中华内分泌代谢杂志
中華內分泌代謝雜誌
중화내분비대사잡지
CHINESE JOURNAL OF ENDOCRINOLOGY AND METABOLISM
2009年
5期
544-548
,共5页
Ku70%反义寡核苷酸%甲状腺肿瘤%细胞凋亡%辐射敏感性
Ku70%反義寡覈苷痠%甲狀腺腫瘤%細胞凋亡%輻射敏感性
Ku70%반의과핵감산%갑상선종류%세포조망%복사민감성
Ku70%Oligonucleotides，antisense%Thyroid neoplasms%Apoptosis%Radiosensitivity
目的 研究DNA修复基因Ku70反义寡核苷酸(ASODN)对甲状腺癌细胞辐射敏感性的影响.方法 设计合成特异性靶向Ku70的ASODN,以不同浓度转染人甲状腺髓样癌细胞(TT),并设脂质体和正义寡核苷酸(SODN)对照组进行比较.采用Western印迹技术检测各组细胞中Ku70蛋白表达水平.利用不同剂量60 Co γ射线照射细胞后,细胞克隆形成实验检测细胞存活分数.CCK-8法、Annexin-V/PI染色法分析ASODN对细胞活力和细胞凋亡的影响.彗星电泳法比较γ射线照射后DNA双链断裂的修复效率.结果 转染ASODN各组细胞Ku70蛋白表达均较脂质体对照组、SODN组明显降低,并呈浓度依赖性;照射后细胞存活率降低(P＜0.01),凋亡率显著增加(P＜0.01),DNA双链断裂的修复效率降低(P＜0.01).结论 ASODN能够特异性地下调甲状腺癌细胞Ku70的表达,抑制γ射线照射后细胞存活和DNA双链断裂修复,提高肿瘤细胞的辐射敏感性.
目的 研究DNA脩複基因Ku70反義寡覈苷痠(ASODN)對甲狀腺癌細胞輻射敏感性的影響.方法 設計閤成特異性靶嚮Ku70的ASODN,以不同濃度轉染人甲狀腺髓樣癌細胞(TT),併設脂質體和正義寡覈苷痠(SODN)對照組進行比較.採用Western印跡技術檢測各組細胞中Ku70蛋白錶達水平.利用不同劑量60 Co γ射線照射細胞後,細胞剋隆形成實驗檢測細胞存活分數.CCK-8法、Annexin-V/PI染色法分析ASODN對細胞活力和細胞凋亡的影響.彗星電泳法比較γ射線照射後DNA雙鏈斷裂的脩複效率.結果 轉染ASODN各組細胞Ku70蛋白錶達均較脂質體對照組、SODN組明顯降低,併呈濃度依賴性;照射後細胞存活率降低(P＜0.01),凋亡率顯著增加(P＜0.01),DNA雙鏈斷裂的脩複效率降低(P＜0.01).結論 ASODN能夠特異性地下調甲狀腺癌細胞Ku70的錶達,抑製γ射線照射後細胞存活和DNA雙鏈斷裂脩複,提高腫瘤細胞的輻射敏感性.
목적 연구DNA수복기인Ku70반의과핵감산(ASODN)대갑상선암세포복사민감성적영향.방법 설계합성특이성파향Ku70적ASODN,이불동농도전염인갑상선수양암세포(TT),병설지질체화정의과핵감산(SODN)대조조진행비교.채용Western인적기술검측각조세포중Ku70단백표체수평.이용불동제량60 Co γ사선조사세포후,세포극륭형성실험검측세포존활분수.CCK-8법、Annexin-V/PI염색법분석ASODN대세포활력화세포조망적영향.혜성전영법비교γ사선조사후DNA쌍련단렬적수복효솔.결과 전염ASODN각조세포Ku70단백표체균교지질체대조조、SODN조명현강저,병정농도의뢰성;조사후세포존활솔강저(P＜0.01),조망솔현저증가(P＜0.01),DNA쌍련단렬적수복효솔강저(P＜0.01).결론 ASODN능구특이성지하조갑상선암세포Ku70적표체,억제γ사선조사후세포존활화DNA쌍련단렬수복,제고종류세포적복사민감성.
Objective To investigate the effect of Ku70 antisense oligonueleotide(ASODN) on the radiosensitivity of thyroid carcinoma cells.Methods Targeted ASODN of Ku70 was synthesized and transfected into human medullary thyroid carcinoma cell lines at different concentrations,with lipofectamine and sense oligonucleotides (SODN) as control groups.Western blotting was used to detect the expression level of Ku70 protein.Cell clonogenic assay was performed to determine the cell survival fraction after 60 Co γ ray radiation.CCK-8 assay and Annexin-V/PI staining were applied to evaluate the cell viability and apoptosis after radiation.The DNA repair efficiency for double-strand breaks (DSB) was evaluated by Comet assay.Results Compared with lipofectamine and SODN control groups,the expression of Ku70 protein in cells of ASODN group was obviously decreased.The cell survival was markedly inhibited by ASODN in a dose dependent manner and apoptosis rate significantly increased (P ＜0.01).The repair efficiency for radiation-induced DNA DSBs was makedly reduced by ASODN (P ＜0.01).Conlusion ASODN specifically down-regulates the expression of KuT0 in thyroid carcinoma cells.The cell radiosensitivity was greatly enhanced as evidenced by inhibition of cell survival,increase in apoptosis and suppression of DNA DSBs repair.
