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目的:通过观察低氧对SGC7901细胞株SP细胞的影响及其与HIF-2α和ABCG2表达变化的关系,初步探讨低氧对肿瘤干细胞的影响及机制。方法:用氯化钴(CoCl2)创建低氧模型。Hoechst33342染色法检测SP细胞比例,免疫荧光细胞化学法检测ABCG2在SP和非SP细胞中的表达,免疫细胞化学法检测HIF-2α和ABCG2蛋白的表达,RT-PCR法检测ABCG2mRNA的表达。结果:SP细胞比例为2.36%,低氧诱导后其比例逐渐增加,P<0.05。ABCG2在SP细胞中的表达明显高于非SP细胞(P<0.01),低氧诱导后在SP细胞和非SP细胞中的表达均增强(P值均<0.01),且在SP细胞中的表达高于非SP细胞,P<0.01。低氧诱导后HIF-2α、ABCG2蛋白及ABCG2mRNA的表达较常氧组明显增强(P值均<0.01),且HIF-2α与ABCG2蛋白表达呈高度正相关(n=4,r=0.893,P<0.05)。结论:低氧能促进SGC7901细胞株SP细胞比例的增加,其机制可能与低氧激活HIF-2-αABCG2通路的表达,促进肿瘤细胞的干细胞化及维持干细胞的未分化状态有关。
OBJECTIVE: To investigate the effect of hypoxia on the proliferation of SGC7901 cell line SP cells and its relationship with the changes of HIF-2α and ABCG2 expression, and to explore the possible mechanism of hypoxia on tumor stem cells. Methods: An hypoxic model was created with cobalt chloride (CoCl2). The proportion of SP cells was detected by Hoechst33342 staining. The expression of ABCG2 in SP and non-SP cells was detected by immunofluorescence cytochemistry. The expression of HIF-2α and ABCG2 protein was detected by immunocytochemistry. The expression of ABCG2 mRNA was detected by RT-PCR. Results: The proportion of SP cells was 2.36%, and the proportion of SP cells increased gradually after hypoxia induction (P <0.05). The expression of ABCG2 in SP cells was significantly higher than that in non-SP cells (P <0.01). The expression of ABCG2 in SP cells was significantly increased after hypoxia induction (P <0.01), and the expression of ABCG2 in SP cells Higher than non-SP cells, P <0.01. The expression of HIF-2α, ABCG2 protein and ABCG2 mRNA in hypoxia group was significantly higher than that in normoxia group (all P <0.01), and the expression of HIF-2α and ABCG2 protein were highly correlated (P = 0.008, P = <0.05). CONCLUSION: Hypoxia can promote the increase of the proportion of SP cells in SGC7901 cells, which may be related to the hypoxic activation of HIF-2-αABCG2 pathway, the promotion of tumor cell stem cell and the maintenance of the undifferentiated state of stem cells.