[1]张彦,郑晓东,唐鹏,等.Sphk1基因对人乳腺癌MCF-7细胞增殖、凋亡和迁移能力的影响[J].第三军医大学学报,2012,34(21):2141-2144.
 Zhang Yan,Zheng Xiaodong,Tang Peng,et al.Sphk1 interference suppresses proliferation, apoptosis and migration in human MCF-7 breast cancer cells[J].J Third Mil Med Univ,2012,34(21):2141-2144.
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Sphk1基因对人乳腺癌MCF-7细胞增殖、凋亡和迁移能力的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第21期
页码:
2141-2144
栏目:
论著
出版日期:
2012-11-15

文章信息/Info

Title:
Sphk1 interference suppresses proliferation, apoptosis and migration in human MCF-7 breast cancer cells
作者:
张彦郑晓东唐鹏张毅姜军
第三军医大学西南医院乳腺中心
Author(s):
Zhang Yan Zheng Xiaodong Tang Peng Zhang Yi Jiang Jun
Center of Breast Disease, Southwest Hospital,Third Military Medical University,Chongqing, 400038, China
关键词:
乳腺肿瘤RNA干扰siRNASphk1
Keywords:
breast cancer RNA interference siRNA Sphk1
分类号:
R394.2;R730.23;R737.9
文献标志码:
A
摘要:
目的      通过抑制鞘氨醇激酶-1(sphingosine kinase-1,Sphk1)基因表达,观察其对人乳腺癌MCF-7细胞增殖、凋亡和迁移能力的影响。      方法       使用脂质体2000将化学合成的靶向Sphk1基因的小干扰RNA(small interfering RNA,siRNA)转入人乳腺癌MCF-7细胞。使用流式细胞术(FCM)检测转染效率,荧光定量RT-PCR(qRT-PCR)和 Western blot法检测Sphk1的mRNA和蛋白表达水平。利用CCK-8法检测细胞的增殖能力,流式细胞术检测细胞凋亡,Transwell小室法检测细胞迁移能力。      结果      Sphk1 siRNA转染效率达89%。转染siRNA后,MCF-7细胞的Sphk1 mRNA和蛋白表达水平均明显下降,以siRNA-2效果最为显著(P<0.01)。转染Sphk1 siRNA-2后,MCF-7细胞的增殖和迁移能力明显下降,凋亡细胞比例增加,以上差异均具有统计学意义(P<0.05)。      结论       干扰Sphk1基因表达可有效抑制MCF-7细胞的增殖、迁移能力,同时诱导细胞发生凋亡,Sphk1基因可能成为一个潜在的乳腺癌治疗靶标。
Abstract:
Objective      To investigate the effect of down-regulating sphingosine kinase-1 (Sphk1) on the proliferation, apoptosis and migration in human breast cancer MCF-7 cell line.       Methods      The siRNA against Sphk1 was chemically synthesized and then transfected into MCF-7 cells using Lipofectamine 2000. Transfection efficiency was examined by flow cytometry(FCM)and the expression levels of Sphk1 mRNA and protein were determined by real-time fluorescent quantitative PCR and Western blotting. Cell proliferation and cell apoptosis was assessed by CCK-8 assay and FCM respectively. Cell migration was measured by transwell assay.       Results      The efficiency of siRNA transfection into human MCF-7 cells were 89%. The siRNA against sphk1 gene significantly reduced the mRNA and protein expression of Sphk1 in MCF-7 cells (P<0.05). The siRNA-2 showed most significant effect(P<0.01).The MCF-7 cells transfected with Sphk1 siRNA-2 showed a significant decrease of cell proliferation and migration compared to the control cells. Moreover, significant increase of proportion of apoptotic cells were detected in the transfected cells(P<0.05).       Conclusion      Sphk1 knockdown in breast cancer cells effectively inhibits the proliferation and migration and induces the apoptosis in MCF-7 cells. Sphk1 may become a potential target for gene therapy of breast cancer.

参考文献/References:

张彦, 郑晓东, 唐鹏, 等. Sphk1基因对人乳腺癌MCF-7细胞增殖、凋亡和迁移能力的影响[J]. 第三军医大学学报,2012,34(21):2141-2144.

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更新日期/Last Update: 2012-11-06