纳米板转导HPV18 E7siRNA对宫颈癌细胞增殖的抑制作用

Nanopatch-delivered HPV18 E7 siRNA inhibits proliferation of skin cells in cervical cancer transgenic mice

目的通过纳米板转导HPV18 E7 siRNA进入HPV18 E7宫颈癌转基因鼠皮肤组织,探讨纳米板转导siRNA的效率及转导的siRNA对宫颈癌细胞增殖的抑制作用。方法纳米板转导FITC标记的寡聚DNA(代替siRNA)进入小鼠皮肤,观察荧光变化,判断DNA是否被转导进入皮肤细胞中;纳米板转导DiI标记脂质体与siRNA复合物进入小鼠皮肤,测定转导前后和残留在皮肤上的荧光强度,计算纳米板转导siRNA进入组织细胞的效率;纳米板转导E7 siRNA和脂质体的复合物于转基因鼠皮肤中后,BrDU免疫组化检测siE7对宫颈癌细胞增殖的抑制作用。结果纳米板转导寡聚DNA后绿色荧光逐渐分散变多,表明DNA已逐渐进入小鼠皮肤细胞中。纳米板转导DiI标记脂质体与siRNA进入小鼠皮肤后,通过荧光强度计算得出转染率为50.23%。纳米板转导E7 siRNA进入HPV18 E7宫颈癌转基因鼠皮肤后,能有效抑制宫颈癌细胞增殖。结论纳米板能高效的转导siRNA进入组织细胞,并且转导进入细胞的siE7能有效抑制宫颈癌细胞增殖。

Objective To explore the efficiency of siRNA delivery by using nanopatch and the inhibi- tory effect of HPV18 E7 siRNA （siE7） on the proliferation of skin cells in cervical cancer transgenic mice. Methods The oligonucleotide tagged with FITC as a substitute of siRNA was delivered into mouse skin by nanopatch, and the green fluorescence was observed to analyze whether the nanopatch can deliver siRNA into the cells of mouse skin. Nanopatch was used to deliver the complexes of siE7 and liposomes marked with DiI into mouse skin. The fluorescence intensity of DiI before and after transfection and that remained on the surface of mouse skin after transfection were determined. The efficiency of siE7 delivery by nanopatch was calculated, and BrdU immunohistochemistry was applied to detect the proliferation of skin cells transfected with siE7 in cervical cancer transgenic mice. Results The green fluorescence detected by confocal laser scanning microscopy （CLSM） after transfection by nanopateh increased and scattered, suggesting the oligonucleotide was delivered into skin cells. The transfection efficiency of using nanopatch to deliver siRNA was 50.23 %. The proliferation of transgenic mouse skin cells transfected with siE7 by nanopatch was inhibited. Conclusion Nanopatch can effectively deliver siRNA into mouse skin ceils, and siE7 delivered by nanopatch can inhibit the proliferation of skin cells of cervical cancer transgenic mice.