Effect of siRNA-silencing of SALL2 gene on growth, migration and invasion of human ovarian carcinoma A2780 cells.
Effect of siRNA-silencing of SALL2 gene on growth, migration and invasion of human ovarian carcinoma A2780 cells.
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DOI：
10.1186/s12885-017-3843-y
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发表时间：
2017-12-11
期刊：
影响因子：
3.8
通讯作者：
Zhang X
中科院分区：
文献类型：
--
作者：
Miao F;Zhang X;Cao Y;Wang Y;Zhang X
Spalt-like gene-2(SALL2)在肿瘤发生中的作用尚未完全阐明。本研究探讨SALL2对人卵巢癌A2780细胞的作用及其可能机制。采用逆转录聚合酶链式反应(RT-PCR)和Western印迹分析检测SALL2在人卵巢癌细胞系中的表达。将小干扰核糖核酸(SiRNA)导入A2780细胞，使SALL2沉默。RT-PCR、Western印迹分析和免疫荧光检测SALL2的表达。用CCK-8比色法和流式细胞仪检测细胞增殖。用流式细胞仪检测细胞凋亡率。实时细胞分析检测细胞迁移情况。Transwell法检测细胞侵袭力。实时荧光定量聚合酶链式反应检测p21基因的表达。免疫印迹法检测基质金属蛋白酶(MMP2)、MMP9、蛋白激酶B(PKB，又称Akt)和磷酸化Akt(p-Akt)的表达。SALL2在6个OC细胞系中均有表达，其中A2780细胞的表达水平最高。与siRNA-2和siRNA-3组相比，siRNA-2和siRNA-3转染48 h后，A2780细胞SALL2表达下调，24、48和72 h后，siRNA-2组A2780细胞增殖增强，A2780细胞处于G0/G1期比例下降，48 h后细胞凋亡率降低，细胞迁移和侵袭能力增强。与干扰组相比，siRNA2转染后A2780细胞中p21mRNA的表达降低。当SALL2被沉默时，A2780细胞MMP2/9和p-Akt的表达增加。此外，PI3K抑制剂LY294002可有效逆转SALL2 siRNA诱导的Akt的磷酸化、A2780细胞的迁移和侵袭。瞬时沉默SALL2可促进细胞增殖、迁移和侵袭，抑制A2780细胞的凋亡。在SALL2 siRNA沉默的细胞中，p21的表达降低。SiRNA下调SALL2基因可诱导A2780细胞的迁移和侵袭，这一现象可能与MMP2/9表达增加和PI3K/Akt信号通路激活有关。本文的在线版本(10.1186/s12885-0173843-y)包含向授权用户提供的补充材料。
The role of Spalt-like gene-2 (SALL2) in tumorigenesis remains incompletely elucidated. This study investigated the effects of SALL2 on human ovarian carcinoma (OC) A2780 cells and the probable mechanism. Expression of SALL2 in human OC cell lines were detected by reverse transcription PCR (RT-PCR) and Western blot analysis. A2780 cells were transfected with small-interfering ribonucleic acid (siRNA) to silence SALL2. SALL2 expression was detected by RT-PCR, Western blot analysis and immunofluorescence assay. Cell proliferation was measured by CCK-8 assay and flow cytometry (FCM). Apoptosis was measured by FCM. Cell migration was detected by real-time cell analysis. Cell invasion was detected by transwell assay. mRNA expression of p21 was detected by quantitative real-time PCR. Western blot analysis was used to determine the expression of matrix metalloproteinase (MMP)2, MMP9, protein kinase B (PKB, also called Akt), and phosphorylated-Akt (p-Akt). SALL2 was expressed in six OC cell lines, and the expression was the highest in A2780 cells. Compared with that in the Scramble group, SALL2 expression in A2780 was downregulated after transfection with siRNA-2 and siRNA-3 for 48 h. Compared with that in the Scramble group, proliferation of A2780 cells in the siRNA-2 group increased after transfection for 24, 48 and 72 h. In the siRNA-2 group, the proportion of A2780 cells decreased in the G0/G1 phase, and cell apoptosis decreased after transfection for 48 h. Compared with that in the Scramble group, the cell migration and invasion abilities of A2780 cells increased. Compared with that in the Scramble group, p21 mRNA expression in A2780 cells decreased after transfection with siRNA2. When SALL2 was silenced, the expression of MMP2/9 and p-Akt in A2780 cells increased. Furthermore, the PI3K inhibitor LY294002 could effectively reversed SALL2 siRNA-induced phosphorylation of Akt, migration and invasion of A2780 cells. Transient silencing of SALL2 promotes cell proliferation, migration, and invasion, and inhibits apoptosis of A2780 cells. In SALL2 siRNA-silenced cells, p21 expression was decreased. SALL2 knockdown by siRNA induces the migration and invasion of A2780 cells; this phenomenon is possibly associated with the increased expression of MMP2/9 and the activation of the PI3K/Akt signalling pathway. The online version of this article (10.1186/s12885-017-3843-y) contains supplementary material, which is available to authorized users.
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DOI：
10.1016/j.bbagrm.2011.02.002
发表时间：
2011-04
影响因子：
4.7
作者：
Gu, Hongcang;Li, Dawei;Sung, Chang K.;Yim, Hyungshin;Troke, Philip;Benjamin, Thomas
通讯作者：
Benjamin, Thomas
影响因子：
2.6
作者：
Buck, A;Kispert, A;Kohlhase, J
通讯作者：
Kohlhase, J
影响因子：
5.3
作者：
Parrish, M;Ott, T;Monaghan, AP
通讯作者：
Monaghan, AP
影响因子：
3.6
作者：
Jin, Yunfeng;Huang, Menghui;Xi, Qinghua
通讯作者：
Xi, Qinghua
影响因子：
2.8
作者：
Baulch, Janet E.;Geidzinski, Erich;Limoli, Charles L.
通讯作者：
Limoli, Charles L.