组蛋白去乙酰化酶抑制剂JNJ-26481585抗食管癌活性及其作用机制研究
|
摘要
目的研究组蛋白去乙酰化酶(HDAC)抑制剂JNJ-26481585的抗食管癌活性及其分子机制。方法使用溶剂对照和浓度梯度的JNJ-26481585作用于食管癌细胞株TE-1,采用MTT法和克隆形成实验检测处理后的细胞活力;EdU染色检测细胞增殖情况;流式细胞术检测细胞周期和凋亡;划痕和Transwell侵袭实验检测JNJ-26481585的抗转移活性;Western blot实验检测JNJ-26481585对食管癌生长相关信号通路的影响。结果 JNJ-26481585在较低浓度即可有效抑制TE-1细胞的活力、克隆形成和增殖,诱导细胞G_2/M期阻滞和凋亡,同时可抑制细胞迁移和侵袭的活性。Western blot实验结果显示,JNJ-26481585可明显提升TE-1细胞中p21蛋白表达水平,并有效抑制PI3K/mTOR和MAPK通路中的关键蛋白Akt、ERK的磷酸化。结论 HDAC抑制剂JNJ-26481585可通过上调细胞周期抑制剂p21表达,以及抑制Akt/mTOR、ERK信号通路,发挥抗食管癌作用。
Aim To study the anti-esophageal cancer activity of a pan-HDAC inhibitor JNJ-26481585 and its molecular mechanism. Methods Esophageal cancer cell line TE-1 was treated with vehicle control or serial dilutions of JNJ-26481585, and then MTT and colony formation assay, EdU incorporayion assay, and flow cytometry were performed to detect cell viability, cell proliferation, cell cycle and apoptosis, respectively. The anti-metastasis activity of JNJ-26481585 was assessed according to wound healing assay and Transwell invasion assay. Additionally, Western blot was performed to detect the influence of JNJ-26481585 on the growth-related signaling pathways of esophageal cancer. Results JNJ-26481585 effectively inhibited cellular viability and proliferation of TE-1 cells, as well as induced G_2/M phase arrest and apoptosis even at low treatment concentrations. Meanwhile, it also had the ability to suppress the migration and invasion of TE-1 cells. The results of Western blotting indicated that JNJ-26481585 treatment could significantly up-regulate the expression level of P21 protein in TE-1 cells, and inhibit the phosphorylation of Akt and ERK, the pivotal proteins of PI3 K/mTOR and MAPK pathways, respectively. Conclusions JNJ-26481585 exerts its anti-esophageal cancer effects through multiple molecular mechanisms, including up-regulation of cell cycle inhibitor P21 and blockade of Akt/mTOR and ERK signaling cascades.
Aim To study the anti-esophageal cancer activity of a pan-HDAC inhibitor JNJ-26481585 and its molecular mechanism. Methods Esophageal cancer cell line TE-1 was treated with vehicle control or serial dilutions of JNJ-26481585, and then MTT and colony formation assay, EdU incorporayion assay, and flow cytometry were performed to detect cell viability, cell proliferation, cell cycle and apoptosis, respectively. The anti-metastasis activity of JNJ-26481585 was assessed according to wound healing assay and Transwell invasion assay. Additionally, Western blot was performed to detect the influence of JNJ-26481585 on the growth-related signaling pathways of esophageal cancer. Results JNJ-26481585 effectively inhibited cellular viability and proliferation of TE-1 cells, as well as induced G_2/M phase arrest and apoptosis even at low treatment concentrations. Meanwhile, it also had the ability to suppress the migration and invasion of TE-1 cells. The results of Western blotting indicated that JNJ-26481585 treatment could significantly up-regulate the expression level of P21 protein in TE-1 cells, and inhibit the phosphorylation of Akt and ERK, the pivotal proteins of PI3 K/mTOR and MAPK pathways, respectively. Conclusions JNJ-26481585 exerts its anti-esophageal cancer effects through multiple molecular mechanisms, including up-regulation of cell cycle inhibitor P21 and blockade of Akt/mTOR and ERK signaling cascades.
引文
[1] Torre L A, Bray F, Siegel R L, et al. Global cancer statistics, 2012 [J]. CA Cancer J Clin, 2015, 65(2): 87-108.
[2] Goey A K, Sissung T M, Peer C J, et al. Pharmacogenomics and histone deacetylase inhibitors [J]. Pharmacogenomics, 2016, 17(16): 1807-15.
[3] Qin H T, Li H Q, Liu F. Selective histone deacetylase small molecule inhibitors: recent progress and perspectives [J]. Expert Opin Ther Pat, 2017, 27(5): 621-36.
[4] Muller B M, Jana L, Kasajima A, et al. Differential expression of histone deacetylases HDAC1, 2 and 3 in human breast cancer–overexpression of HDAC2 and HDAC3 is associated with clinicopathological indicators of disease progression [J]. BMC Cancer, 2013, 13: 215.
[5] Nakagawa M, Oda Y, Eguchi T, et al. Expression profile of class I histone deacetylases in human cancer tissues [J]. Oncol Rep, 2007, 18(4): 769-74.
[6] Cao L L, Yue Z, Liu L, et al. The expression of histone deacetylase HDAC1 correlates with the progression and prognosis of gastrointestinal malignancy [J]. Oncotarget, 2017, 8(24): 39241-53.
