摘要
目的 探讨干细胞相关因子三磷酸腺苷（ATP）结合转运蛋白G超家族成员2（ATP-binding cassette transporter 2，ABCG2）在人皮肤鳞状细胞癌（鳞癌）组织及A431侧群细胞中的表达。方法 培养人A431细胞，流式细胞仪分选侧群细胞，噻唑蓝（MTT）实验比较侧群、非侧群细胞生长的增殖能力，逆转录PCR检测ABCG2在两者中的表达。免疫组化MaxVision法检测人鳞癌组织中ABCG2表达情况。结果 A431细胞系中存在侧群细胞，约占总细胞数的1.1%。侧群细胞具有较强的生长能力以及克隆形成能力，侧群组和非侧群组24孔板内每孔克隆形成数分别为114.8 ± 4.95和44.5 ± 3.67（t = 27.92，P ＜ 0.01），且侧群细胞ABCG2 mRNA表达量明显高于非侧群细胞（t = 5.22，P ＜ 0.01）。在人鳞癌组织中存在少量ABCG2阳性细胞，ABCG2蛋白主要表达在细胞胞质和胞膜上。结论 人鳞癌细胞A431中存在具有干细胞特性的侧群细胞，高表达ABCG2。ABCG2可作为鳞癌干细胞的表面标志物之一。
Objective To investigate the expression of stem cell marker adenosine triphosphate （ATP）-binding cassette transporter 2 （ABCG2） in the tissue and side population （SP） of a cell line A431 of human cutaneous squamous cell carcinoma （SCC）. Methods SP cells were separated by flow cytometry from cultured A431 cells. Methyl thiazolyl tetrazolium （MTT） assay was used to evaluate the proliferative ability of, reverse transcription-PCR to determine the expression of ABCG2 in, SP and non-SP cells. Immunohistochemistry （MaxVision method） was carried out to detect the expression of ABCG2 protein in tissue specimens from 10 patients with SCC. Results SP cells existed in cultured A431 cells, and accounted for about 1.1% of A431 cells. The SP cells had a stronger growth and colony-forming ability than non-SP cells did. The number of cell clones formed by SP cells and non-SP cells was 114.8 ± 4.95 and 44.5 ± 3.67, respectively, per well in a 24-well plate （t = 27.92, P 〈 0.01）. The expression level of ABCG2 mRNA was significantly higher in SP cells than in non-SP cells （t = 5.22, P 〈 0.01）. There existed a small number of ABCG2 positive cells in SCC tissue, and ABCG2 was mainly expressed in the cytoplasm and membrane of tumor cells. Conclusions SP cells exist in A431 cells, which have characteristics of stem cells and highly express ABCG2. ABCG2 may be a potential stem cell surface marker of SCC.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2012年第6期392-395,共4页
Chinese Journal of Dermatology
基金
基金项目：江苏省自然科学基金（BK2009063）
