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目的:观察耐吉西他滨胰腺癌细胞株(SW1990/GZ)上皮间质转化的现象。方法:倒置显微镜下观察亲代SW1990细胞与SW1990/GZ细胞的形态差别;流式细胞仪检测细胞周期;免疫印迹法(Western blot)检测上皮间质转化相关蛋白表达;Transwell小室测定细胞侵袭迁移能力;通过表面特异抗原(CD44+、CD24+和CD133+)标记流式细胞仪检测肿瘤干细胞比例。结果:在形态学上,SW1990/GZ表现出明显的间质细胞特征,与亲代SW1990相比处于G1期的细胞无明显差异;SW1990/GZ细胞低表达上皮细胞标志物E-cadherin,而高表达间质细胞的标记物Vimentin;SW1990/GZ细胞侵袭和迁移能力明显增强(P<0.01);与亲代细胞SW1990相比,SW1990/GZ细胞中CD44+CD24+细胞比例以及CD133+细胞比例分别提高了2倍和4倍以上(P<0.01)。结论:SW1990耐吉西他滨细胞株(SW1990/GZ)发生了明显的上皮间质转化过程,肿瘤干细胞比例明显增多。
Objective: To observe the phenomenon of epithelial-mesenchymal transition in the gemcitabine-resistant pancreatic cancer cell line (SW1990 / GZ). Methods: The morphology of SW1990 / SW1990 / GZ cells was observed under an inverted microscope. The cell cycle was detected by flow cytometry. The expression of epithelial-mesenchymal transition protein was detected by Western blot. The invasion and migration of SW1990 / Tumor stem cell ratios were measured by flow cytometry using surface-specific antigens (CD44 +, CD24 + and CD133 +). Results: Morphologically, SW1990 / GZ showed obvious characteristics of interstitial cells. There was no significant difference in G1 phase between SW1990 / GZ cells and SW1990 / GZ cells. However, SW1990 / GZ cells expressed low level of E-cadherin, Compared with SW1990, the percentage of CD44 + CD24 + cells and CD133 + cells in SW1990 / GZ cells were increased by 2 times compared with that of SW1990 / GZ cells (P <0.01) And 4 times more (P <0.01). CONCLUSIONS: SW1990 resistant gemcitabine cell line (SW1990 / GZ) undergoes obvious epithelial-mesenchymal transition and the proportion of tumor stem cells is significantly increased.