CAJ | 학술논문

目的：研究重离子对人外周血T淋巴细胞增殖、凋亡等生物学性能的影响并探讨其机制，为肿瘤放射治疗的辐射防护提供实验依据和基础。方法 Ficoll分离法分离人外周血T淋巴细胞，采用12 C重离子束坪区照射，照射样品能量为70 MeV、LET＝29 keV/μm，照射剂量为1．0和2．0 Gy，剂量率为0．5 Gy/min。分别于照射后12、24 h，RT-PCR检测凋亡相关基因Bcl-2， Bax， Caspase3， Caspase8和Caspase9的表达；于照射后24、48 h，CCK8法检测细胞增殖能力；于照射后24、48 h，采用AnnexinV-PE/7-AAD、AnnexinV-FITC/PI法检测凋亡发生，并采用RT-PCR检测凋亡相关蛋白Bcl-2、Bax和Caspase3的表达。结果重离子照射可明显抑制人外周血T淋巴细胞的增殖，随着剂量增大，抑制作用更加明显。同时，重离子照射可促进T淋巴细胞的凋亡，特别是对于晚期凋亡的诱导作用（P＜0．01）。 RT-PCR检测结果显示，重离子辐射可抑制抗凋亡蛋白Bcl-2的表达，促进促凋亡蛋白Bax和Caspase3的表达（P＜0．01）。结论重离子辐射可显著影响抑制T淋巴细胞的增殖并促进其凋亡。
목적：연구중리자대인외주혈T림파세포증식、조망등생물학성능적영향병탐토기궤제，위종류방사치료적복사방호제공실험의거화기출。방법 Ficoll분리법분리인외주혈T림파세포，채용12 C중리자속평구조사，조사양품능량위70 MeV、LET＝29 keV/μm，조사제량위1．0화2．0 Gy，제량솔위0．5 Gy/min。분별우조사후12、24 h，RT-PCR검측조망상관기인Bcl-2， Bax， Caspase3， Caspase8화Caspase9적표체；우조사후24、48 h，CCK8법검측세포증식능력；우조사후24、48 h，채용AnnexinV-PE/7-AAD、AnnexinV-FITC/PI법검측조망발생，병채용RT-PCR검측조망상관단백Bcl-2、Bax화Caspase3적표체。결과중리자조사가명현억제인외주혈T림파세포적증식，수착제량증대，억제작용경가명현。동시，중리자조사가촉진T림파세포적조망，특별시대우만기조망적유도작용（P＜0．01）。 RT-PCR검측결과현시，중리자복사가억제항조망단백Bcl-2적표체，촉진촉조망단백Bax화Caspase3적표체（P＜0．01）。결론중리자복사가현저영향억제T림파세포적증식병촉진기조망。
Objective To study the effect of heavy ion radiation on proliferation and apoptosis of human peripheral blood derived T lymphocytes and the mechanism .Methods T lymphocytes were isolated from heparinized whole blood samples by density gradient centrifugation using Ficoll before being irradiated with heavy ion beams 12 C.The accumulated absorbed dose (dose-rate values=0.5 Gy/min, and meanLET=29 keV/μm).12 h and 24 h post-infection, total RNA of T lymphocytes was isolated , and the apoptosis related gene expression , including Bcl-2, Bax, Caspase3, Caspase8 and Caspase9, was detected by RT-RT-PCR.24 h and 48 h after irradiation, the proliferation was analyzed by CCK 8 kit.The cell apoptosis was detected by flow cytometry after being labeled with AnnexinV-PE/7-AAD or AnnexinV-FITC/PE.The expression of Bcl-2, Bax and Caspase3 was also assayed by RT-PCR.Results Data showed that heavy ion radiation could inhibit the proliferation of T lymphocytes obviously , and the inhibition ratio in cells that received 2 Gy dose was much high-er than in cells that received 1 Gy dose.Furthermore, heavy ion radiation promoted the apoptosis of T lymphocytes signifi-cantly.The results of RT-PCR showed that the mRNA expression of Bcl-2 was down-regulated in heavy ion radiation T lym-phocytes while the expression of Bax and Caspase 3 was up-regulated.Conclusion Heavy ion radiation can inhibit the pro-liferation and promote the apoptosis of human peripheral blood derived T lymphocytes .