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目的:探讨呼肠孤病毒对乳腺癌微球体细胞凋亡的影响。方法:以干细胞培养液培养乳腺癌细胞MCF-7和BT474,形成微球体,分别用呼肠孤病毒和表柔比星处理微球体,FCM检测MCF-7、BT474微球体细胞中CD44+CD24-/low细胞的比例,TUNEL法检测微球体细胞的凋亡,Western blotting检测MCF-7细胞及其微球体细胞中Ras的表达。结果:呼肠孤病毒对乳腺癌MCF-7和BT474微球体细胞的感染率显著高于亲本细胞(均P<0.05)。表柔比星作用下MCF-7和BT474微球体细胞中CD44+CD24-/low细胞比例明显升高[(8.1±0.4)%vs(18.0±4.5)%,P<0.01;(0.6±0.2)%vs(0.9±0.13)%,P<0.05]。呼肠孤病毒感染后MCF-7和BT474微球体细胞中CD44+CD24-/low细胞比例明显下降[(8.1±0.4)%vs(0.8±0.2)%,(0.6±0.2)%vs(0.2±0.1)%;均P<0.05]。MCF-7和BT474微球体细胞在呼肠孤病毒感染后凋亡率明显增加[(1.90±0.21)%vs(5.0±1.4)%,P<0.05;(0.20±0.1)%vs(1.0±0.16)%,P<0.05]。MCF-7细胞及其微球体细胞均高表达Ras,且两者水平相似。结论:与表柔比星相比,呼肠弧病毒对乳腺癌微球体细胞的抑制作用明显,还可明显诱导乳腺癌微球体细胞凋亡。
Objective: To investigate the effect of reovirus on apoptosis of breast cancer microspheres. METHODS: Breast cancer cells MCF-7 and BT474 were cultured in stem cell culture medium to form microspheres. Microspheres were treated with reovirus and epirubicin, respectively. FCM was used to detect CD44+CD24- in MCF-7 and BT474 microspheres. The proportion of /low cells was detected by TUNEL method. The expression of Ras in MCF-7 cells and microspheres was detected by Western blotting. Results: The infection rate of reovirus on MCF-7 and BT474 microsphere cells was significantly higher than that of parental cells (all P<0.05). The ratio of CD44+CD24-/low cells in MCF-7 and BT474 microspheres increased significantly with epirubicin [(8.1±0.4)% vs (18.0±4.5)%, P<0.01; (0.6±0.2) %vs (0.9±0.13)%, P<0.05]. The ratio of CD44+CD24-/low cells in MCF-7 and BT474 microspheres was significantly decreased after reovirus infection [(8.1±0.4)% vs (0.8±0.2)%, (0.6±0.2)% vs (0.2±) 0.1)%; all P<0.05]. The apoptosis rate of MCF-7 and BT474 microsphere cells increased significantly after reovirus infection [(1.90±0.21)% vs (5.0±1.4)%, P<0.05; (0.20±0.1)% vs (1.0±0.16) )%, P<0.05]. Ras was highly expressed in both MCF-7 cells and their microspheres, and the levels were similar. CONCLUSION: Compared with epirubicin, reovirus has a significant inhibitory effect on breast cancer microsphere cells, and can significantly induce breast cancer microsphere cell apoptosis.