Zhao Yongbo, Wang Feng, Li Yu, Zhang Ying, Zhang Guiyin
Department of Neurology, Shanghai First People's Hospital, Shanghai 200080 P. R. China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2002 Apr;19(2):130-3.
To detect the expressing levels of human tissue-plasminogen activator(t-PA) in AGZY83-a cells transfected with pcDNA3.1(+) t-PA in vivo and the feasibility of using transplantation of cells for gene therapy of thrombotic diseases.
Expression vectors containing the t-PA cDNA gene were transfected into AGZY83-a cells. The transfected AGZY83-a cells were implanted into mice in different regions, and the plasma levels of human t-PA were assayed at intervals.
The plasma levels of human t-PA were significantly increased in mice after implantation of transfected AGZY83-a cells and were significantly higher than those of control groups implanted with untransfected AGZY83-a cells. This significant increase lasted at least 105 days. The intraperitoneal implantation group expressed the highest level of human t-PA, a little higher than that of the subcutis implantation group, while both are much higher than that of the quadriceps femoris implantation group which expressed the lowest.
The implanted transfected AGZY83-a cells are able to stably express high levels of human t-PA, and transplantation of cells transfected with pcDNA3.1(+) t-PA is a new promising method for gene therapy of thrombotic diseases.
检测经pcDNA3.1(+)t-PA转染的AGZY83-a细胞在体内人组织型纤溶酶原激活剂(t-PA)的表达水平以及细胞移植用于血栓性疾病基因治疗的可行性。
将含t-PA cDNA基因的表达载体转染至AGZY83-a细胞。将转染后的AGZY83-a细胞植入小鼠的不同部位,并定期检测人t-PA的血浆水平。
植入转染后的AGZY83-a细胞的小鼠血浆中人t-PA水平显著升高,且明显高于植入未转染AGZY83-a细胞的对照组。这种显著升高至少持续105天。腹腔植入组人t-PA表达水平最高,略高于皮下植入组,而两者均远高于表达水平最低的股四头肌植入组。
植入的转染AGZY83-a细胞能够稳定高效表达人t-PA,经pcDNA3.1(+)t-PA转染的细胞移植是一种有前景的血栓性疾病基因治疗新方法。