中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2010年
4期
474-477
,共4页
李明岳%余小舫%鲍世韵%林宝行%王春友
李明嶽%餘小舫%鮑世韻%林寶行%王春友
리명악%여소방%포세운%림보행%왕춘우
H3K4m3%H3K9m3%基因表达%胰岛%分化
H3K4m3%H3K9m3%基因錶達%胰島%分化
H3K4m3%H3K9m3%기인표체%이도%분화
Histone 3 lysine 4 trimethylation%Histone 3 lysine 9 trimethylation%Gene expres-sion%Islet%Differentiation
目的 通过比较不同细胞类型之间胰腺十二指肠同源盒1(Pdx-1)、配对盒基因4(Pax4)、MafA(mast cell function associated antigen)和Nkx6.1等胰岛组织特异性基因其转录起始区的H3K4m3和H3K9m3修饰的差异,探讨H3K4m3和H3K9m3修饰对胰岛组织特异性基因表达的作用.方法 采用染色质免疫共沉淀一实时定量聚合酶链反应(PCR)法检测小鼠胚胎干细胞(mES,1×10~7)、小鼠成纤维细胞株NIH3T3细胞(1×10~7)和小鼠β细胞株NIT-1细胞(1×10~7)三者中的胰岛组织特异性基因、Oct4基因和MLH1基因转录起始区H3K4m3和H3K9m3修饰的状况.同时采用实时定量逆转录(RT)-PCR检测上述3种细胞各基因mRNA表达水平.分析H3K4m3和H3K9m3修饰改变与基因表达之间的关系.结果 NIT-1细胞中Pdx-1、Pax4、MafA、Nkx6.1等胰岛组织特异性基因转录起始区的H3K4m的修饰水平分别为:(4.84±0.05)%、(9.91±1.33)%、(10.64±0.87)%、(0.23±0.03)%,与mES细胞比较明显增高(P<0.05),基因表达;NIH3T3细胞中Pdx-1、Pax4、MafA、Nkx6.1等胰岛组织特异性基因转录起始区的H3K9m3的修饰水平分别为:(0.64±0.21)%、(7.04±1.29)%、(0.39±0.10)%、(2.35±0.81)%,与mES细胞比较明显增高(P<0.05),基因不表达.结论 H3K4m3与H3K9m3修饰能相互协调,共同调控胰岛组织特异性基因的表达.
目的 通過比較不同細胞類型之間胰腺十二指腸同源盒1(Pdx-1)、配對盒基因4(Pax4)、MafA(mast cell function associated antigen)和Nkx6.1等胰島組織特異性基因其轉錄起始區的H3K4m3和H3K9m3脩飾的差異,探討H3K4m3和H3K9m3脩飾對胰島組織特異性基因錶達的作用.方法 採用染色質免疫共沉澱一實時定量聚閤酶鏈反應(PCR)法檢測小鼠胚胎榦細胞(mES,1×10~7)、小鼠成纖維細胞株NIH3T3細胞(1×10~7)和小鼠β細胞株NIT-1細胞(1×10~7)三者中的胰島組織特異性基因、Oct4基因和MLH1基因轉錄起始區H3K4m3和H3K9m3脩飾的狀況.同時採用實時定量逆轉錄(RT)-PCR檢測上述3種細胞各基因mRNA錶達水平.分析H3K4m3和H3K9m3脩飾改變與基因錶達之間的關繫.結果 NIT-1細胞中Pdx-1、Pax4、MafA、Nkx6.1等胰島組織特異性基因轉錄起始區的H3K4m的脩飾水平分彆為:(4.84±0.05)%、(9.91±1.33)%、(10.64±0.87)%、(0.23±0.03)%,與mES細胞比較明顯增高(P<0.05),基因錶達;NIH3T3細胞中Pdx-1、Pax4、MafA、Nkx6.1等胰島組織特異性基因轉錄起始區的H3K9m3的脩飾水平分彆為:(0.64±0.21)%、(7.04±1.29)%、(0.39±0.10)%、(2.35±0.81)%,與mES細胞比較明顯增高(P<0.05),基因不錶達.結論 H3K4m3與H3K9m3脩飾能相互協調,共同調控胰島組織特異性基因的錶達.
