人胰腺癌细胞系SW1990中肿瘤干细胞样侧群细胞的分离及生物学特性研究

目的 从人胰腺癌细胞系SW1990中分离肿瘤干细胞样侧群(side population,SP)细胞,并进一步研究其生物学特性.方法 细胞常规培养后应用Hoechst 33342和PI染色、荧光激发流式细胞仪检测并分选SP细胞,MTT法检测细胞活性,流式细胞仪分析干细胞标志物CD133的表达,克隆平板测定细胞克隆形成能力,Western blot检测细胞ABC超家族成员G2(ABCG2)蛋白表达.结果 人胰腺癌细胞系SW1990中SP细胞比例为2.70%,可完全被维拉帕米阻断.培养9 d后,SP细胞的A492值为2.1,克隆形成率为(38.7±6.8)%,CD133阳性率为69.63%,均显著高于非SP细胞的0.5,(15.5±2.8)%,16.71%;SP细胞ABCG2表达也较非SP细胞强.结论 胰腺癌细胞系SW1990存在SP细胞.

Identification of side population cells like cancer stem cell in human pancreatic cancer cell line SW1990

Objective To isolate and identify side population (SP) cells like cancer stem cell from human pancreatic cancer cell line SW1990, for the purpose of further evaluation of their biological characteristics. Methods Cell suspension was stained with Hoechst 33342 and PI. Then SP cells were analyzed in the fluorescence activated cell sorter. Cell growth viability was measured by MTT. Stem cell marker CD133 was determined by flow cytometry. Cloning forming efficiency was determined by cloning plating. Expression of ABCG2 protein was detected by Western blot analysis. Results The proportion of SP cells was 2.7%, however it could be completely blocked by verapamil. 9 days later, the value of A492 of SP cells was 2.1, the cloning forming efficiency was (38.7 ± 6.8) % , the positive rate of CD133 was 69.63%, which were significantly higher than cells 0. 5, ( 15.5 ± 2.8)%, 16.71% of corresponding non-SP( P <0.05). The expression of ABCG2 in SP cells was significantly higher than that in non-SP cells. Conclusions SP cells existed in human pancreatic cancer cells SW1990.