CAJ | 학술논문

目的观察组蛋白H3K9me3修饰在胆管癌发病机制中的作用.方法收集9例病理确诊的胆管腺癌组织和5例正常的肝外胆管组织,采用染色质免疫共沉淀联合芯片技术(ChIP-chip)检测两种组织细胞全基因组的蛋白H3K9me3修饰水平,进行基因验证,并检测mRNA的表达水平.结果与胆管组织细胞比较,胆管癌细胞全基因组范围共筛选出67个基因存在H3K9me3修饰显著差异,其中35个基因升高,32个基因降低.在癌细胞中,6个挑选基因[钙结合蛋白A2(S100A2)、GTP酶激活蛋白基因2亚型(ANAPC2)、CBFA2T3、GTP酶激活蛋白4(ARHGAP)、鸟嘌呤核苷酸交换因子3(RAPGEF3)、α型蛋白激酶C基因(PRKCA)]的H3K9me3修饰水平分别为:(3.51±0.21)％、(8.45±0.16)％、(0.89±0.27)％、(0.66±0.22)％、(0.04±0.01)％、(5.21±1.53)％,与正常胆管细胞比较差异有统计学意义,并且其修饰水平与基因表达呈负直线相关(r=-0.539,P＜0.05).结论与正常胆管细胞比较,胆管癌细胞中多个基因的组蛋白H3 Kme3修饰水平存在显著改变;并导致了多个相关的癌基因及抑癌基因转录表达的改变.
목적 관찰조단백H3K9me3수식재담관암발병궤제중적작용.방법 수집9례병리학진적담관선암조직화5례정상적간외담관조직,채용염색질면역공침정연합심편기술(ChIP-chip)검측량충조직세포전기인조적단백H3K9me3수식수평,진행기인험증,병검측mRNA적표체수평.결과 여담관조직세포비교,담관암세포전기인조범위공사선출67개기인존재H3K9me3수식현저차이,기중35개기인승고,32개기인강저.재암세포중,6개도선기인[개결합단백A2(S100A2)、GTP매격활단백기인2아형(ANAPC2)、CBFA2T3、GTP매격활단백4(ARHGAP)、조표령핵감산교환인자3(RAPGEF3)、α형단백격매C기인(PRKCA)]적H3K9me3수식수평분별위:(3.51±0.21)％、(8.45±0.16)％、(0.89±0.27)％、(0.66±0.22)％、(0.04±0.01)％、(5.21±1.53)％,여정상담관세포비교차이유통계학의의,병차기수식수평여기인표체정부직선상관(r=-0.539,P＜0.05).결론 여정상담관세포비교,담관암세포중다개기인적조단백H3 Kme3수식수평존재현저개변;병도치료다개상관적암기인급억암기인전록표체적개변.
Objective To study the effect of histone H3 lysine 9 trimethylation (H3K9me3) modification on cholangiocarcinoma moleculor mechanism.Methods 9 cases cholangiocarcinoma tissue and 5 cases normal extrahepatic bile duct tissue were collected.Chromatin immunoprecipitation linked to microarrays (ChIP-chip) was adopted to profile the variations of histone H3K9me3 in genome-wide in cholangiocarcinoma cell and normal extrahepatic bile duct cell.CHIP-quantitative polymerase chain reaction (qPCR) was used to validate the microrray results.mRNA expression analysis of 6 selected genes by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was performed to confirm the correlations between H3K9me3 and gene expression.Results 67 (35 increased and 32 decreased) gene displayed significant histone H3K9me3 differences were found in tumor cell compared with normal extrahepatic bile duct cell.In bile duct cancer tissues,six selected oncogenes [calcium binding protein A2 (S100A2),anaphase-promoting complex subunit 2 (ANAPC2),myeloid translocation gene on chromosome 16 (CBFA2T3),Rho GTPase activating protein 4 isoform 2 (ARHGAP),Rap guanine nucleotide exchange factor 3 isoform (RAPGEF3),protein kinase C alpha (PRKCA)] H3K9me3 modifications levelwere:(3.51±3.51) ％,(8.45±0.16) ％,(0.89 ±0.27) ％,(0.66 ±0.22) ％,(0.04± 0.01) ％,(5.21 ± 1.53) ％,there was a statistically significant difference compared with normal bile duct cells.It was the Pearson correlation between selected genes expression and H3K9Me3 modification (r =-0.539) in the turmor cell.Conclusion Compared with normal extrahepatic bile duct cell,there are significant changes in many genes of cholangiocarcinoma cell about histone H3K9Me3 profiling.And resulting in a number of related oncogenes and tumor suppressor genes expression change.