AIM： To evaluate the mechanism underlying the effects of 5-Fluorouracil （5-FU） on adenoviral infection. METHODS： Low and high Coxsackievirus-Adenovirus Receptor （CAR） expressing human colon carcinoma cell lines ...AIM： To evaluate the mechanism underlying the effects of 5-Fluorouracil （5-FU） on adenoviral infection. METHODS： Low and high Coxsackievirus-Adenovirus Receptor （CAR） expressing human colon carcinoma cell lines were treated with 5-FU and two El-deleted adenoviral constructs, one transferring GFP （Ad/CMV- GFP） the other bax （Ad/CEA-bax）. The number of infected cells were monitored by GFP expression. To evaluate the effects of 5-FU in a receptor free system, Ad/GFP were encapsulated in liposomes and treated with 5-FU. Ad/GFP release was estimated with PCR and infection of 293 cells with the supernatant. Electron microscopy of the Ad5-GFP-liposome complex was made to investigate morphological changes of the liposomes after 5-FU. RESULTS： Infection rates of all cell lines increased from 50% to 98% with emerging 5-FU concentrations. The enhanced viral uptake was independent of the CAR expression. Additionally, 5-FU treated liposomes released 2-2.5 times more adenoviruses. Furthermore, 5-FU- treated liposomes appeared irregular and porous-like. CONCLUSION： adenoviral uptake is enhanced in the presence of 5-FU irrespective of CAR and is associated with morphological changes in membranes making the combination of both a promising option in gene therapy.展开更多
文摘AIM： To evaluate the mechanism underlying the effects of 5-Fluorouracil （5-FU） on adenoviral infection. METHODS： Low and high Coxsackievirus-Adenovirus Receptor （CAR） expressing human colon carcinoma cell lines were treated with 5-FU and two El-deleted adenoviral constructs, one transferring GFP （Ad/CMV- GFP） the other bax （Ad/CEA-bax）. The number of infected cells were monitored by GFP expression. To evaluate the effects of 5-FU in a receptor free system, Ad/GFP were encapsulated in liposomes and treated with 5-FU. Ad/GFP release was estimated with PCR and infection of 293 cells with the supernatant. Electron microscopy of the Ad5-GFP-liposome complex was made to investigate morphological changes of the liposomes after 5-FU. RESULTS： Infection rates of all cell lines increased from 50% to 98% with emerging 5-FU concentrations. The enhanced viral uptake was independent of the CAR expression. Additionally, 5-FU treated liposomes released 2-2.5 times more adenoviruses. Furthermore, 5-FU- treated liposomes appeared irregular and porous-like. CONCLUSION： adenoviral uptake is enhanced in the presence of 5-FU irrespective of CAR and is associated with morphological changes in membranes making the combination of both a promising option in gene therapy.
