM)%RB1CC1%纯合性缺失
小鼠%播散腫瘤細胞%激光捕穫顯微切割(LCM)%RB1CC1%純閤性缺失
소서%파산종류세포%격광포획현미절할(LCM)%RB1CC1%순합성결실
Mouse%Disseminated tumor cells%LCM%RB1CC1 gene%Homozygous deletion
目的:初步探讨小鼠口腔癌模型重度异常增生时期骨髓播散细胞RB1 CC1基因纯合性缺失及其意义。方法应用激光捕获显微切割( LCM)技术捕获35张重度异常增生时期小鼠骨髓单个核细胞涂片的单个细胞角蛋白阳性( CK+)细胞,单细胞全基因组扩增技术( WGA)扩增其DNA,PCR检测其视网膜母细胞瘤诱导卷曲蛋白( RB1CC1)的纯合性缺失情况,并与舌部正常、重度异常增生以及鳞癌组织各4例对照。结果 LCM成功捕获CK+细胞35个,WGA成功扩增3个,RB 1CC1纯合性缺失情况分别为单个CK+细胞(2/3),舌癌变组织(2/4),舌部正常组织(0/4),重度异常增生组织(0/4)。结论小鼠口腔癌模型重度异常增生时期骨髓CK+细胞有RB1CC1基因纯合性缺失,该时期骨髓CK+细胞可能是播散肿瘤细胞。
目的:初步探討小鼠口腔癌模型重度異常增生時期骨髓播散細胞RB1 CC1基因純閤性缺失及其意義。方法應用激光捕穫顯微切割( LCM)技術捕穫35張重度異常增生時期小鼠骨髓單箇覈細胞塗片的單箇細胞角蛋白暘性( CK+)細胞,單細胞全基因組擴增技術( WGA)擴增其DNA,PCR檢測其視網膜母細胞瘤誘導捲麯蛋白( RB1CC1)的純閤性缺失情況,併與舌部正常、重度異常增生以及鱗癌組織各4例對照。結果 LCM成功捕穫CK+細胞35箇,WGA成功擴增3箇,RB 1CC1純閤性缺失情況分彆為單箇CK+細胞(2/3),舌癌變組織(2/4),舌部正常組織(0/4),重度異常增生組織(0/4)。結論小鼠口腔癌模型重度異常增生時期骨髓CK+細胞有RB1CC1基因純閤性缺失,該時期骨髓CK+細胞可能是播散腫瘤細胞。
목적:초보탐토소서구강암모형중도이상증생시기골수파산세포RB1 CC1기인순합성결실급기의의。방법응용격광포획현미절할( LCM)기술포획35장중도이상증생시기소서골수단개핵세포도편적단개세포각단백양성( CK+)세포,단세포전기인조확증기술( WGA)확증기DNA,PCR검측기시망막모세포류유도권곡단백( RB1CC1)적순합성결실정황,병여설부정상、중도이상증생이급린암조직각4례대조。결과 LCM성공포획CK+세포35개,WGA성공확증3개,RB 1CC1순합성결실정황분별위단개CK+세포(2/3),설암변조직(2/4),설부정상조직(0/4),중도이상증생조직(0/4)。결론소서구강암모형중도이상증생시기골수CK+세포유RB1CC1기인순합성결실,해시기골수CK+세포가능시파산종류세포。
Objective To observe the homozygous deletions of RB1CC1 gene of disseminated cells in bone marrow of mouse oral carcinoma model in the severe abnormal hyperplasia stage.Methods The cytokeratin positive ( CK+) cells from 35 single nuclear cells smear of bone marrow of mouse oral carcinoma model in the severe abnor-mal hyperplasia stage were isolated by Laser Capture Microdissection ( LCM) .The DNA of a single cell was ampli-fied by the technique of single-cell Whole Genome Amplification (WGA) and the homozygous deletions of RB1CC1 gene of the DTC were detected by Polymerase Chain Reaction ( PCR) .Four normal, 4 severe abnormal hyperplasia and 4 squamous cell carcinoma tongue were taken as control.Results Thirty five CK+cells from the 35 smear re-spectively were successfully isolated by LCM.Genomic DNA were successfully amplified by single-cell WGA in 3 out of 35 cells.Homozygous deletions of RB1CC1 gene were found in 2 of the 3 CK+cells and 2 in 4 cases of squamous cell carcinoma tongue mucosa.However, the deletion was not found in the rest tissues.Conclusion Homozygous deletion of RB1CC1 gene of the CK+positive cells in the severe abnormal hyperplasia stage may be the disseminated tumor cells.