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目的:根据肿瘤干细胞学说理论,探讨胰腺癌耐药的新机制。方法:通过胰腺癌肿瘤干细胞表面标记途径和侧群(side population,SP)细胞途径,分离出人胰腺癌细胞株PANC-1中的SP/NSP(非SP)细胞及CD44+CD24+/CD44-CD24-细胞亚群,用MTT检测上述各亚群细胞在体外对化疗药物耐受的差异,用AnnexinV-PI双染法检测2种肿瘤细胞的抗凋亡能力,并采用实时荧光定量PCR检测两者耐药基因ABCG2、ABCB1和PLK-1表达的差异。结果:人胰腺癌细胞株PANC-1中的SP细胞比例为(7.64±0.96)%,CD44+CD24+细胞比例为(2.60±0.96)%。相对于NSP和CD44-CD24-细胞而言,SP和CD44+CD24+细胞具有更强的化疗耐受能力(P<0.01)和抗凋亡能力(P<0.01);荧光定量RT-PCR结果均提示,SP和CD44+CD24+细胞高表达耐药基因ABCG2、ABCB1和PLK1。结论:人胰腺癌细胞株PANC-1中SP及CD44+CD24+细胞具有更强的化疗耐受能力,研究胰腺癌肿瘤干细胞可为克服胰腺癌化疗敏感性差的现状提供新的实验基础和理论依据。
OBJECTIVE: To explore a new mechanism of drug resistance in pancreatic cancer based on the theory of tumor stem cell theory. METHODS: SP / NSP (non-SP) cells and CD44 + CD24 + / CD44-CD24 in human pancreatic cancer cell line PANC-1 were isolated by pancreatic cancer stem cell surface marker and side population (SP) - cell subsets. The tolerance of chemotherapeutic drugs to these subpopulations in vitro was assayed by MTT assay. The anti-apoptotic ability of the two tumor cells was detected by Annexin V-PI double staining and real-time fluorescent quantitative PCR Differences in the expression of drug resistance genes ABCG2, ABCB1 and PLK-1. Results: The percentage of SP cells in human pancreatic cancer cell line PANC-1 was (7.64 ± 0.96)% and that of CD44 + CD24 + cells was (2.60 ± 0.96)%. Compared with NSP and CD44-CD24- cells, SP and CD44 + CD24 + cells had stronger chemoresistance (P <0.01) and anti-apoptosis ability (P <0.01) , And high expression of resistance genes ABCG2, ABCB1 and PLK1 in SP and CD44 + CD24 + cells. Conclusions: SP and CD44 + CD24 + cells in human pancreatic cancer cell line PANC-1 have stronger chemoresistance. To study pancreatic cancer stem cells may provide a new experimental basis and theoretical basis for overcoming the poor sensitivity of pancreatic cancer chemotherapy.