[Isolation and identification of cancer stem cells from human esophageal carcinoma].

OBJECTIVE

To isolate and identify cancer stem cells from esophageal carcinoma cells (ECCs) using cell surface marker p75NTR.

METHODS

ECCs cell lines were established with ECCs collected from 38 surgically resected specimens. Flow cytometry was used to identify the p75NTR positive cells therein that were isolated then using magnetic activated cell sorting (MACS) method. The growing characteristics in DMEM medium and capability of colony-forming in soft agar of the p75NTR positive cells were evaluated ex vivo with MTT method. p75NTR positive cells of different concentrations were subcutaneously injected into the backs of Balb/c nude mice and PBS was injected into the contralateral back, and then tumorigenesis was observed, 8 weeks later the mice were killed with their tumors taken out to undergo microscopy.

RESULTS

Eight ECCs cell lines were established, 6 of which were found to contain 0.32%-3.35% of p75NTR positive cells. The purity of p75NTR positive cells isolated by MACS was up to 90%. MTT result showed that population doubling time of the p75NTR positive cells was (17+/-3) hours, significantly shorter than that of the p75NTR negative cells [(37+/-7) hours, P<0.01]. The colony-forming rate in soft agar of the p75NTR positive cells was (45.9%+/-8.9%), significantly higher than that of the p75NTR negative cells [(3.7%+/-2.1%), P<0.01]. As few as 2000 p75NTR positive cells gave rise to new tumors in xenotransplantation, with a tumorigenic ability 50 times as high as that of the p75NTR negative cells.

CONCLUSION

p75NTR positive cells carry some properties of cancer stem cells, such as the ability of self-renewal, differentiation and proliferation and demonstrate higher ability of colony-forming ex vivo and tumorigenesis in vivo.