Abstract
Objective To study the effect of as - miR - 21 enhance U251 human glioma cell chemo-sensitivity to 5 - FU. Method 5 - FU/PAMAM complex was prepared by dialysis method. As - miR - 21 was incubated with 5 - FU/PAMAM to form 5 - FU/PAMAM/as - miR - 21. Transmission electronic microscopy (TEM) was performed to observe the morphology of the nanoparticles. The drug loading efficiency and encapsulation efficiency was determined by ultraviolet spectroscopy ( UV ). The transfection of PAMAM dendrimer was detected by flow cytometry assay. MTT assay was carried out to determine U251 cell growth survival rate. Cell apoptosis was analyzed by flow - cytometry assay. Immunofluoresence staining was employed to determine Ki67 and Bcl -2 expression in U251 cells after combination treatment Transwell assay was performed to detect cell invasion ability. Results The morphology of the nanoparticle was sphere observed by TEM, and the diameter was less than 100 nm. The encapsulation efficiency and loading efficiency of drug were determined by ultraviolet spectroscopy to be 66. 21% and 31. 77% ,respectively. The results of flow cytometry assay showed that PAMAM dendrimer transfection efficiency was 70. 53%. The survival rate of U251 cells with as - miR - 21 and 5 - FU co - delivery treatment was significantly suppressed(F=273.345,P=0.000). The increased apoptosis nuclei percentage in co - delivery treated U251 cell was detected ( F =43. 21, P =0. 000) by flow - cytometry assay and the expression of Ki67 and Bcl -2 were down regulated simultaneously analyzed by immunofluorescence. Transwell result also demonstrated the cell invasion ability of U251 cells treated with 5 - FU combine with as - miR - 21 was decreased. Conclusions PAMAM dendrimer could effectively deliver as - miR - 21 and 5 - FU simultaneously;combination therapy can suppress U251 cell line growth effectively in vitro and can enhance the chemo -sensitivity of glioma cells to 5 - FU chemotherapy. Key words: PAMAM; Co-delivery; As-miR-21; Fluorouracil; Glioma
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