Isolation, cultivation and identification of human lung adenocarcinoma stem cells

Zhang,De‑Geng; Jiang,Ai‑Gui; Lu,Hui‑Yu; Zhang,Li‑Xin; Gao,Xiao‑Yan

doi:10.3892/ol.2014.2639

Isolation, cultivation and identification of human lung adenocarcinoma stem cells

Authors:
- De‑Geng Zhang
- Ai‑Gui Jiang
- Hui‑Yu Lu
- Li‑Xin Zhang
- Xiao‑Yan Gao
View Affiliations

Affiliations: Department of Oncology, Taizhou People's Hospital Affiliated to Medical College of Nantong University, Taizhou, Jiangsu 225300, P.R. China, Department of Respiratory Medicine, Taizhou People's Hospital Affiliated to Medical College of Nantong University, Taizhou, Jiangsu 225300, P.R. China, Institute of Clinical Medicine, Taizhou People's Hospital Affiliated to Medical College of Nantong University, Taizhou, Jiangsu 225300, P.R. China
Published online on: October 24, 2014 https://doi.org/10.3892/ol.2014.2639
Pages: 47-54

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Recently, an increasing number of studies have demonstrated that lung cancer is a stem cell disease. However, ideal cell surface markers for isolating stem cells in lung cancer are yet to be identified. In the present study, a cell population with a cluster of differentiation (CD)133+ phenotype was successfully isolated from a single cell suspension of lung adenocarcinoma tissue using magnetic‑activated cell sorting (MACS) and enriched in a serum‑free culture. In comparison to CD133‑ cells, the CD133+ cells exhibited an enhanced capacity for self‑renewal and differentiation, and a greater potential for in vivo tumor formation, in non‑obese diabetic/severe combined immunodeficient (NOD/SCID) mice. Tumors could be induced in NOD/SCID mice by the transplantation of 102 stem‑like cells per mouse. The results of the present study demonstrated that CD133 may serve as a specific cell surface marker for lung adenocarcinoma stem cells, and that MACS combined with serum‑free culture is an effective method for isolating and enriching lung cancer stem cells.

