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目的 通过对宫颈癌细胞系HeLa细胞中Bcrp1+表型细胞的侵袭和成瘤能力的研究,探讨Bcrp1成为宫颈癌干细胞表面标志物的可能性.方法 根据实验中所用HeLa细胞表型的不同分为两组,即Bcrp1+组和Bcrp1 -组,利用穿膜小室——boydon小室体外侵袭实验检测两组细胞的侵袭能力；将不同细胞密度梯度(1×104、1×105、1×106个/ml)的Bcrp1+及Bcrp1-组细胞分别接种于6～8周龄非糖尿病-严重免疫缺陷(NOD/SCID)小鼠体内,比较两组小鼠的体内成瘤情况,包括成瘤时间、成瘤率、肿瘤体积及肿瘤质量.结果 体外侵袭实验显示,Bcrp1+组穿膜细胞数为(99± 14)个,侵袭深度为(366±52) μm,明显高于Bcrp1-组的(57±13)个和(301±54) μm,两组分别比较,差异均有统计学意义(P＜0.05).体内成瘤实验显示,接种细胞密度为1 ×104个/ml时,Bcrp1+ 组小鼠体内即可形成移植瘤；接种细胞密度为1×105和1×106个/ml时,Bcrp1+组和Bcrp1-组小鼠体内均可形成移植瘤；与Bcrp1-组小鼠比较,Bcrp1+组小鼠接种相应细胞密度的成瘤时间早、成瘤率高、肿瘤体积和肿瘤质量大,两组分别比较,差异均有统计学意义(P＜0.05).结论 Bcrp1+表型的HeLa细胞与Bcrp1-表型相比,具有较强的成瘤及侵袭能力,具备肿瘤干细胞的部分特征,其中可能富集了宫颈癌干细胞.

Objective To make sure whether or not Bcrp1 is the marker of cervical cancer stem cells or not by studying the invasive ability and formation of tumors of Bcrp1 + phenotype HeLa cells.Methods The tumor cell migration and invasion assay were used by boyden chamber to identify the invasive ability of Bcrp1 + phenotype HeLa cells.The formation of tumors in vivo experiments were completed,in which the two groups of cells with different concentrations were inoculated in non obese diabetes-severe combined immunodeficiency disease ( NOD/SCID ) mice ( 1 × 104,1 × 105,1 × 106/ml ) and the differences of time,rate and volume in the formation of tumors between two groups were observed.Results ( 1 ) In the invasion assay,the amount of cells that invaded through the artificial basement membrane in Bcrp1 + group were 99 ± 14,which was significantly greater than those in Bcrp1- group ( 57 ± 13,P ＜ 0.05 ) ; the length of the Bcrp1 + group was ( 366 ± 52 ) μm,which was significantly greater than the Bcrp1 - group ( 301 ± 54) μm ( P ＜ 0.05 ).( 2 ) Following transplantation of 1 × 104 cells,only the Bcrp1 + cells formed tumors in NOD/SCID mice.When 1 × 105 or 1 × 106 cells were transplanted,the tumor incidence and the tumor mass were greater in the Bcrp1 + groups than those in the Bcrp1 - groups ( P ＜ 0.05 ).Conclusion Bcrp1 + HeLa cell have the greater capacity of invasive and the tumorigenicity,which may contain cancer stem cells.