摘要
目的构建pEgr-Endostatin-EGFP重组质粒,研究其体内、体外X射线诱导表达特性和对肿瘤生长的抑制作用。方法构建pEgr-Endostatin-EGFP重组质粒,脂质体包裹转染B16细胞系,通过RT-PCR方法检测Endostatin表达特性,流式细胞术法检测EGFP表达特性。建立荷瘤小鼠模型,给予质粒+5 Gy X射线治疗,共3次,检测18日肿瘤组织重量。结果真核表达载体pEgr-Endostatin-EGFP构建成功并转染B16细胞。被转染的B16细胞经0.05-20 Gy X射线照射,EGFP表达明显增高。2 Gy照射后8 h EGFP表达达到最高值。体内研究证实,pEgr-Endostatin-EGFP基因-放射治疗能够明显抑制肿瘤组织生长。结论pEgr-Endostatin-EGFP重组质粒具有辐射诱导表达增强的特性,pEgr-Endo-statin-EGFP基因-放射治疗具有显著的抗黑色素瘤作用,本实验结果为肿瘤基因-放射治疗研究的发展提供了实验依据。
Objective pEgr-Endostatin-EGFP plasmid was constructed to investigate its expression properties induced by ionizing irradiation invitro and invivo. Methods The reconrbined plasmid pEgr-Endostatin-EGFP was constructed and transfected into B16 cell line with liposome. The expression property of Endostatin was investigated by RT-PCR and that of GFP was detected by FACS analy- sis .Then, tumor-bearing model was establislied treated by plasmids injection and 5Gy X-ray irradiation for 3 times. Tumors＇ weiglit were detected 18 days later.Results The expression of GFP in B16 melanoma cells was detected after X-irradiation with 0.05 - 20 Gy. Time-course studies sliowed that the expression of GFP in B16 cells reached its peak at 8 h after irradiation with 2 Gy.The injec- tion of pEgr-Endostatin-EGFP recombinant plasmid into implanted B16 melanoma in C57BL/6J mice followed by local X-irradiation could significantly inhibit tumor growth. Conclusion The anti-tumor effect of pEgr-Endostatin-EGFP gene- radiotherapy was most prominent for melanoma, the results would provide an experimental basis for planning effective clinical gene-radiotherapy of cancer.
出处
《中国实验诊断学》
2007年第10期1281-1284,共4页
Chinese Journal of Laboratory Diagnosis