View Figures

View References

1	Molina JR, Yang P, Cassivi SD, Schild SE and Adjei AA: Non-small cell lung cancer: epidemiology, risk factors, treatment, and survivorship. Mayo Clin Proc. 83:584–594. 2008. View Article : Google Scholar : PubMed/NCBI
2	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar : PubMed/NCBI
3	Yang L, Parkin DM, Ferlay J, Li L and Chen Y: Estimates of cancer incidence in China for 2000 and projections for 2005. Cancer Epidemiol Biomarkers Prev. 14:243–250. 2005.PubMed/NCBI
4	Zhou XD, Wang XY, Qu FJ, et al: Detection of cancer stem cells from the C6 glioma cell line. J Int Med Res. 37:503–510. 2009. View Article : Google Scholar : PubMed/NCBI
5	Qiu B, Zhang D, Tao J, Wu A and Wang Y: A simplified and modified procedure to culture brain glioma stem cells from clinical specimens. Oncol Lett. 3:50–54. 2012.PubMed/NCBI
6	Hwang-Verslues WW, Kuo WH, Chang PH, et al: Multiple lineages of human breast cancer stem/progenitor cells identified by profiling with stem cell markers. PLoS One. 4:e83772009. View Article : Google Scholar : PubMed/NCBI
7	Chen W and Wang GM: Gene expression profiling of cancer stem cells in the Du145 prostate cancer cell line. Oncol Lett. 3:791–796. 2012.PubMed/NCBI
8	O’Brien CA, Pollett A, Gallinger S and Dick JE: A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature. 445:106–110. 2007. View Article : Google Scholar
9	Suetsugu A, Nagaki M, Aoki H, Motohashi T, Kunisada T and Moriwaki H: Characterization of CD133⁺ hepatocellular carcinoma cells as cancer stem/progenitor cells. Biochem Biophys Res Commun. 351:820–824. 2006. View Article : Google Scholar : PubMed/NCBI
10	Varnat F, Duquet A, Malerba M, et al: Human colon cancer epithelial cells harbour active HEDGEHOG-GLI signalling that is essential for tumour growth, recurrence, metastasis and stem cell survival and expansion. EMBO Mol Med. 1:338–351. 2009. View Article : Google Scholar
11	Reya T, Morrison SJ, Clarke MF and Weissman IL: Stem cells, cancer and cancer stem cells. Nature. 414:105–111. 2001. View Article : Google Scholar : PubMed/NCBI
12	Sullivan JP, Minna JD and Shay JW: Evidence for self-renewing lung cancer stem cells and their implications in tumour initiation, progression, and targeted therapy. Cancer Metastasis Rev. 29:61–72. 2010. View Article : Google Scholar : PubMed/NCBI
13	Otto WR: Lung epithelial stem cells. J Pathol. 197:527–535. 2002. View Article : Google Scholar : PubMed/NCBI
14	Rawlins EL and Hogan BL: Epithelial stem cells of the lung: privileged few or opportunities for many? Development. 133:2455–2465. 2006. View Article : Google Scholar : PubMed/NCBI
15	Shi Y, Fu X, Hua Y, Han Y, Lu Y and Wang J: The side population in human lung cancer cell line NCI-H460 is enriched in stem-like cancer cells. PLoS One. 7:e333582012. View Article : Google Scholar : PubMed/NCBI
16	Meng M, Zhao XH, Ning Q, Hou L, Xin GH and Liu LF: Tumor stem cells: A new approach for tumor therapy (Review). Oncol Lett. 4:187–193. 2012.PubMed/NCBI
17	Kim CF, Jackson EL, Woolfenden AE, et al: Identification of bronchioalveolar stem cells in normal lung and lung cancer. Cell. 121:823–835. 2005. View Article : Google Scholar : PubMed/NCBI
18	Ho MM, Ng AV, Lam S and Hung JY: Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells. Cancer Res. 67:4827–4833. 2007. View Article : Google Scholar : PubMed/NCBI
19	Eramo A, Lotti F, Sette G, et al: Identification and expansion of the tumorigenic lung cancer stem cell population. Cell Death Differ. 15:504–514. 2008. View Article : Google Scholar
20	Groome PA, Bolejack V, Crowley JJ, et al: IASLC International Staging Committee; Cancer Research and Biostatistics; Observers to the Committee; Participating Institutions: The IASLC Lung Cancer Staging Project: validation of the proposals for revision of the T, N, and M descriptors and consequent stage groupings in the forthcoming (seventh) edition of the TNM clas-sification of malignant tumors. J Thorac Oncol. 2:694–705. 2007. View Article : Google Scholar : PubMed/NCBI
21	Dong QG, Yao M, Geng Q, Zhou J and Yan MX: Isolation and identification of human lung adenocarcinoma stem cells. Tumor. 28:1–7. 2008.(In Chinese).
22	Bertolini G, Roz L, Perego P, et al: Highly tumorigenic lung cancer CD133+ cells display stem-like features and are spared by cisplatin treatment. Proc Natl Acad Sci USA. 106:16281–16286. 2009. View Article : Google Scholar : PubMed/NCBI
23	Dean M: ABC transporters, drug resistance, and cancer stem cells. J Mammary Gland Biol Neoplasia. 14:3–9. 2009. View Article : Google Scholar : PubMed/NCBI
24	Sung JM, Cho HJ, Yi H, et al: Characterization of stem cell population in lung cancer A549 cells. Biochem Biophys Res Commun. 371:163–167. 2008. View Article : Google Scholar : PubMed/NCBI
25	Levina V, Marrangoni AM, DeMarco R, Gorelik E and Lokshin AE: Drug-selected human lung cancer stem cells: cytokine network, tumorigenic and metastatic properties. PLoS One. 3:e30772008. View Article : Google Scholar : PubMed/NCBI
26	Omary MB, Coulombe PA and McLean WH: Intermediate filament proteins and their associated diseases. New Engl J Med. 351:2087–2100. 2004. View Article : Google Scholar : PubMed/NCBI
27	Fukunaga Y, Bandoh S, Fujita J, et al: Expression of cytokeratin 8 in lung cancer cell lines and measurement of serum cytokeratin 8 in lung cancer patients. Lung Cancer. 38:31–38. 2002. View Article : Google Scholar : PubMed/NCBI
28	Kanaji N, Bandoh S, Fujita J, Ishii T, Ishida T and Kubo A: Compensation of type I and type II cytokeratin pools in lung cancer. Lung Cancer. 55:295–302. 2007. View Article : Google Scholar
29	Ghotra VP, Puigvert JC and Danen EH: The cancer stem cell microenvironment and anti-cancer therapy. Int J Radiat Biol. 85:955–962. 2009. View Article : Google Scholar : PubMed/NCBI