Novel derivative of aminobenzenesulfonamide (3c) induces apoptosis in colorectal cancer cells through ROS generation and inhibits cell migration.
Novel derivative of aminobenzenesulfonamide (3c) induces apoptosis in colorectal cancer cells through ROS generation and inhibits cell migration.
复制标题
DOI：
10.1186/s12885-016-3005-7
复制
发表时间：
2017-01-03
期刊：
影响因子：
3.8
通讯作者：
Ahmad R
中科院分区：
文献类型：
--
作者：
Al-Khayal K;Alafeefy A;Vaali-Mohammed MA;Mahmood A;Zubaidi A;Al-Obeed O;Khan Z;Abdulla M;Ahmad R
结直肠癌(CRC)是世界上第三种最常见的癌症。治疗晚期结直肠癌需要新的抗癌药物，因为大多数死亡是由于癌症转移造成的。新近开发的化合物3c已被证明具有强大的抗肿瘤作用，但其抗肿瘤作用的机制尚不清楚。用四甲基偶氮唑盐比色法和xCELLigence RTCA DP仪检测NAC对HT-29和SW620细胞增殖的抑制作用。用流式细胞仪进行3C诱导的细胞凋亡研究。用荧光平板阅读器和流式细胞仪检测细胞内ROS的生成，用流式细胞仪检测线粒体膜电位，用比色法测定NADPH和GSH水平。Western blotting检测BCL2家族蛋白表达、细胞色素c释放及PARP活性。用酶联免疫吸附试验检测半胱氨酸天冬氨酸蛋白酶活性。采用实时荧光定量xCELLigence RTCA DP系统对SW620细胞进行细胞迁移实验，对HT-29细胞进行伤口愈合实验。许多抗癌药物通过诱导活性氧(ROS)发挥作用。在这项研究中，我们证明了3c诱导的细胞增殖抑制可以被抗氧化剂N-乙酰半胱氨酸逆转，这表明3c通过增加HT-29细胞中ROS的产生来发挥作用。3c处理的结直肠癌细胞中ROS的直接测量证实了这一点。此外，3c还可降低HT-29细胞的NADPH和谷胱甘肽水平。此外，对细胞凋亡途径的研究表明，细胞色素c的释放增加导致半胱氨酸天冬氨酸氨基转移酶-9的激活，进而激活半胱氨酸天冬氨酸氨基转移酶-3和半胱氨酸天冬氨酸氨基转移酶6。3C还能(I)增加p53和−的表达，(Ii)降低bcl2和bclxL的表达，(Iii)诱导PARP裂解。证实了我们的观察结果，NAC显著抑制了细胞凋亡的诱导、ROS的产生、细胞色素c的释放和PARP的裂解。结果进一步表明，3c通过调节EMT标志物和抑制转化生长因子β诱导的Smad2和Samd3的磷酸化来抑制细胞迁移。因此，我们的发现表明，3c破坏了结直肠癌细胞的氧化还原平衡，并支持这种药物可能对结直肠癌有效的观点。本文的在线版本(doi：10.1186/s12885-0163005-7)包含补充材料，授权用户可以使用。
Colorectal cancer (CRC) is the 3rd most common type of cancer worldwide. New anti-cancer agents are needed for treating late stage colorectal cancer as most of the deaths occur due to cancer metastasis. A recently developed compound, 3c has shown to have potent antitumor effect; however the mechanism underlying the antitumor effect remains unknown. 3c-induced inhibition of proliferation was measured in the absence and presence NAC using MTT in HT-29 and SW620 cells and xCELLigence RTCA DP instrument. 3c-induced apoptotic studies were performed using flow cytometry. 3c-induced redox alterations were measured by ROS production using fluorescence plate reader and flow cytometry and mitochondrial membrane potential by flow cytometry; NADPH and GSH levels were determined by colorimetric assays. Bcl2 family protein expression and cytochrome c release and PARP activation was done by western blotting. Caspase activation was measured by ELISA. Cell migration assay was done using the real time xCELLigence RTCA DP system in SW620 cells and wound healing assay in HT-29. Many anticancer therapeutics exert their effects by inducing reactive oxygen species (ROS). In this study, we demonstrate that 3c-induced inhibition of cell proliferation is reversed by the antioxidant, N-acetylcysteine, suggesting that 3c acts via increased production of ROS in HT-29 cells. This was confirmed by the direct measurement of ROS in 3c-treated colorectal cancer cells. Additionally, treatment with 3c resulted in decreased NADPH and glutathione levels in HT-29 cells. Further, investigation of the apoptotic pathway showed increased release of cytochrome c resulting in the activation of caspase-9, which in turn activated caspase-3 and −6. 3c also (i) increased p53 and Bax expression, (ii) decreased Bcl2 and BclxL expression and (iii) induced PARP cleavage in human colorectal cancer cells. Confirming our observations, NAC significantly inhibited induction of apoptosis, ROS production, cytochrome c release and PARP cleavage. The results further demonstrate that 3c inhibits cell migration by modulating EMT markers and inhibiting TGFβ-induced phosphorylation of Smad2 and Samd3. Our findings thus demonstrate that 3c disrupts redox balance in colorectal cancer cells and support the notion that this agent may be effective for the treatment of colorectal cancer. The online version of this article (doi:10.1186/s12885-016-3005-7) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
7.4
作者：
Circu, Magdalena L.;Aw, Tak Yee
通讯作者：
Aw, Tak Yee
靶向碳酸酐酶IX用于癌症治疗剂的最新发展。
影响因子：
--
作者：
McDonald PC;Winum JY;Supuran CT;Dedhar S
通讯作者：
Dedhar S
影响因子：
3.7
作者：
Hussain AR;Uddin S;Bu R;Khan OS;Ahmed SO;Ahmed M;Al-Kuraya KS
通讯作者：
Al-Kuraya KS
影响因子：
9
作者：
通讯作者：
--
影响因子：
3.7
作者：
Larocque K;Ovadje P;Djurdjevic S;Mehdi M;Green J;Pandey S
通讯作者：
Pandey S