遗传学报
遺傳學報
유전학보
ACTA GENETICA SINICA
2006年
10期
881-891
,共11页
王水良%兰风华%庄岳鹏%李慧忠%黄梁浒%郑德柱%曾健%董荔红%朱忠勇%傅继梁
王水良%蘭風華%莊嶽鵬%李慧忠%黃樑滸%鄭德柱%曾健%董荔紅%硃忠勇%傅繼樑
왕수량%란풍화%장악붕%리혜충%황량호%정덕주%증건%동려홍%주충용%부계량
基因表达谱%hLRH-1%RNA干扰%表达稳定抑制
基因錶達譜%hLRH-1%RNA榦擾%錶達穩定抑製
기인표체보%hLRH-1%RNA간우%표체은정억제
gene expression profile%hLRH-1%RNA interference%stable suppression
为建立一hLRH-1表达稳定抑制的细胞系,我们构建了hLRH-J基因靶向的稳定RNA干涉载体(pSineohLRH-1)并导入肝细胞癌细胞BEL-7402.通过半定量RT-PCR分析发现,携有pSineohLRH-1的BEL-7402细胞其hLRH-1基因mRNA的表达抑制率达60%.此外,微阵列法基因表达谱分析结果表明,与未干涉的对照细胞相比,包括一些肿瘤相关的基因如Gadd45β和PTEN在内的405个基因在hLRH-1基因表达稳定下调的BEL-7402细胞中呈明显的表达差异,意示hLRH-1具比已知更为广泛的生物学功能.尽管hLRH-1与这些差异表达基因的确切关系尚有待进一步的实验探究,我们的发现仍为hLRH-1在肿瘤发生发展中可能的作用机制的揭示提供了新的线索.
為建立一hLRH-1錶達穩定抑製的細胞繫,我們構建瞭hLRH-J基因靶嚮的穩定RNA榦涉載體(pSineohLRH-1)併導入肝細胞癌細胞BEL-7402.通過半定量RT-PCR分析髮現,攜有pSineohLRH-1的BEL-7402細胞其hLRH-1基因mRNA的錶達抑製率達60%.此外,微陣列法基因錶達譜分析結果錶明,與未榦涉的對照細胞相比,包括一些腫瘤相關的基因如Gadd45β和PTEN在內的405箇基因在hLRH-1基因錶達穩定下調的BEL-7402細胞中呈明顯的錶達差異,意示hLRH-1具比已知更為廣汎的生物學功能.儘管hLRH-1與這些差異錶達基因的確切關繫尚有待進一步的實驗探究,我們的髮現仍為hLRH-1在腫瘤髮生髮展中可能的作用機製的揭示提供瞭新的線索.
위건립일hLRH-1표체은정억제적세포계,아문구건료hLRH-J기인파향적은정RNA간섭재체(pSineohLRH-1)병도입간세포암세포BEL-7402.통과반정량RT-PCR분석발현,휴유pSineohLRH-1적BEL-7402세포기hLRH-1기인mRNA적표체억제솔체60%.차외,미진렬법기인표체보분석결과표명,여미간섭적대조세포상비,포괄일사종류상관적기인여Gadd45β화PTEN재내적405개기인재hLRH-1기인표체은정하조적BEL-7402세포중정명현적표체차이,의시hLRH-1구비이지경위엄범적생물학공능.진관hLRH-1여저사차이표체기인적학절관계상유대진일보적실험탐구,아문적발현잉위hLRH-1재종류발생발전중가능적작용궤제적게시제공료신적선색.
To establish a cell line with a permanent suppression of hLRH-1 in this study, a stable RNAi vector (pSineohLRH-1)targeting hLRH-1 was constructed and introduced into hepatocellular carcinoma cell, BEL-7402. By semiquantitative RT-PCR analysis, the expression of hLRH-1 in BEL-7402 cells carrying pSineohLRH-1 was shown to be significantly suppressed by up to ～60%. In addition, microarray analysis was carried out to assess the extent of altered gene expression in BEL-7402 cells with stable knockdown of hLRH-1. Direct comparison of gene-expression profiles of more than 18 000 genes showed that 405 of the expressed genes in hLRH-1-knockdown cells differed dramatically in expression levels from those in controls, which suggested the even extensive biological functions of hLRH-1. Interestingly, among those differentially expressed genes, some are cancer-associated such as Gadd45β and PTEN, and their expressions were further validated. Although the identification of the exact relationship between these genes and hLRH-1 awaits intensive investigation, the findings of this study provide new insights into the mechanism by which hLRH-1 is involved in tumorigenesis.