[7] 周慧, 周伟强. 组蛋白去乙酰化酶抑制剂调控p21WAF1/CIP1启动子乙酰化水平影响乳腺癌MCF-7细胞周期 [J]. 中国药理学通报, 2017, 33(10): 1421-5.[7] Zhou H, Zhou W Q. SAHA affects cell cycle of MCF-7 breast cancer cells by regulating acetylated levels of p21WAF1/CIP1 promoter [J]. Chin Pharmacol Bull, 2017, 33(10): 1421-5.
[8] Conte M, De Palma R, Altucci L. HDAC inhibitors as epigenetic regulators for cancer immunotherapy [J]. Int J Biochem Cell Biol, 2018, 98: 65-74.
[9] VicenTE-Dueňas C, Hauer J, Cobaleda C, et al. Epigenetic priming in cancer initiation [J]. Trends Cancer, 2018, 4(6): 408-17.
[10] Wahaib K, Beggs A E, Campbell H, et al. Panobinostat: a histone deacetylase inhibitorfor the treatment of relapsed or refractory multiple myeloma [J]. Am J Health Syst Pharm, 2016, 73(7): 441-50.
[11] Yoon S, Eom G H. HDAC and HDAC inhibitor: from cancer to cardiovascular diseases [J]. Chonnam Med J, 2016, 52: 1-11.
[12] Arts J, King P, Mari?n A, et al. JNJ-26481585, a novel “second-generation” oral histone deacetylase inhibitor, shows broad-spectrum preclinical antitumoral activity [J]. Clin Cancer Res, 2009, 15(22): 6841-51.
[13] Witt O, Sand K, Pekrun A. Butyra TE-induced erythroid differentiation of human K562 leukemia cells involves inhibition of ERK and activation of p38 MAP kinase pathways [J]. Blood, 2000, 95(7): 2391-6.
[14] Zhang P, Guo Z, Wu Y, et al. Histone deacetylase inhibitors inhibit the proliferation of gallbladder carcinoma cells by suppressing AKT/mTOR signaling [J]. PLoS One, 2015, 10(8): e0136193.
[2] Goey A K, Sissung T M, Peer C J, et al. Pharmacogenomics and histone deacetylase inhibitors [J]. Pharmacogenomics, 2016, 17(16): 1807-15.
[3] Qin H T, Li H Q, Liu F. Selective histone deacetylase small molecule inhibitors: recent progress and perspectives [J]. Expert Opin Ther Pat, 2017, 27(5): 621-36.
[4] Muller B M, Jana L, Kasajima A, et al. Differential expression of histone deacetylases HDAC1, 2 and 3 in human breast cancer–overexpression of HDAC2 and HDAC3 is associated with clinicopathological indicators of disease progression [J]. BMC Cancer, 2013, 13: 215.
[5] Nakagawa M, Oda Y, Eguchi T, et al. Expression profile of class I histone deacetylases in human cancer tissues [J]. Oncol Rep, 2007, 18(4): 769-74.
[6] Cao L L, Yue Z, Liu L, et al. The expression of histone deacetylase HDAC1 correlates with the progression and prognosis of gastrointestinal malignancy [J]. Oncotarget, 2017, 8(24): 39241-53.
[7] 周慧, 周伟强. 组蛋白去乙酰化酶抑制剂调控p21WAF1/CIP1启动子乙酰化水平影响乳腺癌MCF-7细胞周期 [J]. 中国药理学通报, 2017, 33(10): 1421-5.[7] Zhou H, Zhou W Q. SAHA affects cell cycle of MCF-7 breast cancer cells by regulating acetylated levels of p21WAF1/CIP1 promoter [J]. Chin Pharmacol Bull, 2017, 33(10): 1421-5.
[8] Conte M, De Palma R, Altucci L. HDAC inhibitors as epigenetic regulators for cancer immunotherapy [J]. Int J Biochem Cell Biol, 2018, 98: 65-74.
[9] VicenTE-Dueňas C, Hauer J, Cobaleda C, et al. Epigenetic priming in cancer initiation [J]. Trends Cancer, 2018, 4(6): 408-17.
[10] Wahaib K, Beggs A E, Campbell H, et al. Panobinostat: a histone deacetylase inhibitorfor the treatment of relapsed or refractory multiple myeloma [J]. Am J Health Syst Pharm, 2016, 73(7): 441-50.
[11] Yoon S, Eom G H. HDAC and HDAC inhibitor: from cancer to cardiovascular diseases [J]. Chonnam Med J, 2016, 52: 1-11.
[12] Arts J, King P, Mari?n A, et al. JNJ-26481585, a novel “second-generation” oral histone deacetylase inhibitor, shows broad-spectrum preclinical antitumoral activity [J]. Clin Cancer Res, 2009, 15(22): 6841-51.
[13] Witt O, Sand K, Pekrun A. Butyra TE-induced erythroid differentiation of human K562 leukemia cells involves inhibition of ERK and activation of p38 MAP kinase pathways [J]. Blood, 2000, 95(7): 2391-6.
[14] Zhang P, Guo Z, Wu Y, et al. Histone deacetylase inhibitors inhibit the proliferation of gallbladder carcinoma cells by suppressing AKT/mTOR signaling [J]. PLoS One, 2015, 10(8): e0136193.