Al-Obeed Omar, Vaali-Mohammed Mansoor-Ali, Eldehna Wagdy M, Al-Khayal Khayal, Mahmood Amer, Abdel-Aziz Hatem A, Zubaidi Ahmed, Alafeefy Ahmed, Abdulla Maha, Ahmad Rehan
Colorectal Research Chair, Department of Surgery, King Khaled University Hospital, College of Medicine, King Saud University, Riyadh, Saudi Arabia.
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Kafrelsheikh University, Kafrelsheikh, Egypt.
Onco Targets Ther. 2018 Jun 5;11:3313-3322. doi: 10.2147/OTT.S148108. eCollection 2018.
Colorectal cancer (CRC) is a major worldwide health problem owing to its high prevalence and mortality rate. Developments in screening, prevention, biomarker, personalized therapies and chemotherapy have improved detection and treatment. However, despite these advances, many patients with advanced metastatic tumors still succumb to the disease. New anticancer agents are needed for treating advanced stage CRC as most of the deaths occur due to cancer metastasis. A recently developed novel sulfonamide derivative 4-((2-(4-(dimethylamino) phenyl)quinazolin-4-yl)amino)benzenesulfonamide (3D) has shown potent antitumor effect; however, the mechanism underlying the antitumor effect remains unknown.
3D-mediated inhibition on cell viability was evaluated by MTT and real-time cell proliferation was measured by xCelligence RTDP instrument. Western blotting was used to measure pro-apoptotic, anti-apoptotic proteins and JAK2-STAT3 phosphorylation. Flow cytometry was used to measure ROS production and apoptosis.
Our study revealed that 3D treatment significantly reduced the viability of human CRC cells HT-29 and SW620. Furthermore, 3D treatment induced the generation of reactive oxygen species (ROS) in human CRC cells. Confirming our observation, N-acetylcysteine significantly inhibited apoptosis. This is further evidenced by the induction of p53 and Bax; release of cytochrome c; activation of caspase-9, caspase-7 and caspase-3; and cleavage of PARP in 3D-treated cells. This compound was found to have a significant effect on the inhibition of antiapoptotic proteins Bcl2 and BclxL. The results further demonstrate that 3D inhibits JAK2-STAT3 pathway by decreasing the constitutive and IL-6-induced phosphorylation of STAT3. 3D also decreases STAT3 target genes such as cyclin D1 and survivin. Furthermore, a combination study of 3D with doxorubicin (Dox) also showed more potent effects than single treatment of Dox in the inhibition of cell viability.
Taken together, these findings indicate that 3D induces ROS-mediated apoptosis and inhibits JAK2-STAT3 signaling in CRC.
由于结直肠癌（CRC）的高发病率和死亡率，它是一个全球性的主要健康问题。筛查、预防、生物标志物、个性化治疗和化疗方面的进展改善了疾病的检测和治疗。然而，尽管有这些进展，许多晚期转移性肿瘤患者仍然死于该疾病。由于大多数死亡是由癌症转移导致的，因此需要新的抗癌药物来治疗晚期结直肠癌。最近开发的一种新型磺胺衍生物4-((2-(4-(二甲基氨基)苯基)喹唑啉-4-基)氨基)苯磺酰胺（3D）已显示出强大的抗肿瘤作用；然而，其抗肿瘤作用的潜在机制仍然未知。
通过MTT评估3D对细胞活力的抑制作用，并使用xCelligence RTDP仪器测量实时细胞增殖。采用蛋白质免疫印迹法检测促凋亡蛋白、抗凋亡蛋白以及JAK2-STAT3磷酸化水平。使用流式细胞术检测活性氧（ROS）的产生和细胞凋亡情况。
我们的研究表明，3D处理显著降低了人结直肠癌细胞HT-29和SW620的活力。此外，3D处理诱导了人结直肠癌细胞中活性氧（ROS）的生成。N-乙酰半胱氨酸显著抑制凋亡，证实了我们的观察结果。3D处理的细胞中p53和Bax的诱导、细胞色素c的释放、caspase-9、caspase-7和caspase-3的激活以及PARP的裂解进一步证明了这一点。发现该化合物对抑制抗凋亡蛋白Bcl2和BclxL有显著作用。结果进一步表明，3D通过降低STAT3的组成型和IL-6诱导的磷酸化来抑制JAK2-STAT3通路。3D还降低了STAT3靶基因如细胞周期蛋白D1和存活素的表达。此外，3D与阿霉素（Dox）的联合研究在抑制细胞活力方面也显示出比单独使用Dox更强大的效果。
综上所述，这些发现表明3D在结直肠癌中诱导ROS介导的凋亡并抑制JAK2-STAT3信号传导。
