中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2012年
6期
392-395
,共4页
李泓馨%赵勇%管海宏%林麟
李泓馨%趙勇%管海宏%林麟
리홍형%조용%관해굉%림린
A431细胞%癌,鳞状细胞%肿瘤干细胞%ABCG2%侧群细胞
A431細胞%癌,鱗狀細胞%腫瘤榦細胞%ABCG2%側群細胞
A431세포%암,린상세포%종류간세포%ABCG2%측군세포
A431 cells%Carcinoma,squamous cell%Tumor stem cells%ABCG2%Side population cells
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.
目的 探討榦細胞相關因子三燐痠腺苷(ATP)結閤轉運蛋白G超傢族成員2(ATP-binding cassette transporter 2,ABCG2)在人皮膚鱗狀細胞癌(鱗癌)組織及A431側群細胞中的錶達.方法 培養人A431細胞,流式細胞儀分選側群細胞,噻唑藍(MTT)實驗比較側群、非側群細胞生長的增殖能力,逆轉錄PCR檢測ABCG2在兩者中的錶達.免疫組化MaxVision法檢測人鱗癌組織中ABCG2錶達情況.結果 A431細胞繫中存在側群細胞,約佔總細胞數的1.1％.側群細胞具有較彊的生長能力以及剋隆形成能力,側群組和非側群組24孔闆內每孔剋隆形成數分彆為114.8±4.95和44.5±3.67(t=27.92,P＜0.01),且側群細胞ABCG2 mRNA錶達量明顯高于非側群細胞(t=5.22,P＜ 0.01).在人鱗癌組織中存在少量ABCG2暘性細胞,ABCG2蛋白主要錶達在細胞胞質和胞膜上.結論 人鱗癌細胞A431中存在具有榦細胞特性的側群細胞,高錶達ABCG2.ABCG2可作為鱗癌榦細胞的錶麵標誌物之一.
목적 탐토간세포상관인자삼린산선감(ATP)결합전운단백G초가족성원2(ATP-binding cassette transporter 2,ABCG2)재인피부린상세포암(린암)조직급A431측군세포중적표체.방법 배양인A431세포,류식세포의분선측군세포,새서람(MTT)실험비교측군、비측군세포생장적증식능력,역전록PCR검측ABCG2재량자중적표체.면역조화MaxVision법검측인린암조직중ABCG2표체정황.결과 A431세포계중존재측군세포,약점총세포수적1.1％.측군세포구유교강적생장능력이급극륭형성능력,측군조화비측군조24공판내매공극륭형성수분별위114.8±4.95화44.5±3.67(t=27.92,P＜0.01),차측군세포ABCG2 mRNA표체량명현고우비측군세포(t=5.22,P＜ 0.01).재인린암조직중존재소량ABCG2양성세포,ABCG2단백주요표체재세포포질화포막상.결론 인린암세포A431중존재구유간세포특성적측군세포,고표체ABCG2.ABCG2가작위린암간세포적표면표지물지일.
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.
