中华放射肿瘤学杂志
中華放射腫瘤學雜誌
중화방사종류학잡지
CHINESE JOURNAL OF RADIATION ONCOLOGY
2008年
6期
467-469
,共3页
廖遇平%丁思娟%周蓉蓉%肖华平
廖遇平%丁思娟%週蓉蓉%肖華平
료우평%정사연%주용용%초화평
细胞系,肿瘤%放射敏感性%E1A基因%血管内皮生长因子
細胞繫,腫瘤%放射敏感性%E1A基因%血管內皮生長因子
세포계,종류%방사민감성%E1A기인%혈관내피생장인자
Cell line,tumor%Radiosensitivity%EIA gene%Vascular endothelial growth factor
目的 探讨ElA基因对喉癌细胞系(Hep-2细胞)放射敏感性的影响并初步探讨其机制.方法 以腺病毒为载体将E1A基凶及其空载体转染至人Hep-2细胞,经RT-PCR法鉴定获得含ElA的阳性兜隆.设未转染组(PBS)、转染空载体组(Ad-β-gal)和转染组(Ad-E1A),分别给予6 MVx线单次照射0、1、2、4、6、8、10 Gy,成克降实验绘制细胞存活曲线并计算D0、Dq、α、β值以观察E1A基因对Hep-2细胞放射敏感件的影响.设PBS组、Ad-β-gal组、Ad-E1A组、PBS+照射组、Ad-β-gal+照射组、Ad-E1A+照射组,单次照射6 Gy,流式细胞仪检测细胞凋亡及RT-PCR检测血管内皮牛长因子(VEGF)水平并半定量VEGF以初步探讨机制.结果 转染后的阳性克隆细胞RT-PCR结果显示E1A已整合到细胞基因组中并且稳定表达.Ad-E1A组、Ad-β-gal组、PBS组细胞的D0值分别为0.86、1.96、1.98 Gy,Dq值分别为1.02、1.97、1.99 Gy,α值分别为0.536、0.112、0.104(Gy-1)和β值分别为0.521、0.137、0.125(Gy-2).PBS组、Ad-β-gal组、Ad-ElA组、PBS+照射组、Ad-β-gal+照射组、Ad-E1A+照射组的细胞凋亡率分别为1.26%、1.18%、2.16%、2.55%、2.96%、4.96%.Ad-E1A组、PBS+照射组、Ad-βgal+照射组及Ad-ElA+照射组VEGF的表达水平较PBS组及Ad-β-gal组低,且Ad-E1A+照射组表达最低.结论 成功构建了稳定表达E1A基因的喉癌细胞系.EIA基因对人喉癌细胞有放射增敏作用,其机制可能与降低VEGF表达和促进细胞凋亡有关.
目的 探討ElA基因對喉癌細胞繫(Hep-2細胞)放射敏感性的影響併初步探討其機製.方法 以腺病毒為載體將E1A基兇及其空載體轉染至人Hep-2細胞,經RT-PCR法鑒定穫得含ElA的暘性兜隆.設未轉染組(PBS)、轉染空載體組(Ad-β-gal)和轉染組(Ad-E1A),分彆給予6 MVx線單次照射0、1、2、4、6、8、10 Gy,成剋降實驗繪製細胞存活麯線併計算D0、Dq、α、β值以觀察E1A基因對Hep-2細胞放射敏感件的影響.設PBS組、Ad-β-gal組、Ad-E1A組、PBS+照射組、Ad-β-gal+照射組、Ad-E1A+照射組,單次照射6 Gy,流式細胞儀檢測細胞凋亡及RT-PCR檢測血管內皮牛長因子(VEGF)水平併半定量VEGF以初步探討機製.結果 轉染後的暘性剋隆細胞RT-PCR結果顯示E1A已整閤到細胞基因組中併且穩定錶達.Ad-E1A組、Ad-β-gal組、PBS組細胞的D0值分彆為0.86、1.96、1.98 Gy,Dq值分彆為1.02、1.97、1.99 Gy,α值分彆為0.536、0.112、0.104(Gy-1)和β值分彆為0.521、0.137、0.125(Gy-2).PBS組、Ad-β-gal組、Ad-ElA組、PBS+照射組、Ad-β-gal+照射組、Ad-E1A+照射組的細胞凋亡率分彆為1.26%、1.18%、2.16%、2.55%、2.96%、4.96%.Ad-E1A組、PBS+照射組、Ad-βgal+照射組及Ad-ElA+照射組VEGF的錶達水平較PBS組及Ad-β-gal組低,且Ad-E1A+照射組錶達最低.結論 成功構建瞭穩定錶達E1A基因的喉癌細胞繫.EIA基因對人喉癌細胞有放射增敏作用,其機製可能與降低VEGF錶達和促進細胞凋亡有關.
