Haque Abedul, Rahman Mohammad A, Fuchs James R, Chen Zhuo Georgia, Khuri Fadlo R, Shin Dong M, Amin A R M Ruhul
Department of Hematology and Medical Oncology, Winship Cancer Institute of Emory University, Atlanta, GA 30322, USA.
Deaprtment of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, Ohio State University, Columbus, OH, USA.
Cancer Lett. 2015 Jul 28;363(2):166-75. doi: 10.1016/j.canlet.2015.04.017. Epub 2015 Apr 24.
Unlike chemotherapy drugs, the safety of natural compounds such as curcumin has been well established. However, the potential use of curcumin in cancer has been compromised by its low bioavailability, limited tissue distribution and rapid biotransformation leading to low in vivo efficacy. To circumvent these problems, more potent and bioavailable analogs have been synthesized. In the current study, we investigated the mechanism of anti-tumor effect of one such analog, FLLL12, in lung cancers. IC50 values measured by sulforhodamine B (SRB) assay at 72 h and apoptosis assays (annexin V staining, cleavage of PARP and caspase-3) suggest that FLLL12 is 5-10-fold more potent than curcumin against a panel of premalignant and malignant lung cancer cell lines, depending on the cell line. Moreover, FLLL12 induced the expression of death receptor-5 (DR5). Ablation of the expression of the components of the extrinsic apoptotic pathway (DR5, caspase-8 and Bid) by siRNA significantly protected cells from FLLL12-induced apoptosis (p < 0.05). Analysis of mRNA expression revealed that FLLL-12 had no significant effect on the expression of DR5 mRNA expression. Interestingly, inhibition of global phosphatase activity as well as protein tyrosine phosphatases (PTPs), but not of alkaline phosphatases, strongly inhibited DR5 expression and significantly inhibited apoptosis (p < 0.05), suggesting the involvement of PTPs in the regulation of DR5 expression and apoptosis. We further showed that the apoptosis is independent of p53 and p73. Taken together, our results strongly suggest that FLLL12 induces apoptosis of lung cancer cell lines by posttranscriptional regulation of DR5 through activation of protein tyrosine phosphatase(s).
与化疗药物不同,姜黄素等天然化合物的安全性已得到充分证实。然而,姜黄素在癌症治疗中的潜在应用受到其低生物利用度、有限的组织分布和快速生物转化的影响,导致体内疗效较低。为了克服这些问题,人们合成了更有效且生物利用度更高的类似物。在本研究中,我们研究了一种这样的类似物FLLL12在肺癌中的抗肿瘤作用机制。通过磺酰罗丹明B(SRB)测定法在72小时测量的IC50值以及凋亡测定法(膜联蛋白V染色、PARP和caspase-3的切割)表明,根据细胞系的不同,FLLL12对一组癌前和恶性肺癌细胞系的效力比姜黄素高5至10倍。此外,FLLL12诱导死亡受体-5(DR5)的表达。通过小干扰RNA(siRNA)消除外源性凋亡途径(DR5、caspase-8和Bid)成分的表达可显著保护细胞免受FLLL12诱导的凋亡(p < 0.05)。mRNA表达分析显示,FLLL-12对DR5 mRNA表达没有显著影响。有趣的是,抑制全局磷酸酶活性以及蛋白质酪氨酸磷酸酶(PTP),而不是碱性磷酸酶,强烈抑制DR5表达并显著抑制凋亡(p < 0.05),这表明PTP参与DR5表达和凋亡的调节。我们进一步表明,凋亡与p53和p73无关。综上所述,我们的结果强烈表明,FLLL12通过激活蛋白质酪氨酸磷酸酶对DR5进行转录后调节,从而诱导肺癌细胞系凋亡。