Conduction of a prostate cancer whole-cell vaccine modified by mice HSP70 and GM-CSF
- VernacularTitle:鼠源性HSP70、GM-CSF双修饰前列腺癌全细胞疫苗的建立与鉴定
- Author:
Qi MA
1
;
Junfeng CHEN
;
Junhui JIANG
;
Ping WANG
;
Zhaohui JIANG
;
Yue CHENG
Author Information
1. 浙江大学宁波医院 宁波市第一医院泌尿外科
- Keywords:
Heat-shock protein;
Granulocyte macrophage colony-stimulating factor;
Prostate cancer whole-cell vac-cine
- From:
China Modern Doctor
2015;(19):1-4,11
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a whole-cell vaccine modified by mice HSP70 and GM-CSF in prostate cancer. Methods Sequence fragments carrying mice Hspa1a coupled with C-myc or Csf2 coupled with V5 tag were constructed by chemical synthesis and PCR amplification , then both fragments were cloned into the vector pMD-18T. After se quencing, fragments described above were cloned into pLenti6.3_MCS lentiviral vector. The lentiviral expression vectors were named as pLenti6.3_MCS_Hspa1a_cMyc and pLenti6.3_MCS_Csf2_V5. These vectors were transfected or co-transfected into mice prostate cancer cell line RM-1 and proteins were detected by Western blotting. Following irradia-tion of 10 Gy, cells lost proliferative ability and were frozen and then stored. Results Recombinant lentiviral vectors pLenti6.3_MCS_Hspa1a_cMyc and pLenti6.3_MCS_Csf2_V5 were successfully constructed and GM-CSF, HSP70 or both expression in the transfected RM-1 cells were verified by Western blotting. Conlusion The RM-1 whole-cell vaccines modified by HSP70 and GM-CSF have been successfully constructed, which facilitate further investigation of the action mechanisms of whole-cell antigen and immune enhancement.
