转铁蛋白修饰多柔比星脂质体靶向抑制乳腺癌细胞增殖的研究

目的：利用转铁蛋白（TF）修饰多柔比星脂质体，并探讨其对乳腺癌细胞增殖的影响。方法采用薄膜超声法制备脂质体，利用硫酸梯度法制备多柔比星脂质体，然后采用后插入法制备 TF修饰多柔比星脂质体。激光共聚焦显微镜检测乳腺癌细胞 MCF-7和 MDA-MB-231对 TF-多柔比星脂质体的摄取，四甲基偶氮唑盐比色法（MTT 法）检测 TF-多柔比星脂质体靶向作用 MCF-7和 MDA-MB-231细胞后的杀伤能力，软琼脂克隆集落实验检测 TF-多柔比星脂质体对 MCF-7和 MDA-MB-231细胞的生长抑制作用。结果激光共聚焦显微镜观察显示，MCF-7细胞和 MDA-MB-231细胞对 TF-多柔比星脂质体摄取率显著高于多柔比星脂质体；MTT 法检测结果则显示，TF-多柔比星脂质体对 MCF-7细胞[（20．8±3．2）μmol／L]和 MDA-MB-231细胞[（20．1±3．0）μmol／L]杀伤的 IC50显著低于多柔比星脂质体[（158．6±24．6）μmol／L；（160．1±25．1）μmol／L）]和游离多柔比星[（161．7±26．2）μmol／L；（166．9±27．0）μmol／L）]，差异有统计学意义（F ＝116．03，P ＜0．001；F ＝75．29，P ＜0．001）。软琼脂克隆集落实验显示，TF-多柔比星脂质体对克隆集落的生长抑制显著优于多柔比星脂质体、游离多柔比星及对照组[MDA-MB-231细胞直径：（60．5±10．4）μm、（94．3±16．8）μm、（131．8±22．6）μm、（162．8±30．3）μm；MCF-7细胞直径：（31．8±5．5）μm、（62．1±11．1）μm、（108．6±18．6）μm、（157．4±29．3）μm]，差异有统计学意义（F ＝87．17，P ＜0．0001；F ＝178．23，P ＜0．0001）。结论 TF-多柔比星脂质体对乳腺癌细胞体外增殖具有显著的抑制作用，能够高效、特异地杀死乳腺癌细胞，为体内治疗乳腺癌提供了一定的理论依据。

Study of transferrin modified doxorubicin liposome targeted to inhibit proliferation of breast cancer cells

Objective To modified doxorubicin liposome with transferrin(TF),and to investigate its inhibition efficacy on the proliferation of human breast cancer cells.Methods The liposome was prepared by thin film ultrasonic,and doxorubicin liposomal was prepared by sulfuric acid gradient.The TF-doxorubicin lipo-some was prepared by the post insertion method.The uptake of TF-liposomal doxorubicin on breast cancer cells MCF-7 and MDA-MB-231 were detected by confocal microscopy.The killing ability of TF-doxorubicin liposomal targeting for MCF-7 and MDA-MB-231 were detected by MTT assay.Inhibitory effect of TF-doxorubicin lipo-some on the growth of MCF-7 and MDA-MB-231 were detected by soft agar colony assay.Results Confocal microscopy result showed that the uptake of TF-liposomal doxorubicin on MCF-7 and MDA-MB-231 were signifi-cantly higher than doxorubicin liposomal.Cell-killing ability on MCF-7 and MDA-MB-231 showed that the IC50 in TF-liposomal doxorubicin [MCF-7 cells:(20.8 ±3.2)μmol/L;MDA-MB-231 cells:(20.1 ±3.0)μmol/L)] were significantly lower than the liposomal [(1 58.6 ±24.6)μmol/L;(1 60.1 ±25.1 )μmol/L)]and free doxorubicin [(1 61 .7 ±26.2)μmol/L;(1 66.9 ±27.0)μmol/L)],with significant differences(F =1 1 6.03, P <0.001 ;F =75.29,P <0.001 ).Soft agar colony assay showed that the inhibition of TF-doxorubicin lipo-some on colony growth were significantly higher than doxorubicin liposome,free doxorubicin and control [dia-meter of MDA-MB-231 cells:(60.5 ±10.4)μm,(94.3 ±16.8)μm,(1 31 .8 ±22.6)μm,(162.8 ±30.3)μm;diameter of MCF-7 cells:(31 .8 ±5.5)μm,(62.1 ±11 .1 )μm,(108.6 ±1 8.6)μm,157.4 ±29.3)μm],with significant differences (F =87.17,P <0.000 1 ;F =178.23,P <0.000 1 ).Conclusion TF-doxorubicin lipo-some has a significant inhibitory effect on the proliferation of breast cancer cells in vitro,and can effectively and specifically kill the breast cancer cells,which provides theoretical basis for the treatment of breast cancer in vivo.