Anti-Carcinoma Effect Of Folic Acid-Chitosan-Carbon Nanotube-Doxorubicin On Tumor Growth Of SMMC-7721 Liver Cancer And Its Toxicity In Nude Mice

Objective:In order to increase the antitumor effect of doxorubicin, we investigated the in vitro and in vivo anticarcinoma effect of Folic acid-Chitosan-Carbon nanotube-Doxorubicin (DOX-FA-CHI-SWCNT) on SMMC-7721 liver cancer and its toxicity in nude mice.Methods:FA-CHI-SWCNT, DOX-FA-CHI-SWCNT and DOX were incubated with SMMC-7721 cells for 24h, along with control group were analyzed by fluorescence microscopy after stained by AO/EB. SMMC-7721 cells were incubated with different dose of SWCNT, CHI-SWCNT, FA-CHI-SWCNT, CHI-SWCNT-DOX, DOX-FA-CHI-SWCNT and DOX, then the cell viability at 24h,48h and 72h was detected respectively using the WST-1 assay. The SMMC-7721 bearing nude mice were randomly allocated into 7 treatment groups including SWCNT, CHI-SWCNT, FA-CHI-SWCNT, CHI-SWCNT-DOX, DOX-FA-CHI-SWCNT, DOX and NS control group. They were exposed by intravenous injection respectively at day 1 and day 6. The general conditions and weights of the animals were observed, the length and width of the tumors were tested and the tumor volume and the inhibition rates of tumor growth were calculated. Then the mice were sacrificed at d21, blood routine, serum biochemistry parameters including alanine aminotransferase (ALT), aspartate aminotransferase (AST) and creatinine (Cr) were all detected. At last, the pathological changes of tumor, liver and kidney were observed under light microscopy.Results:Fluorescence microscopy and cell viability results showed that the cytotoxicity of drugs on SMMC-7721 cells apparently followed a sequence order:DOX-FA-CHI-SWCNT> CHI-SWCNT-DOX> DOX> FA-CHI-SWCNT. DOX-FA-CHI-SWCNT exhibited more effective cytotoxicity and exhibited time-effect and dose-effect relationship as well. The general situation of nude mice indicated that SWCNT and CHI-SWCNT were more toxic, which could cause death and weight loss. The changes of tumor volume demonstrated that DOX-FA-CHI-SWCNT could inhibit the growth of SMMC-7721 liver cancer, possessed a stronger antitumor effect than other drugs and showed slow-release characteristic. The histopathological results showed there was no significant difference on tumor necrosis in each group. Blood routine and serum parameters of nude mice suggested DOX could cause platelet (PLT) and aspartate aminotransferase (AST) levels significantly increased, while DOX-FA-CHI-SWCNT caused less change. Histopathologic photo in DOX group showed liver cell necrosis and inflammatory cell infiltration in portal areas, which pathological changes were more serious compared with other drugs. There was no significant change in renal histology in DOX and DOX-FA-CHI-SWCNT group.Conclusions:Compared with DOX, DOX-FA-CHI-SWCNT possessed more efficient cytotoxicity on SMMC-7721 cells and exhibited time-effect and dose-effect relationship as well. In vivo experiment demonstrated that DOX-FA-CHI-SWCNT could inhibit the growth of SMMC-7721 liver cancer more effectively than other drugs and resulted in less toxicity in nude mice than DOX.