CAJ | 학술논문

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.
목적 종인이선암세포계SW1990중분리종류간세포양측군(side population,SP)세포,병진일보연구기생물학특성.방법 세포상규배양후응용Hoechst 33342화PI염색、형광격발류식세포의검측병분선SP세포,MTT법검측세포활성,류식세포의분석간세포표지물CD133적표체,극륭평판측정세포극륭형성능력,Western blot검측세포ABC초가족성원G2(ABCG2)단백표체.결과 인이선암세포계SW1990중SP세포비례위2.70%,가완전피유랍파미조단.배양9 d후,SP세포적A492치위2.1,극륭형성솔위(38.7±6.8)%,CD133양성솔위69.63%,균현저고우비SP세포적0.5,(15.5±2.8)%,16.71%;SP세포ABCG2표체야교비SP세포강.결론 이선암세포계SW1990존재SP세포.
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.