食管癌细胞株中肿瘤干细胞样细胞的分离培养及生物学鉴定

Isolation and biological characteristics of cancer stem cells from esophageal cancer cell lines

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 establish a culture method of cancer stem cells （CSCs） from esophageal cancer cell lines KYSE-150 and TE-1 by serum-free medium（SFM） and identify their characteristics. Methods：KYSE-150 and TE-1 cells were cultured in serum-free medium. The cell spheres were observed and cell differentiation was induced by serum supplement. MTF assay and invasive assay were applied to examine the invasive ability of both cell spheres and their parental cells. CD44＋ ,CD271 ＋ ,CD44＋CD271 ＋ tumor spheroid cells were also detected by flow cytometry. Results： After SFM culture, KYSE-150 and TE-1 cells formed spheres,and could be passaged continuously. The differentiation of cell spheres recurred when serum was supplemented. The proliferation and invasive ability of cell spheres were higher than their parents cells. CD44＋, CD271＋, CD44＋CD271＋ in KYSE-150 cell spheres were （64.92 ± 11.04）%, （28.27 ± 6.79）%, （24.07 ± 5.39）%,which were significantly higher than those in parents cells [ （37.04 ± 6.30）%, （3.69 -± 0.49）%, （1.64 ± 0.11）%,P 〈 0.05]. CD44＋,CD271＋,CD44＋CD271＋ TE-1 cell spheres were （6.41 ± 0.87）%, （2.58 ± 0.17）%, （0.27 ± 0.53）%,which were also significantly higher than those in parents cells [（1.87 ± 0.18）%, （0.44 ± 0.10）%, （0.09 ± 0.03）%,P 〈 0.051. Conclusion：Esophageal cancer cell spheres can be isolated from KYSE-150 and TE-1 by culture with SFM. CD44 and CD271 may be the cell surface makers for these cancer stem cells.