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Comparative Study
. 2012 Sep 7;18(33):4610-7.
doi: 10.3748/wjg.v18.i33.4610.

Side population cells isolated from KATO III human gastric cancer cell line have cancer stem cell-like characteristics

Jun-Jun She  1 , Peng-Ge Zhang, Xuan Wang, Xiang-Ming Che, Zi-Ming Wang
Affiliations
Comparative Study

Side population cells isolated from KATO III human gastric cancer cell line have cancer stem cell-like characteristics

Jun-Jun She et al. World J Gastroenterol. .

Abstract

Aim: To investigate whether the side population (SP) cells possess cancer stem cell-like characteristics in vitro and the role of SP cells in tumorigenic process in gastric cancer.

Methods: We analyzed the presence of SP cells in different human gastric carcinoma cell lines, and then isolated and identified the SP cells from the KATO III human gastric cancer cell line by flow cytometry. The clonogenic ability and self-renewal were evaluated by clone and sphere formation assays. The related genes were determined by reverse transcription polymerase chain reaction. To compare tumorigenic ability, SP and non-side population (NSP) cells from the KATO III human gastric cancer cell line were subcutaneously injected into nude mice.

Results: SP cells from the total population accounted for 0.57% in KATO III, 1.04% in Hs-746T, and 0.02% in AGS (CRL-1739). SP cells could grow clonally and have self-renewal capability in conditioned media. The expression of ABCG2, MDRI, Bmi-1 and Oct-4 was different between SP and NSP cells. However, there was no apparent difference between SP and NSP cells when they were injected into nude mice.

Conclusion: SP cells have some cancer stem cell-like characteristics in vitro and can be used for studying the tumorigenic process in gastric cancer.

Keywords: Cancer stem cells; Gastric cancer; KATO III; Self-renewal; Side population.

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Figures

Figure 1
Figure 1
Side population and non-side population cells from several human gastric cancer cell lines were analyzed through uptake of the DNA binding dye Hoechst33342 with or without the presence of verapamil. NSP: Non-side population; SP: Side population.
Figure 2
Figure 2
Soft agar assays and fluorescence for KATO III-green fluorescent protein cells. A: Side population (SP) cells (A-2) were more clonogenic than non-side population (NSP) cells (A-1); B: The assay was repeated 3 times, and the column diagram indicated that there was a statistically significant difference in clonogenic efficiency between SP and NSP cells (aP < 0.05 vs NSP cells); C: KATO III-green fluorescent protein (GFP) cells expressed strong green fluorescence in regular culture, with no mixed colonies resulting from a single KATO III-GFP cell. C-1 was observed under white light, while C-2 was observed under fluorescent light.
Figure 3
Figure 3
Serial sphere assays. A: The assay was repeated 3 times, and the column diagram indicated a statistically significant difference in clonogenic efficiency between non-side population (NSP) and side population (SP) cells in primary and secondary sphere assays (aP < 0.05 vs NSP cells); however, there was no significant difference between primary and secondary sphere assays; B: Sphere assay for NSP cells; C: Sphere assay for SP cells.
Figure 4
Figure 4
Glyceraldehyde-3-phosphate dehydrogenase, adenosine-triphosphate-binding cassette sub-family G member 2, multidrug resistance protein 1, B-cell-specific Moloney murine leukemia virus insertion site 1 and octamer-binding transcription factor 4 RNA expression of side population and non-side population cells isolated from KATO III by reverse transcription polymerase chain reaction. SP: Side population; NSP: Non-side population; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; ABCG2: Adenosine-triphosphate-binding cassette sub-family G member 2; MDR1: Multidrug resistance protein 1; Bmi-1: B-cell-specific Moloney murine leukemia virus insertion site 1; Oct-4: Octamer-binding transcription factor 4.
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