Effects Of The Proliferation And Apoptosis Of AAVC-I On A549Cells And VEC Cells

Objective: To research the effects of the proliferation and apoptosis of anti-tumor component Ifrom Agkistrodon acutus venom(AAVC-I) on A549cells and VEC cells.Methods: VEC cells were extracted from neonate umbilical cord by enzyme digestion. It wasidentified by factor Ⅷ-related antigen immunohistochemical staining. The concentrationgradients of AAVC-I was2.5,5,10,20,40μg/ml. The inhibition rate after12h,24h and48h withdifferent concentrations of AAVC-I on A549and VEC was detected by MTT method. Themorphological change of A549and VEC was observed by HE staining, AO/EB fluorescentstaining and Hoechst33258fluorescent staining. Condition apoptosis after AAVC-I treatmentwas detected with FCM. The change of bax and caspase-3expression after AAVC-I treatmentwas detected by immunohistochemical staining method.Results: VEC cells grow adhering to the wall and forming a monolayer after fusion. It willspread to more than80%of the bottom of the culture bottle after6-8days, and it will arrayedinto a cobblestone-like structure. After immunohistochemical staining, the positive staining oncells can be observed, which showed that it is VEC cells.MTT: The proliferation of A549and VEC was inhibited by AAVC-I in both time anddose-dependent manners.HE staining, AO/EB fluorescent staining and Hoechst33258fluorescent stainingshowed the typical features of apoptosis on A549and VEC after AAVC-I treatment for24h.FCM: With different concentration of AAVC-I treatment for24h, the apoptosis rate ofA549increased form0.64%to70.97%, and the apoptosis rate of VEC increased form0.47%to78.81%. Immunohistochemical staining: Compared with control group, the positive staining canbe observed in experimental group. With the increase of AAVC-I concentration, the mean lightdensity of A549and VEC have a trend of increase, which indicate the expression of bax andcaspase-3increase.Conclusion:1. AAVC-I could inhibit the proliferation of A549and VEC.2. AAVC-I could induce the apoptosis of A549and VEC3. The pro-apoptotic effect of AAVC-I on A549and VEC was related to theexpression of bax and caspase-3...