中国现代医学杂志2018，Vol.28Issue(17)：6-12,7.DOI:10.3969/j.issn.1005-8982.2018.17.002
分选卵巢癌腹水中癌细胞的实验研究
Establishment of methodology of sorting tumor cells from ovarian cancer peritoneal fluids
摘要
Abstract
Objective To investigate the viability of the protocol to sort tumor cells from ovarian cancer peritoneal fluids by combining immunocytochemistry (ICC) and fluorescence in situ hybridization (FISH). Methods The ovarian cancer peritoneal fluid model was established and divided into a negative control group and four positive groups by adding 0, 5, 10, 20 and 40 ovarian cancers SKOV3 cells marked by Mito-Tracker Green into 4 ml benign peritoneal washing fluid respectively, 3 duplications per group. The nude mouse model bearing orthotopically transplanted human ovarian cancer SKOV3 cells was established, including the negative control group and the positive group, with 6 BALB/c nude mice per group and 2 nude mice at each time point. Peritoneal fluids of the nude mice in the positive group were collected at the 4th, 6th and 8th w after transplanting SKOV3 cells under the capsule of the left ovary. SKOV3 cells were enriched from the peritoneal fluids by magnetic activated cell sorting (MACS). Then, the cancer cells were identified exactly by EBA-1-ICC and CEP8-FISH (Chromosome Enumeration Probe). The cells with the characteristic of DAPI+/EBA-1+/Mito-Tracker Green+/CEP8+ were classified as detected cancer cells by the positive standard of CEP8 >2. Results In the peritoneal fluid model, the EBA-1-positive cells were found in the negative control group and all SKOV3 cell-positive groups, the marked SKOV3 cells were found in all SKOV3 cell-positive groups and their recovery rates were 20%-50%, the detection rates in 9 of 12 SKOV3 cell-positive samples were 100%. In the nude mouse model, under every 20×10 microscopic field, the maximum number of SKOV3 cancer cells was 3, 8 and 15 respectively at the 4th, 6th and 8th w. Conclusions The protocol of combining ICC and FISH can exactly identify cancer cells from ovarian cancer peritoneal fluids which have been enriched by MACS. The technique provides new ideas for studying ovarian cancers.关键词
卵巢癌/腹水/癌细胞/免疫细胞化学/荧光原位杂交Key words
ovarian cancer/ascites/cancer cell/immunocytochemistry/fluorescence in situ hybridization分类
医药卫生引用本文复制引用
邢宝玲,郭存存,唐正华,董一善,葛素梅..分选卵巢癌腹水中癌细胞的实验研究[J].中国现代医学杂志,2018,28(17):6-12,7.基金项目
江苏省常州市科技计划项目(No:CJ20159040) （No:CJ20159040）
