Killing effect of immunoliposomal adriamycin on breast cancer cells in vitro

Abstract

Objective To construct the immunoliposomal adriamycin tagged with trastuzumab F (ab') 2 and to investigate its killing effect on human breast cancer cells in vitro.Methods The tastuzmab F(ab')2 and the liposomal adriamycin were linked together by antibody crosslinking method.The morphology of immunoliposomes was observed by transmission electron microscopy (TEM) and their size distribution was measured by graphics software.The immune activity of immunoliposomes was evaluated using the fluorescence microscopy after the fluorescent immunoliposomes or lipoosomes were incubated together with MCF-7 cells.Inhibitory effect of the cell growth at different drug concentrations was measured using methyl thiazol tetrazolium (MTT) 8,16,24 and 32 h after cell culture.The apoptosis was examined by using flow cytometry 32 h after cell culture.Results The immunoliposomal adriamycin tagged with trastuzumab F(ab')2 that was constructed in vitro has an average diameter of 192 nm,and that the diameters under 200 nm accounted for 92.20％ of the total.The immunoliposome was superior to liposome at the identification capability to MCF-7 cells,the proportions of the cells which intake drug were (19.39 ±3.33)％,(57.36 ±4.12)％,(89.72 ±-5.88)％ and (10.17 ±4.05)％,(25.15 ± 3.77)％,(38.86 ±6.24)％ (P＜0.05) in two groups respectively after 10,30 and 50 min cell cultrue.The immunoliposome could inhibit the proliferation of MCF-7 cells in a dose (F =1 287.472,P ＜ 0.01) and time (F =2 145.325,P＜ 0.01)dependent manner; Apoptotic index of MCF-7 cells in immunoliposome group (35.22 ± 3.52) ％ was obviously higher than that in the control groups (10.31 ± 4.62) ％ 32 h after transfection.(P ＜ 0.05).Conclusion The immunoliposomal adriamycin tagged with trastuzumab F (ab') 2 was constructed successfully and it can inhibit the proliferation of kill the MCF-7 cells effectively in vitro. Key words: Immunoliposome; Adriamycin; Breast cancer