목적 통과비교불동세포류형지간이선십이지장동원합1(Pdx-1)、배대합기인4(Pax4)、MafA(mast cell function associated antigen)화Nkx6.1등이도조직특이성기인기전록기시구적H3K4m3화H3K9m3수식적차이,탐토H3K4m3화H3K9m3수식대이도조직특이성기인표체적작용.방법 채용염색질면역공침정일실시정량취합매련반응(PCR)법검측소서배태간세포(mES,1×10~7)、소서성섬유세포주NIH3T3세포(1×10~7)화소서β세포주NIT-1세포(1×10~7)삼자중적이도조직특이성기인、Oct4기인화MLH1기인전록기시구H3K4m3화H3K9m3수식적상황.동시채용실시정량역전록(RT)-PCR검측상술3충세포각기인mRNA표체수평.분석H3K4m3화H3K9m3수식개변여기인표체지간적관계.결과 NIT-1세포중Pdx-1、Pax4、MafA、Nkx6.1등이도조직특이성기인전록기시구적H3K4m적수식수평분별위:(4.84±0.05)%、(9.91±1.33)%、(10.64±0.87)%、(0.23±0.03)%,여mES세포비교명현증고(P<0.05),기인표체;NIH3T3세포중Pdx-1、Pax4、MafA、Nkx6.1등이도조직특이성기인전록기시구적H3K9m3적수식수평분별위:(0.64±0.21)%、(7.04±1.29)%、(0.39±0.10)%、(2.35±0.81)%,여mES세포비교명현증고(P<0.05),기인불표체.결론 H3K4m3여H3K9m3수식능상호협조,공동조공이도조직특이성기인적표체.
Objective To explore the regulation of pancreatic islet-specific genes expression by his-tone modification (H3K4m3 and H3Kgm3), through comparing the difference of H3K4m3 and H3K9m3 modifi-cation in the transcription modification site of several pancreatic islet-specific genes (Pdx-1, Pax4, MafA, Nkx6. 1, etc) among different cell types. Methods Chromatin immunoprecipitation-real time quantitative poly-merase chain reaction (PCR) methods were used to study H3K4m3 and H3K9m3 modification in the transcrip-tion initiation site of pancreatic specific-islet genes, Oct4 and MLHI genes, among mouse embryonic stem cells (1×10~7), NIH3T3 cells (1×10~7) and NIT-1 cells (1×10~7). At the same time, the mRNA levels of these genes were quantified among the above three cell types using reverse transcription (RT)-PCR. The relationship between the differences in H3K4m3 and H3K9m3 modification and gene expression was analyzed. Results (1) In NIT-1 cells, the levels of H3K4m3 modification in the transcription-initiation-site of Pdx-1, Pax4, MafA, Nkx6. 1 were (4.84±0.05)%, (9.91±1.33)%, (10.64±0.87)%, and (0.23±0.03)% respectively. Com-pared with ruES cells, NIT-1 cells had a sigificanfly higher level of H3K4m modification (P <0. 05), with a si-gificant level of gene expression; (2) In NIH3T3 cells, the levels of H3K9m3 modification in the transcription-initiation-site of Pdx-1, Pax4, MafA, Nkx6. 1 were (0. 64±0. 21 )%, (7.04 ± 1.29)%, (0. 39±0. 10)%, (2.35±0. 81)% respectively. Compared with ruES cells, NIH3T3 cell had a sigifieantly higher level of H3K9m3 modification (P<0.05), with no gene expression. Conclusion H3K4m3 and H3K9m3 modification can regulate each other, and synergtically regulate the expression of pancreatic islet-specific genes.
