贵州医药
貴州醫藥
귀주의약
GUIZHOU MEDICAL JOURNAL
2014年
9期
771-775
,共5页
范芳%安国顺%李淑艳%倪菊华%贾弘禔
範芳%安國順%李淑豔%倪菊華%賈弘禔
범방%안국순%리숙염%예국화%가홍제
八氯腺苷%细胞周期%DNA依赖蛋白激酶催化亚基%拓扑异构酶I
八氯腺苷%細胞週期%DNA依賴蛋白激酶催化亞基%拓撲異構酶I
팔록선감%세포주기%DNA의뢰단백격매최화아기%탁복이구매I
8-chloro-adenosine%Cell cycle%DNA-PKcs%T o po Ⅰ
目的：八氯腺苷对肺癌细胞株A549和H1299细胞增殖及细胞周期的影响及其机制探讨。方法 A549和 H1299经10．0μmol/L 八氯腺苷处理后，MTT法检测细胞生长和增殖情况，流式细胞术检测细胞周期，Western blot检测DNA-PKcs、γ-H2AX、Ku80、Ku70及 Topo Ⅰ的表达情况。结果 MTT法检测结果显示，用10．0μmol/L八氯腺苷分别处理 A549、H1299细胞24、48、72 h后，A549、H1299细胞生长被抑制；流式细胞分析则显示，A549、H1299细胞G2/M 期细胞数增多；Western blot结果表明，γ-H2AX的表达均增加，DNA-PKcs蛋白的表达均下降，而Ku80、Ku70的表达未见明显变化；经八氯腺苷处理后Topo Ⅰ蛋白在A549细胞中的表达逐渐增加，而在 H1299细胞中的表达却逐渐下降。结论八氯腺苷可抑制A549和 H1299细胞生长，诱导细胞发生G2/M 期阻滞，其机制可能与DNA-PKcs表达下调有关。在A549细胞中Topo Ⅰ可能通过p53依赖途径介导DNA损伤修复，从而引发对细胞的毒性作用；而在p53缺失的H1299细胞中可能是通过其它途径调控细胞周期。
目的：八氯腺苷對肺癌細胞株A549和H1299細胞增殖及細胞週期的影響及其機製探討。方法 A549和 H1299經10．0μmol/L 八氯腺苷處理後，MTT法檢測細胞生長和增殖情況，流式細胞術檢測細胞週期，Western blot檢測DNA-PKcs、γ-H2AX、Ku80、Ku70及 Topo Ⅰ的錶達情況。結果 MTT法檢測結果顯示，用10．0μmol/L八氯腺苷分彆處理 A549、H1299細胞24、48、72 h後，A549、H1299細胞生長被抑製；流式細胞分析則顯示，A549、H1299細胞G2/M 期細胞數增多；Western blot結果錶明，γ-H2AX的錶達均增加，DNA-PKcs蛋白的錶達均下降，而Ku80、Ku70的錶達未見明顯變化；經八氯腺苷處理後Topo Ⅰ蛋白在A549細胞中的錶達逐漸增加，而在 H1299細胞中的錶達卻逐漸下降。結論八氯腺苷可抑製A549和 H1299細胞生長，誘導細胞髮生G2/M 期阻滯，其機製可能與DNA-PKcs錶達下調有關。在A549細胞中Topo Ⅰ可能通過p53依賴途徑介導DNA損傷脩複，從而引髮對細胞的毒性作用；而在p53缺失的H1299細胞中可能是通過其它途徑調控細胞週期。
목적：팔록선감대폐암세포주A549화H1299세포증식급세포주기적영향급기궤제탐토。방법 A549화 H1299경10．0μmol/L 팔록선감처리후，MTT법검측세포생장화증식정황，류식세포술검측세포주기，Western blot검측DNA-PKcs、γ-H2AX、Ku80、Ku70급 Topo Ⅰ적표체정황。결과 MTT법검측결과현시，용10．0μmol/L팔록선감분별처리 A549、H1299세포24、48、72 h후，A549、H1299세포생장피억제；류식세포분석칙현시，A549、H1299세포G2/M 기세포수증다；Western blot결과표명，γ-H2AX적표체균증가，DNA-PKcs단백적표체균하강，이Ku80、Ku70적표체미견명현변화；경팔록선감처리후Topo Ⅰ단백재A549세포중적표체축점증가，이재 H1299세포중적표체각축점하강。결론팔록선감가억제A549화 H1299세포생장，유도세포발생G2/M 기조체，기궤제가능여DNA-PKcs표체하조유관。재A549세포중Topo Ⅰ가능통과p53의뢰도경개도DNA손상수복，종이인발대세포적독성작용；이재p53결실적H1299세포중가능시통과기타도경조공세포주기。
Objective To explore the mechanisms of 8-chloro-adenosine-induced G2/M arrest in A549 and H1299 Cells .Methods The cell growth and cell cycle of A549 and H1299 were detected by MTT and flow cytometry analysis after 8-Cl-Ado exposures .The protein expression of DNA-PKcs ,γ-H2AX ,Ku80 ,Ku70 and Topo Ⅰ were detected by western blot (WB) assay .Results The MTT re-sults showed that the cell growth of A549 and H1299 cells were inhibited after 8-Cl-Ado exposures for 24 ,48 ,72 h .Flow cytometry analysis showed that the cells in G2/M phase of A549 and H1299 cells was increased .WB showed that the levels of γ-H2AX was upregulated ,DNA-PKcs was downregulat-ed in both A549 and H1299 cells ,meanwhile ,Ku80 and Ku70 expressions in the two cells did not see obvious change .The expression of Topo Ⅰ gradually increased in A549 cells ,and gradually decline in H1299 cells after 8-Cl-Ado treatment .Conclusion 8-Cl-Ado can inhibit the growth of A549 and H1299 cells ,induce cells in G2 /M phase retardation ,its mechanism may be related to DNA-PKcs expres-sion .Topo I in A549 cells may be mediated by p53 dependent way DNA damage repair ,causing toxic effect on cells .In loss of p53 H1299 cells may be initiate cell cycle arrest through other channels .
