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the use of vincristine (vcr) to treat cancer has been limited by its dose-dependent toxicity and development of drug resistance after repeated administrations. in this study, we investigated the mechanism by which quinine hydrochloride (qn) acts as a sensitizer for vcr. our experiments used three kinds of multidrug-resistant cancer cells and demonstrated that qn worked by inducing intracellular depletion of adenosine triphosphate, increasing adenosine triphosphatase activity, and decreasing p-glycoprotein expression. based on these results, we designed and prepared a vcr and qn codelivery liposome (vql) and investigated the effect of coencapsulated qn on the in vitro cytotoxicity of vcr in cells and three-dimensional multicellular tumor spheroids. the antitumor effects of the formulation were also evaluated in multidrug-resistant tumor-bearing mice. the results of this in vivo study indicated that vql could reverse vcr resistance. in addition, it reduced tumor volume 5.4-fold when compared with other test groups. the data suggest that vql could be a promising nanoscaled therapeutic agent to overcome multidrug resistance, and may have important clinical implications for the treatment of cancer.