中华医学杂志(英文版)
中華醫學雜誌(英文版)
중화의학잡지(영문판)
CHINESE MEDICAL JOURNAL
2001年
12期
1295-1299
,共5页
邓宇斌%李树浓%李夏新%Teyssier Magali
鄧宇斌%李樹濃%李夏新%Teyssier Magali
산우빈%리수농%리하신%Teyssier Magali
辐射%电离%范可尼贫血%基因表达%hHR21sp基因
輻射%電離%範可尼貧血%基因錶達%hHR21sp基因
복사%전리%범가니빈혈%기인표체%hHR21sp기인
ionizing radiation%Fanconi's anemia%gene expression%hHR21sp%gene
目的检测UV和γ辐射对正常人外周血单核细胞的hHR21sp基因转录表达水平及hHR21sp在范可尼贫血（Fanconis Anemia FA）骨髓造血细胞和激活后的外周血单核细胞的转录表达情况,并分析hHR21sp基因在FA贫血发病机制中的作用。方法对范可尼贫血骨髓造血细胞和激活后的外周血单核细胞和正常人外周血单核细胞在UV或γ辐射后不同时间提取细胞总RNA，通过RT-PCR、Southern杂交、以β-actin为内参照放射影像对hHR21sp基因的转录表达水平进行检测,比较FA骨髓造血细胞与对照组差异和激活前后FA外周血单核细胞hHR21sp基因的表达水平。结果正常人外周血单核细胞与对照组比较在80j/m2 UV辐射hHR21sp表达于3h开始增加；在6h表达水平最高，与正常对照有2倍差异（P<0.05）；而γ-辐射5 Gy辐射6 h增加明显，在9h后有所降低与正常对照有2倍多差异（P<0.01）。可见FA病人骨髓造血细胞和对照组的hHR21sp基因表达水平有明显差异（P<0.05）。实验发现在激活后对FA病人外周血单核细胞检测，hHR21sp基因表达较对照组PB-MNC降低达2倍之多（P<0.05）。结论 hHR21sp表达水平在一定剂量电离辐射范围内随辐射剂量增加而增高，且对γ-辐射较UV辐射敏感，FA病人骨髓造血细胞和对照组的hHR21sp基因表达有明显差异，激活后的FA病人外周血单核细胞 hHR21sp基因表达降低显著，提示FA病人造血细胞hHR21sp基因表达缺陷，本研究结果为下一步深入研究FA发病机制提供实验基础。
目的檢測UV和γ輻射對正常人外週血單覈細胞的hHR21sp基因轉錄錶達水平及hHR21sp在範可尼貧血（Fanconis Anemia FA）骨髓造血細胞和激活後的外週血單覈細胞的轉錄錶達情況,併分析hHR21sp基因在FA貧血髮病機製中的作用。方法對範可尼貧血骨髓造血細胞和激活後的外週血單覈細胞和正常人外週血單覈細胞在UV或γ輻射後不同時間提取細胞總RNA，通過RT-PCR、Southern雜交、以β-actin為內參照放射影像對hHR21sp基因的轉錄錶達水平進行檢測,比較FA骨髓造血細胞與對照組差異和激活前後FA外週血單覈細胞hHR21sp基因的錶達水平。結果正常人外週血單覈細胞與對照組比較在80j/m2 UV輻射hHR21sp錶達于3h開始增加；在6h錶達水平最高，與正常對照有2倍差異（P<0.05）；而γ-輻射5 Gy輻射6 h增加明顯，在9h後有所降低與正常對照有2倍多差異（P<0.01）。可見FA病人骨髓造血細胞和對照組的hHR21sp基因錶達水平有明顯差異（P<0.05）。實驗髮現在激活後對FA病人外週血單覈細胞檢測，hHR21sp基因錶達較對照組PB-MNC降低達2倍之多（P<0.05）。結論 hHR21sp錶達水平在一定劑量電離輻射範圍內隨輻射劑量增加而增高，且對γ-輻射較UV輻射敏感，FA病人骨髓造血細胞和對照組的hHR21sp基因錶達有明顯差異，激活後的FA病人外週血單覈細胞 hHR21sp基因錶達降低顯著，提示FA病人造血細胞hHR21sp基因錶達缺陷，本研究結果為下一步深入研究FA髮病機製提供實驗基礎。
목적검측UV화γ복사대정상인외주혈단핵세포적hHR21sp기인전록표체수평급hHR21sp재범가니빈혈（Fanconis Anemia FA）골수조혈세포화격활후적외주혈단핵세포적전록표체정황,병분석hHR21sp기인재FA빈혈발병궤제중적작용。방법대범가니빈혈골수조혈세포화격활후적외주혈단핵세포화정상인외주혈단핵세포재UV혹γ복사후불동시간제취세포총RNA，통과RT-PCR、Southern잡교、이β-actin위내삼조방사영상대hHR21sp기인적전록표체수평진행검측,비교FA골수조혈세포여대조조차이화격활전후FA외주혈단핵세포hHR21sp기인적표체수평。결과정상인외주혈단핵세포여대조조비교재80j/m2 UV복사hHR21sp표체우3h개시증가；재6h표체수평최고，여정상대조유2배차이（P<0.05）；이γ-복사5 Gy복사6 h증가명현，재9h후유소강저여정상대조유2배다차이（P<0.01）。가견FA병인골수조혈세포화대조조적hHR21sp기인표체수평유명현차이（P<0.05）。실험발현재격활후대FA병인외주혈단핵세포검측，hHR21sp기인표체교대조조PB-MNC강저체2배지다（P<0.05）。결론 hHR21sp표체수평재일정제량전리복사범위내수복사제량증가이증고，차대γ-복사교UV복사민감，FA병인골수조혈세포화대조조적hHR21sp기인표체유명현차이，격활후적FA병인외주혈단핵세포 hHR21sp기인표체강저현저，제시FA병인조혈세포hHR21sp기인표체결함，본연구결과위하일보심입연구FA발병궤제제공실험기출。
Objective The radiation sensitive gene rad 21 of Schizosaccharomyces pombe is involved in the repair of double-stranded breaks in DNA and is essential for mitotic growth. The hHR21sp gene is its human homologue. In an attempt to investigate the role of hHR21sp in DNA repair, we studied the effects of UV and γ-ray irradiation on hHR21sp gene expression in normal human peripheral blood cells, and non-iradiated peripheral and bone marrow cells from Fanconi anemia (FA) patients who have shown DNA repair deficiency.Methods Total steady state RNA was extracted from peripheral blood cells and bone marrow. RNA transcripts were quantified after RT-PCR and Southern blot, phosphoimmage and autoradiogram analysis. The results were compared with control groups. Results hHR21sp expression was significantly increased from 3h to 9h after UV irradiation in peripheral blood cells from normal subjects at doses of 40-80j/m2 (P<0.05). hHR21sp was also up-regulated by γ-ray irradiation at 6h to 9h at dose of 1 to 5Gy (P<0.01), which was more significant than the UV irradiation. In the non-irradiated FA patient group, hHR21sp expression was decreased in bone marrow hematopoietic cells (P<0.05). After activation by PHA and IL-2, there was still a significant depression in expression by the FA patients peripheral blood cells compared with control groups (P<0.05). Conclusion hHR21sp was up-regulated at doses and times irradiated at the range tested in normal peripheral blood cells, and is more affected by γ-ray irradiation than UV irradiation. FA patient bone marrow hematopoietic cells and peripheral blood mononuclear cells showed down-regulation of hHR21sp expression. The results imply that defects in DNA repair via hHR21sp expression may play an important role in the pathogenesis of FA syndrome.
