迷迭香酸类似物-11通过抑制ERK/MAPK通路诱导人胃癌MGC-803细胞凋亡
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摘要
目的对迷迭香酸类似物-11(rosmarinic acid analogue-11,RAA-11)诱导人胃癌MGC-803细胞凋亡及其机制进行初步探究。方法 MTT法和克隆形成法观察RAA-11对人胃癌MGC-803细胞增殖的抑制作用;Hoechst 33258染色法观察RAA-11对人胃癌MGC-803细胞核凋亡形态学的影响;流式细胞术检测RAA-11对人胃癌MGC-803细胞凋亡率的影响;Western blot观察RAA-11对人胃癌MGC-803细胞凋亡相关蛋白caspase-3、Bcl-2、Bax及通路蛋白ERK、p-ERK表达的影响。结果 MTT结果提示RAA-11可以抑制人胃癌MGC-803细胞增殖(P<0.01),且呈时间浓度依赖性;克隆形成实验结果提示,RAA-11能抑制人胃癌MGC-803细胞的集落形成能力;Hoechst 33258染色结果提示,RAA-11干预人胃癌MGC-803细胞48 h后,细胞核呈现典型凋亡形态学改变;流式细胞术结果提示,RAA-11对人胃癌MGC-803细胞有明显的促凋亡作用; Western blot结果显示,RAA-11上调人胃癌MGC-803细胞促凋亡相关蛋白caspase-3、Bax(P<0.01),下调抑凋亡相关蛋白Bcl-2的表达水平,并降低了通路蛋白ERK、p-ERK的表达水平(P<0.01)。结论 RAA-11能抑制人胃癌MGC-803细胞的增殖,并能够诱导凋亡,其机制可能与抑制ERK/MAPK通路有关。
Aim To explore the apoptosis and mechanisms of human gastric cancer cells MGC-803 induced by RAA-11.Methods MTT and colony assays were used to detect the survival of MGC-803 cells treated with RAA-11. The effect of RAA-11 on the apoptotic morphology of MGC-803 cells was observed by Hoechst 33258 staining. The effect of RAA-11 on the apoptosis rate of MGC-803 cells was detected with flow cytometry. The effects of RAA-11 on apoptosis related protein expression of caspase-3, Bcl-2 and Bax as well as pathway proteins ERK and p-ERK in MGC-803 cells were observed by Western blot.Results MTT assay showed that the proliferation of MGC-803 cells was effectively inhibited in a time-and concentration-dependent manner(P<0.01). The result of colony suggested that RAA-11 could inhibit the proliferation of MGC-803 cells. Hoechst 33258 staining indicated that the nucleus of MGC-803 cells had a typical apoptotic morphological change after intervention with RAA-11. The result of flow cytometry suggested that RAA-11 had a significant apoptotic effect on MGC-803 cells. The expressions of Bax and caspase-3 were significantly up-regulated(P<0.01), and the expressions of Bcl-2, ERK and p-ERK were down-regulated(P<0.01) in MGC-803 cells treated with RAA-11.Conclusions RAA-11 inhibits the proliferation and induces apoptosis in MGC-803 cells, which may be related to the inhibition of ERK/MAPK pathway.
Aim To explore the apoptosis and mechanisms of human gastric cancer cells MGC-803 induced by RAA-11.Methods MTT and colony assays were used to detect the survival of MGC-803 cells treated with RAA-11. The effect of RAA-11 on the apoptotic morphology of MGC-803 cells was observed by Hoechst 33258 staining. The effect of RAA-11 on the apoptosis rate of MGC-803 cells was detected with flow cytometry. The effects of RAA-11 on apoptosis related protein expression of caspase-3, Bcl-2 and Bax as well as pathway proteins ERK and p-ERK in MGC-803 cells were observed by Western blot.Results MTT assay showed that the proliferation of MGC-803 cells was effectively inhibited in a time-and concentration-dependent manner(P<0.01). The result of colony suggested that RAA-11 could inhibit the proliferation of MGC-803 cells. Hoechst 33258 staining indicated that the nucleus of MGC-803 cells had a typical apoptotic morphological change after intervention with RAA-11. The result of flow cytometry suggested that RAA-11 had a significant apoptotic effect on MGC-803 cells. The expressions of Bax and caspase-3 were significantly up-regulated(P<0.01), and the expressions of Bcl-2, ERK and p-ERK were down-regulated(P<0.01) in MGC-803 cells treated with RAA-11.Conclusions RAA-11 inhibits the proliferation and induces apoptosis in MGC-803 cells, which may be related to the inhibition of ERK/MAPK pathway.
引文
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[14] 韦立群, 徐成飞, 李婉婷, 等. NF-κB和MAPK信号通路参与金雀异黄酮诱导乳腺癌MDA-MB-231细胞凋亡的体外研究[J]. 中国药理学通报, 2018, 34(5): 690-4. Wei L Q, Xu C F, Li W T, et al. NF-κB and MAPK signaling pathway were involved of MDA-MB-231 cell apoptosis in breast cancer induced by genistein in vitro studies[J]. Chin Pharmacol Bull, 2018, 34(5): 690-4.
[15] 邹佩良, 张秋红, 周建甫, 等. 重楼皂苷Ⅰ通过ERK/P65/DNMT1通路抑制前列腺癌PC3细胞生长的分子机制[J]. 中华男科学杂志, 2018, 24(3): 199-205. Zou P L, Zhang Q H, Zhou J F, et al. Molecular mechanism to inhibit the growth of PC3 cells in prostate cancer with the polyphyllinⅠby ERK/P65/DNMT1 pathways[J]. J Chin Male Sci, 2018, 24(3): 199-205.
