A Study Of Anti-NGPT2 Antibody For The Treatment Of AML In Mouse Model

Objective:To understand the effect of anti-ANGPT2 on the AML mouse model, construct an AML mouse model was established. The downstream signal expressed downstream was detected in tumor tissues with immunohistochemical method. Various expressions in tumor tissue, vascular proliferation and survival duration of mouse were observed. This study may be used as open a new way for the treatment of acute myeloid leukemia.Methods:1、 A logarithmic growth phase-HL60 cell line was inoculated in NOD/SCID mouse for the establishment of acute myeloid leukemia in mouse model.2、Intraperitoneal injection of ANGPT2, VEGF monoclonal antibody, monoclonal antibody IgG and physiological saline to be injected in the mouse for the study of effect on the AML.3、To observe the survival period and tumor growth in mouse.4、To dissect the anatomical model of mouse by using western blot method to detect the TIE2/SHP2/DOKR/PI3K protein of the tumor tissue. Using immunohistochemical method to detect the tumor angiogenesis and Ki-67 for the proliferation of tumor tissue.5、Application of Spearman correlation analysis for the relation between tumor associated protein expression and the survival period of mouse.Results:1) NOD/SCID AML mouse was established successfully.2) The application of anti-ANGPT2 mAb and anti-ANGPT2 mAb+ anti -VEGF mAb in acute myelogenous leukemia mouse showed that anti-ANGPT2 mAb and anti-ANGPT2 mAb+anti-VEGF mAb mouse tumor volume and IgG group was significantly difference compared with the control group. A longer survival time was observed. At the same time, there were no leukemia cells in mouse liver, spleen, lung and brain. The western blot method was used to detect the expression of downstream protein in tumor tissues. The expression of TIE2 was negative whereas SHP2 DOKR/PI3K protein expression was positive. The detection of CD31 MVD in tumor tissues was negative while using CD34, which was still negative. The degree of proliferation of tumor tissues by using Ki-67 was above 90%.Conclusions:1、Adequate doses of anti ANGPT2 antibody for the treatment of acute myeloid leukemia in mouse can prolong the survival and improve the quality of life in mouse model.2、Due to the treatment effect, Ki-67 was strongly expressed, that may be used to evaluate the prognosis of AML. At the same time, the western blot test was positive in downstream signal protein. This may provide a new idea for future research of leukemia.