论文部分内容阅读
目的:用无血清培养基方法从人食管癌细胞株KYSE-150、TE-1中分离富含肿瘤干细胞的细胞球并鉴定其生物学特性。方法:运用无血清培养基培养KYSE-150、TE-1细胞,观察细胞球的生成及在有血清培养基中的分化情况,利用MTT法以及Transwell小室法研究食管癌细胞球的增殖情况和侵袭能力,流式细胞仪检测分子标志CD44、CD271的表达。结果:KYSE-150、TE-1细胞在无血清培养基培养条件下可以获得可稳定传代的细胞球,细胞球细胞的增殖能力和侵袭能力均高于其亲本细胞;无血清培养基培养下KYSE-150细胞球中CD44+、CD271+、CD44+CD271+细胞分别为(64.92±11.04)%、(28.27±6.79)%、(24.07±5.39)%,均显著高于亲本细胞[(37.04±6.30)%、(3.69±0.49)%、(1.64±0.11)%,P均<0.05]。TE-1细胞球中CD44+、CD271+、CD44+CD271+细胞占(6.41±0.87)%、(2.58±0.17)%、(0.27±0.53)%,均显著高于亲本细胞[(1.87±0.18)%、(0.44±0.10)%、(0.09±0.02)%,P均<0.05]。结论:应用无血清培养基可以从KYSE-150、TE-1细胞系中分离出具有干细胞特性的食管癌细胞球,CD44、CD271分子可能是食管癌干细胞的标志。
OBJECTIVE: To isolate and characterize tumor stem cell-rich cytoplasm from human esophageal cancer cell lines KYSE-150 and TE-1 using serum-free medium. Methods: KYSE-150 and TE-1 cells were cultured in serum-free medium. The formation of cell spheres and the differentiation in serum-containing medium were observed. The proliferation and invasion of esophageal cancer cell spheres were studied by MTT assay and Transwell chamber assay Capacity, flow cytometry detection of molecular markers CD44, CD271 expression. Results: KYSE-150 and TE-1 cells could obtain stably passaged cytoplasm under the condition of serum-free medium, the cell proliferation ability and invasive ability of cell were higher than that of their parental cells. KYSE- (64.92 ± 11.04)%, (28.27 ± 6.79)% and (24.07 ± 5.39)% in the -150 cell sphere were significantly higher than that of the parent cell [(37.04 ± 6.30)%, (3.69 ± 0.49)%, (1.64 ± 0.11)% respectively, all P <0.05]. The percentage of CD44 +, CD271 + and CD44 + CD271 + cells in TE-1 cells was (6.41 ± 0.87)%, (2.58 ± 0.17)% and (0.27 ± 0.53)%, (0.44 ± 0.10)%, (0.09 ± 0.02)%, P <0.05]. Conclusion: Stem cells can be isolated from KYSE-150 and TE-1 cell lines by using serum-free medium. CD44 and CD271 may be the hallmarks of esophageal cancer stem cells.