[2] Wang X T, Xie Y B, Xiao Q. siRNA targeting of Cdx2 inhibits growth of human gastric cancer MGC-803 cells[J]. World J Gastroenterol, 2012, 18(16):1903-14.
[3] Zhu Y W, Yan J K, Li J J, et al. Knockdown of radixin suppresses gastric cancer metastasis in vitro by up-regulation of E-cadherin via NF-κB/Snail pathway[J]. Cell Physiol Biochem, 2016, 39(6):2509-21.
[4] Li Y, Ye J, Chen Z, et al. Annonaceous acetogenins mediated up-regulation of Notch2 exerts growth inhibition in human gastric cancer cells in vitro[J]. Oncotarget, 2017, 8(13):21140-52.
[5] Sotnikova R, Okruhlicova L, Vlkovicova J, et al. Rosmarinic acid administration attenuates diabetes-induced vascular dysfunction of the rat aorta[J]. J Pharm Pharmacol, 2013, 65(5):713-23.
[6] Zhu F, Asada T, Sato A, et al. Rosmarinic acid extract for antioxidant, antiallergic, and alpha-glucosidase inhibitory activities, isolated by supramolecular technique and solvent extraction from Perilla leaves[J]. J Agric Food Chem, 2014, 62(4):885-92.
[7] 韦立群, 李清, 甘嘉亮, 等. 迷迭香酸衍生物RAD-9通过PI3K/Akt和p38 MAPK信号通路诱导胃癌MGC-803细胞凋亡[J]. 中国药理学通报, 2018,34(2): 256-60. Wei L Q, Li Q, Gan J L, et al. Rosemary acid derivatives RAD-9 through PI3K/Akt and p38 MAPK signaling pathway induced MGC-803 gastric cancer cell apoptosis[J]. Chin Pharmacol Bull, 2018,34(2): 256-60.
[8] 李玉龙, 张雪, 廖情谊, 等. 迷迭香酸的合成途径及药理作用研究进展[J]. 现代农业科技, 2016(23):66-7. Li Y L, Zhang X, Liao Q Y, et al. Research progress on the synthesis pathway and pharmacological action of rosemary acid[J]. Mod Agric Technol, 2016(23): 66-7.
[9] 齐锐, 董岩. 迷迭香的化学成分与药理作用研究进展[J]. 广州化工, 2012, 40(11): 43-4, 66. Qi R, Dong Y. Research on the chemical composition and pharmacological action of rosemary[J]. Guangzhou Chem, 2012, 40(11): 43-4, 66.
[10] 冯茹, 黄雯雯, 蒙宁, 等. 迷迭香酸抗恶性肿瘤作用研究进展[J]. 四川生理科学杂志, 2015, 37(3):132-5. Feng R, Huang W W, Meng N, et al. Research progress on the anti-malignant tumor of rosemary[J]. Sichuan J Physiol Sci, 2015, 37(3): 132-5.
[11] Giansanti V, Torriglia A, Scovassi A I. Conversation between apoptosis and autophagy: “Is it your turn or mine?”[J]. Apoptosis, 2011, 16(4): 321-33.
[12] 王宇锋, 陈超, 杨春, 等. 6-姜烯酚诱导人结直肠癌细胞凋亡及与Bax、Bcl-2、Caspase3和PARP1基因表达的影响[J]. 现代食品科技, 2017, 33(11): 7-15. Wang Y F, Chen C, Yang C, et al. 6-gingerol induced apoptosis of colorectal cancer cells and its effects with Bax, Bcl-2, Caspase3 and PARP1 gene expression[J]. Mod Food Technol, 2017, 33(11):7-15.
[13] 陈豫. 骆驼蓬子提取物β-咔啉类生物碱对人胃癌SGC-7901细胞凋亡及PTEN, ERK mRNA表达的影响[D]. 乌鲁木齐:新疆医科大学, 2015. Chen Y. Effect of β-carboline alkaloid extract of Peganum harmala L on apoptosis of human gastric cancer SGC-7901 cells and expression of PTEN and ERK mRNA[D]. Urumqi: Xinjiang Medical University, 2015.
[14] 韦立群, 徐成飞, 李婉婷, 等. NF-κB和MAPK信号通路参与金雀异黄酮诱导乳腺癌MDA-MB-231细胞凋亡的体外研究[J]. 中国药理学通报, 2018, 34(5): 690-4. Wei L Q, Xu C F, Li W T, et al. NF-κB and MAPK signaling pathway were involved of MDA-MB-231 cell apoptosis in breast cancer induced by genistein in vitro studies[J]. Chin Pharmacol Bull, 2018, 34(5): 690-4.
[15] 邹佩良, 张秋红, 周建甫, 等. 重楼皂苷Ⅰ通过ERK/P65/DNMT1通路抑制前列腺癌PC3细胞生长的分子机制[J]. 中华男科学杂志, 2018, 24(3): 199-205. Zou P L, Zhang Q H, Zhou J F, et al. Molecular mechanism to inhibit the growth of PC3 cells in prostate cancer with the polyphyllinⅠby ERK/P65/DNMT1 pathways[J]. J Chin Male Sci, 2018, 24(3): 199-205.
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