论文部分内容阅读
目的研究人胃癌BGC-823肿瘤细胞系中是否存在悬浮生长的肿瘤球,检测前胃泌素(progastrin,Pro-GRP)在肿瘤球中的表达。方法取贴壁生长的BGC-823细胞,用有限稀释法和无血清培养法培养,并利用软琼脂克隆形成实验证实BGC-823中存在悬浮生长的具有干细胞特性的肿瘤球,流式细胞仪(FCM)检测该肿瘤球中ABCG2、CD44、CD166的表达。ELISA、细胞免疫荧光、FCM方法比较血清Pro-GRP分别在肿瘤球和贴壁细胞中的表达差异。结果有限稀释法和无血清培养法均在BGC-823细胞系中培养出了悬浮生长的肿瘤球;克隆形成能力显示:肿瘤球为(37.63±5.99),贴壁细胞为(10.75±3.37),二者间有统计学差异(P<0.01);FCM显示肿瘤球ABCG2的表达率(57.17±3.49)明显高于贴壁细胞(8.67±0.77)(P<0.01);血清Pro-GRP在肿瘤球中的表达(73.37±9.42)明显高于贴壁细胞(5.11±1.03)(P<0.01)。结论 人胃癌BGC-823细胞系中存在悬浮生长的肿瘤球,Pro-GRP的表达与肿瘤球有关。
Objective To investigate the existence of tumor-growth spheres in human gastric cancer cell line BGC-823 and the expression of progastrin (Pro-GRP) in tumor spheres. Methods Adherent BGC-823 cells were cultured in a limited dilution method and serum-free medium, and the stem cells with suspension growth in BGC-823 were confirmed by soft agar colony formation assay. Flow cytometry FCM) to detect the expression of ABCG2, CD44, CD166 in this tumor sphere. ELISA, immunofluorescence and FCM were used to compare the expression of Pro-GRP in tumor spheres and adherent cells respectively. Results The suspension-grown tumor spheres were cultured in BGC-823 cells by both limiting dilution and serum-free culture. The clonogenic capacity of tumor cells was (37.63 ± 5.99) and (10.75 ± 3.37) (P <0.01). FCM showed that the expression of ABCG2 in tumor spheres (57.17 ± 3.49) was significantly higher than that in adherent cells (8.67 ± 0.77) (P <0.01) (73.37 ± 9.42) was significantly higher than that of adherent cells (5.11 ± 1.03) (P <0.01). Conclusions There are tumor spheres growing in BGC-823 cell line. The expression of Pro-GRP is related to tumor spheres.