Construction of an adipose derived stem cell-based alphastatin peptide glioma targeting vector and its inhibitory effect on angiogenesis / 西安交通大学学报(医学版)

ABSTRACT

【Objective】 To construct an adipose derived stem cell-based alphastatin peptide glioma targeting vector and detect its anti-angiogenesis effect in vitro. 【Methods】 The adipose derived stem cell-based alphastatin peptide glioma targeting vector (Al-ADSCs) was constructed by transfecting the alphastatin peptide lentivirus vector into adipose derived stem cells (ADSCs). The expressions of stem cell markers on the surface of targeted vector were detected by flow cytometry. The expression of alphastatin peptide in the targeted vector and in the cell culture supernatant of the targeted vector were detected by Western blotting and ELISA, respectively. Cell migration assay was used to detect the tendency of the targeted vector toward CD133+ glioma stem cells, and lumen formation assay was used to detect the effect of the targeted vector on endothelial cell angiogenesis in vitro. 【Results】 After transfection, the surface markers of stem cells expressed by targeted vector did not significantly change compared with ordinary adipose derived stem cells. Western blotting showed that the targeted vector could successfully express alphastatin peptide. ELISA showed that the alphastatin peptide was detected in the cell culture supernatant of targeted vector \mg/L]. Cell migration test showed no significant difference in the tendency of CD133+ glioma stem cells between the targeted vector and ordinary adipose derived stem cells \. Lumen formation experiment showed that the targeted vector could inhibit endothelial cell-mediated angiogenesis in vitro [Lumen count Control group (13.33±0.76)/HPF, ADSCs group (19.40±1.71)/HPF, Al-ADSCs group (7.27±0.31)/HPF, P<0.01]. 【Conclusion】 In the process of constructing the adipose derived stem cell-based alphastatin peptide glioma targeting vector, the stem cell biological characteristics and tumor tendency of targeted vector have no significant changes. This targeted vector can stably express and secrete alphastatin peptide and inhibit endothelial cell-mediated angiogenesis in vitro.