Sinomenine Inhibits the Growth of Ovarian Cancer Cells Through the Suppression of Mitosis by Down-Regulating the Expression and the Activity of CDK1.
Sinomenine Inhibits the Growth of Ovarian Cancer Cells Through the Suppression of Mitosis by Down-Regulating the Expression and the Activity of CDK1.
复制标题
青藤碱通过下调 CDK1 的表达和活性来抑制有丝分裂，从而抑制卵巢癌细胞的生长。
DOI：
10.2147/ott.s284261
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发表时间：
2021
影响因子：
4
通讯作者：
Cheng Z
中科院分区：
文献类型：
--
作者：
Qu X;Yu B;Zhu M;Li X;Ma L;Liu C;Zhang Y;Cheng Z
卵巢癌是世界上最常见的妇科癌症之一。然而，针对卵巢癌的治疗尚未完全有效，青藤碱已被证明在多种癌细胞中具有抗肿瘤活性。但其抗卵巢癌作用的研究尚不多见，其机制尚未阐明。因此，本研究旨在探讨青藤碱抗卵巢癌的作用机制。CCK-8法和集落形成实验检测其抗卵巢癌HeyA 8细胞的作用。采用高通量RNA-seq技术探讨青藤碱抗卵巢癌的作用机制，并分别采用实时荧光定量PCR和Western blot技术验证靶mRNA和蛋白的表达。我们发现，卵巢癌HeyA 8细胞的增殖和克隆形成能力显着降低1.56 mM青藤碱。转录组分析显示，青藤碱处理后HeyA 8细胞中有2679个基因发生了差异表达，其中下调基因1323个，上调基因1356个。基因本体和KEGG途径富集表明，青藤碱和DMSO处理的HeyA 8细胞组间差异表达基因（DEG）主要参与细胞周期过程，如着丝点组装、染色体分离和DNA复制。值得注意的是，在蛋白质-蛋白质相互作用（PPI）网络中排名前18位的基因主要参与有丝分裂过程，如姐妹染色单体分离，染色体浓缩和微管细胞骨架组织。此外，实时PCR结果显示与转录组分析一致的DEG表达趋势。Western blot结果显示，青藤碱处理后，PPI中最高级的基因、控制有丝分裂过程的主要调控因子CDK 1的表达水平降低，磷酸化的P-CDK（Thr 161）和P-Histone H3（Ser 10）水平降低。结果表明青藤碱通过下调CDK 1的表达和活性抑制HeyA 8细胞的有丝分裂，从而抑制HeyA 8细胞的增殖。本研究为青藤碱抗卵巢癌的应用提供了初步的研究依据。
Ovarian cancer is one of the most common gynecological cancers worldwide. While, therapies against ovarian cancer have not been completely effective, sinomenine has been proved to have anti-tumor activity in various cancer cells. However, study of its anti-ovarian cancer effect is still rare, and the underlying mechanism has not been elucidated. Therefore, we aim to explore the mechanism of sinomenine anti-ovarian cancer. The effect of anti-ovarian cancer HeyA8 cells was analyzed by CCK8 and colony formation assay. The mechanism of sinomenine anti-ovarian cancer was explored via high throughput RNA-seq, and then the target mRNA and protein expression were verified by real-time PCR and Western blot, respectively. We found that the proliferation and clone formation ability of ovarian cancer HeyA8 cells were markedly reduced by 1.56 mM sinomenine. The transcriptome analysis showed that 2679 genes were differentially expressed after sinomenine treatment in HeyA8 cells, including 1323 down-regulated genes and 1356 up-regulated genes. Gene ontology and KEGG pathway enrichment indicated that differential expression genes (DEGs) between the groups of sinomenine and DMSO-treated HeyA8 cells were mainly involved in the process of the cell cycle, such as kinetochore organization, chromosome segregation, and DNA replication. Strikingly, the top 18 ranked degree genes in the protein–protein interaction (PPI) network were mainly involved in the process of mitosis, such as sister chromatid segregation, condensed chromosome, and microtubule cytoskeleton organization. Moreover, real-time PCR results showed consistent expression trends of DEGs with transcriptome analysis. The results of Western blot showed the expression level of CDK1, which was the highest degree gene in PPI and the main regulator controlling the process of mitosis, and the levels of phosphorylated P-CDK (Thr161) and P-Histone H3 (Ser10) were decreased after being treated with sinomenine. Our results demonstrated that sinomenine inhibited the proliferation of HeyA8 cells through suppressing mitosis by down-regulating the expression and the activity of CDK1. The study may provide a preliminary research basis for the application of sinomenine in anti-ovarian cancer.