RAB27B在宫颈癌干细胞中的作用研究

Study on the role of RAB27B in cervical cancer stem cells

目的探讨RAB27B在宫颈癌干细胞中的作用,旨在为中晚期及复发患者提供新的治疗靶点。方法采用细胞转染的方法将si-NC、si-RAB27B片段成功转入宫颈癌SP细胞中;免疫蛋白印迹实验检测RAB27B敲除前后目的基因蛋白的表达情况;实时荧光定量聚合酶链式反应检测细胞目的基因的mRNA表达情况;体内功能实验检测RAB27B敲除前后细胞增殖、侵袭及克隆形成能力;免疫荧光检测RAB27B与SFRP1的细胞定位情况;酶联免疫吸附实验检测SFRP1分泌情况。结果敲除RAB27B后宫颈癌SP细胞中RAB27B的蛋白及mRNA表达明显降低;敲除RAB27B后宫颈癌SP细胞的增殖、侵袭及克隆形成能力明显下降;RAB27B与SFRP1在宫颈癌SP细胞中具有明显的共定位情况;敲除RAB27B后宫颈癌SP细胞分泌SFRP1的含量亦明显下降。结论RAB27B可以促进宫颈癌干细胞的增殖、侵袭及克隆形成,RAB27B可能是通过调控SFRP1表达而发挥作用。

Objective The aim of the stydy was to explore the role of RAB27B in cervical cancer stem cells and explore new therapeutic targets for patients with advanced and recurrent disease.Methods The si-NC and si-RAB27B fragments were successfully transferred into cervical cancer SP cells by cell transfection;Western blotting was used to detect the expression of target gene protein before and after RAB27B knockout;Real-time fluorescence quantitative polymerase chain reaction was used to detected the mRNA expression of target gene in cells;in vivo functional experiments to detect cell proliferation,invasion and clone formation before and after RAB27B knockout;immunofluorescence was used to detect cellular localization of RAB27B and SFRP1;enzyme-linked immunosorbent assay was used to detect SFRP1 secretion.Results After knocking out RAB27B,the protein and m RNA expressions of RAB27B in cervical cancer SP cells were significantly decreased;After RAB27B knockout,the proliferation,invasion and clone formation abilities of cervical cancer SP cells were significantly decreased.There was obvious co-localization between RAB27B and SFRP1 in cervical cancer SP cells after knocking out RAB27B;the content of SFRP1 secreted by cervical cancer SP cells was also significantly decreased.Conclusion RAB27B could promote the proliferation,invasion and clone formation of cervical cancer stem cells,and might work throughregulating the expression of SFRP1.