CAJ | 학술논문

目的从基因水平了解肝细胞癌(HCC)内的星状细胞(HSC)与正常肝HSC的异同.方法密度梯度离心分离HSC,用大鼠HCC细胞株CSF的条件培养基诱导HSC活化.cDNA微阵列比较静止、体外培养活化和诱导活化HSC之间22 012个基因的表达,并通过实时逆转录聚合酶链反应(real-time RT-PCR)和Western blot检测进行验证.结果与静止HSC相比,体外培养HSC共有1672个基因差异表达,包括促炎症因子、细胞表面受体、黏附分子、信号转导分子、免疫因子等,其中1012个基因上调,660个基因下调.肿瘤诱导活化HSC有711个基因差异表达,其中420个基因上调,291个基因下调,有一部分基因表达与体外培养相同,部分基因如Raf1、Rac2、Adam17、Wnt6、MMP-9和TNF等,呈特异性表达.real-time RT-PCR和Western blot检测结果与cDNA微阵列检测结果相符.结论 HCC细胞可特异性驱动HSC的活化,诱导活化的HSC在HCC细胞的浸润和转移中可能起重要作用.瘤内活化的HSC应作为研究HSC生物学功能的标准.
목적 종기인수평료해간세포암(HCC)내적성상세포(HSC)여정상간HSC적이동.방법 밀도제도리심분리HSC,용대서HCC세포주CSF적조건배양기유도HSC활화.cDNA미진렬비교정지、체외배양활화화유도활화HSC지간22 012개기인적표체,병통과실시역전록취합매련반응(real-time RT-PCR)화Western blot검측진행험증.결과 여정지HSC상비,체외배양HSC공유1672개기인차이표체,포괄촉염증인자、세포표면수체、점부분자、신호전도분자、면역인자등,기중1012개기인상조,660개기인하조.종류유도활화HSC유711개기인차이표체,기중420개기인상조,291개기인하조,유일부분기인표체여체외배양상동,부분기인여Raf1、Rac2、Adam17、Wnt6、MMP-9화TNF등,정특이성표체.real-time RT-PCR화Western blot검측결과여cDNA미진렬검측결과상부.결론 HCC세포가특이성구동HSC적활화,유도활화적HSC재HCC세포적침윤화전이중가능기중요작용.류내활화적HSC응작위연구HSC생물학공능적표준.
Objective Hepatic stellate cells (HSC) in hepatocellular carcinoma (HCC) transdifferentiate into extracellular matrix-producing myofibroblasts. Activated HSC can promote invasion and metastasis of HCC. To understand the differences of HSC in normal liver and HCC, we compared the gene expression patterns in HCC cell induction-actlvated and culture-activated rat HSC. Methods HSC were isolated by density eentrifugation and exposed to conditioned medium from rat HCC cell line CSF. Expression of 22 012 genes in quiescent HSC, culture-activated HSC and HCC induction-activated HSC was analyzed by cDNA microarray and confirmed by real-time RT-PCR and Western blot. Results 1672 genes were differentially expressed in culture-activated HSC, including proinflammatory factors, cell adhesion molecules, cell surface receptors, signaling transduction molecules and immune factors. 711 genes were differentially expressed in HCC induction-activated HSC. Some of them were identical to those in culture-activated HSC. HCC Induction-activated HSC showed specific gene expression patterns, including Raft, Rac2, Adam17, Wnt6, MMP-9 and TNF, suggesting that HCC cells can specifically induce HSC activation. Conclusion The gnne expression patterns in HCC induction-activated HSC are different from those in culture-activated HSC. HCC induction-activated HSC may play a major role in the invasion and metastasis of HCC. In vivo activation should be considered as the standard for the study of HSC biology. HCC induction-activated HSC should be considered as the standard for HSC biology studies.