Studies On Antitumor Activity Of Vinblastine-loaded Folate-conjugated Albumin Nanoparticles

In recent years,with the rapid development of economy,society and intensify aging of population,in China,the morbidity and mortality of malignant tumor is on the rise.Cancer caused serious harm to people’s health and social development,thus cure cancer has become the pharmaceutical and chemical workers’ urgent task.Vinblastine exists in catharanthus roseus.As a cell cycle specific anti-tumor drug,vinblastine has been widely used in most of the cancer treatment.However,there are some disadvantages such as side effects on body,local stimulation,short half-life and no target objecet,make vinblastine restricted in the clinical applications.In this paper,we do anti-tumor experiments of FA-VLB-BSANP in vivo and in vitro on pharmacological evaluation index such as anti-tumor activity and anti-tumor mechanisms,so as to provide theoretical basis for the research of vinblastine new dosage forms.1.The anti-tumor activities of FA-VLB-BSANP in vitro was analyzed by MTT assay.The results showed that SGC-7901 cells were the most sensitive towards VLB and FA-VLB-BSANP,and the IC50 value of SGC-7901 was 69.19±7.32μg/mL and 60.63±8.38μg/mL after 48h treatment.And VLB and FA-VLB-BSANP inhibited the growth of SGC-7901 cells in a time and dose-dependent manner.Through the cell growth curve,we can see that the inhibition of cell proliferation ability of FA-VLB-BSANP was significantly stronger than that under VLB,and the inhibitory effect became more obvious with the time prolonged.Moreover,the morphological changes of cells were observed by using inverted microscope.It was showed that the effect on the cell proliferation is much stronger by using FA-VLB-BSANP when contrast to VLB.2.Apoptotic population of SGC-7901 cells was analyzed by flow cytometry via using AnnexinV-FITC/PI.The apoptotic results showed that the apoptotic population increased in a concentration-dependent manner with increasing concentration after the cells treated with VLB and FA-VLB-BSANP.Under the same dose and time conditions,the FA-VLB-BSANP presented better activity for inducing SGC-7901 cells apoptosis comparing with VLB.The cells morphological changes were observed by inverted fluorescence microscope via using acridine orange.It was showed that under the same dose and time conditions,the FA-VLB-BSANP presented better activity on inducing SGC-7901 cells apoptosis compared with VLB.3.We tested VLB and FA-VLB-BSANP induce the alterations in mitochondrial membrane potential of SGC-7901 cells by flow cytometry and inverted fluorescence microscope.The results showed that after treated with VLB and FA-VLB-BSANP in low fluorescence,the percentage of cells produced a time-dependent increase.And the percentage of cells treated with FA-VLB-BSANP in low fluorescence is much higher than those treated with VLB.Fluorescence microscope analysis showed that under the same dose and time conditions,the effect of FA-VLB-BSANP induce mitochondrial membrane potential decrese was significantly increased compared with VLB.4.ELISA was used to measure the levels of related proteins.The results showed that FA-VLB-BSANP has significant effect on Bcl-2 protein down regulation and up regulation of Bax protein than that treated by VLB.Besides,the expression of cyclinB1 and destructive effects on microtubule macrophage are both obvious than using VLB,blocked more tumor cells on M stage.In addition,comparing with VLB,FA-VLB-BSANP had more significant regulation effects on the IκB-α protein down regulation and up regulation of MEKK1 protein.5.ELISA was used to measure the effect on the SGC-7901 cells’ability of invasion and metastasis.The results showed that FA-VLB-BSANP had more significant inhibitory effects on the expression of MMP-2、MMP-9 and VEGF than VLB.6.The mice bearing tumors of S180 were selected to evaluate the anti-tumor activity in vivo of VLB and FA-VLB-BSANP.The results showed that the median lethal dose of VLB was 24.69mg/kg,the median lethal dose of FA-VLB-BSANP was 28.61mg/kg,which means the in vivo toxicity of FA-VLB-BSANP was significantly reduced.Inhibition rate of tumor growth were 79.43% and 86.86% at a dose of 4mg/kg,showed that FA-VLB-BSANP exhibited better anti-tumor activity.In summary,both anti-tumor activity in vitro and in vivo studies were shown that FA-VLB-BSANP had better anti-tumor activity than VLB,improved the inhibiton of tumor cells and the ability of invasion and metastasis,along with significantly reduction of toxicity in vivo.In conclusion,this study suggests that FA-VLB-BSANP is a novel formulation with high anti-tumor efficacy,low toxicity,and has an excellent potential to be developed as a new agent for cancer therapy.