목적 탐토ElA기인대후암세포계(Hep-2세포)방사민감성적영향병초보탐토기궤제.방법 이선병독위재체장E1A기흉급기공재체전염지인Hep-2세포,경RT-PCR법감정획득함ElA적양성두륭.설미전염조(PBS)、전염공재체조(Ad-β-gal)화전염조(Ad-E1A),분별급여6 MVx선단차조사0、1、2、4、6、8、10 Gy,성극강실험회제세포존활곡선병계산D0、Dq、α、β치이관찰E1A기인대Hep-2세포방사민감건적영향.설PBS조、Ad-β-gal조、Ad-E1A조、PBS+조사조、Ad-β-gal+조사조、Ad-E1A+조사조,단차조사6 Gy,류식세포의검측세포조망급RT-PCR검측혈관내피우장인자(VEGF)수평병반정량VEGF이초보탐토궤제.결과 전염후적양성극륭세포RT-PCR결과현시E1A이정합도세포기인조중병차은정표체.Ad-E1A조、Ad-β-gal조、PBS조세포적D0치분별위0.86、1.96、1.98 Gy,Dq치분별위1.02、1.97、1.99 Gy,α치분별위0.536、0.112、0.104(Gy-1)화β치분별위0.521、0.137、0.125(Gy-2).PBS조、Ad-β-gal조、Ad-ElA조、PBS+조사조、Ad-β-gal+조사조、Ad-E1A+조사조적세포조망솔분별위1.26%、1.18%、2.16%、2.55%、2.96%、4.96%.Ad-E1A조、PBS+조사조、Ad-βgal+조사조급Ad-ElA+조사조VEGF적표체수평교PBS조급Ad-β-gal조저,차Ad-E1A+조사조표체최저.결론 성공구건료은정표체E1A기인적후암세포계.EIA기인대인후암세포유방사증민작용,기궤제가능여강저VEGF표체화촉진세포조망유관.
Objective To investigate the effect of El A gene on the radiosensitivity of human laryngeal carcinoma cells and its correlated mechanisms. Methods The Ad-E1A and Ad-β-gal were amplifieated in Hek293 cells, extracted by freezing (-80℃) and thawing(37℃) repeatedly (3 times) , purificated by the method of density gradient of CsC1 and titrated by plaque assay method. Then they were transfected into human laryngeal carcinoma cells (Hep-2) and authenticated by RT-PCR. The radiosensitivity of Hep-2 cells transfeeted with or without El A were studied by cell surviral curve. Finally we investigated the correlated mechanisms including cell apoptosis studied by flow cytometry and VEGF content studied by RT-PCR. Resuits The radiosensitivity of Hep-2 cells transfected with El A was intensified, Do and Dq were lowered and α was increased. Flow cytometry showed that the apoptosis rate of cells with E1A or with El A and radiotherapy was increased. The VEGF content of the cells transfeeted with E1 A or treated by radiotherapy was decreased, which reached the lowest level when the cells were treated with the both mathods. Conclusions E1 A gene can intensify the radiosensitivity and contribute to the apoptosis of human laryngeal carcinoma cells. El A gene and radiotherapy can markedly decrease the VEGF content.
