内质网相关降解蛋白Derlin-1通过抑制ERK信号通路抗人结肠癌细胞增殖

doi:10.12092/j.issn.1009-2501.2018.12.006

摘要/Abstract

摘要：

目的: 探讨内质网相关降解蛋白Derlin-1对人结肠癌SW480细胞凋亡的影响及其可能的作用机制。方法: 采用sh-Derlin-1-pGPU6/Neo质粒转染构建瞬时敲除Derlin-1基因的SW480细胞株，应用实时定量PCR和免疫印迹验证Derlin-1基因和蛋白的低表达，采用CCK8法检测SW480细胞增殖情况，流式细胞仪检测SW480细胞凋亡情况，Western blot检测SW480细胞中凋亡蛋白Bcl-2、Bax表达量和ERK磷酸化水平。结果: 使用pGPU6/Neo载体能够稳定地抑制SW480细胞中Derlin-1的表达，sh-Derlin-1组细胞增殖率相对于对照组显著降低、凋亡率显著升高（P<0.05），Western blot实验结果发现sh-Derlin-1组细胞中Bcl-2表达量显著降低、Bax表达量显著升高及ERK磷酸化水平显著降低，与对照组相比差异有统计学意义（P<0.05）。结论: Derlin-1表达的降低能抑制ERK信号通路活化，进而诱导人结肠癌SW480细胞凋亡、抑制增殖。

Abstract:

AIM: To observe the effects of endoplasmic reticulum associated degradation protein Derlin-1 on the apoptosis of human colon cancer cells (SW480) and its possible mechanism. METHODS: The qualified recombinant plasmid sh-Derlin-1-pGPU6/Neo was transfected into SW480 cells with Derlin-1 knocking down. The expression level of Derlin-1 was detected at both mRNA and protein levels by real-time PCR and Western blot. CCK8 assay and flow cytometry were performed to detect the proliferation and apoptosis of SW480 cells. We examined the expression of apoptosis associated proteins (Bcl-2 and Bax) and ERK phosphorylation followed by Derlin-1 low-expression by Western blot. RESULTS: The Derlin-1 knocking down plasmid was constructed successfully. SW480 cells with Derlin-1 knocking down had been established successfully. Compared with control group, the proliferation rates of sh-Derlin-1 SW480 cells were significantly decreased, while the apoptotic rates of sh-Derlin-1 SW480 cells were significantly increased (P<0.05); the expression levels of Bcl-2 and p-ERK in sh-Derlin-1 group were significantly decreased, while the expression level of Bax was greatly increased (P<0.05). CONCLUSION: The decreased level of Derlin-1 can suppress the proliferation and induce the apoptosis of SW480 cells through suppressing the activity of ERK signaling pathway.

Key words: Derlin-1, human colon cancer cells, proliferation, apoptosis, ERK signaling pathway

中图分类号:

R965.2

赵丽，华夏，谭晔. 内质网相关降解蛋白Derlin-1通过抑制ERK信号通路抗人结肠癌细胞增殖[J]. 中国临床药理学与治疗学, 2018, 23(12): 1353-1358.

ZHAO Li, HUA Xia, TAN Ye. Effects of endoplasmic reticulum associated degradation protein Derlin-1 on the proliferation of human colon cancer cells via suppressing ERK pathway[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(12): 1353-1358.

参考文献

［1］赵建国，叶万立，吴东平, 等. 替吉奥联合奥沙利铂治疗晚期结肠癌前后血浆miR-21 表达水平变化及其临床意义［J］. 中国临床药理学与治疗学, 2016, 21(3) : 318-321.
［2］Patel SS, Nelson R, Sanchez J, et al. Elderly patients with colon cancer have unique tumor characteristics and poor survival ［J］. Cancer, 2013, 119(4): 739-747.
［3］张轶雯,楼倩雯,钟里科,等.基于GEO数据库的近端与远端结肠癌特征基因生物信息学研究［J］.中国临床药理学与治疗学,2018,23(4):422-427.
［4］Chen SF, Wu CH, Lee YM, et al. Caveolin-1 interacts with Derlin-1 and promotes ubiquitination and degradation of cyclooxygenase-2 via collaboration with p97 complex［J］. J Biol Chem, 2013, 288(12): 33462-33469.
［5］Wang F, Olson EM, Shyng SL. Role of Derlin-1 protein in proteostasis regulation of ATP-sensitive potassium channels ［J］. J Biol Chem, 2012, 287(5): 10482-10493.
［6］Suzuki M, Otsuka T, Ohsaki Y, et al. Derlin-1 and UBXD8 are engaged in dislocation and degradation of lipidated ApoB-100 at lipid droplets［J］. Mol Biol Cell, 2012, 23(11): 800-810.
［7］Dong Q, Fu L, Zhao Y, et al. Derlin-1 overexpression confers poor prognosis in muscle invasive bladder cancer and contributes to chemoresistance and invasion through PI3K/AKT and ERK/MMP signaling ［J］. Oncotarget, 2017, 8(10): 17059-17069.
［8］Ran Y, Jiang Y, Zhong X, et al. Identification of derlin-1 as a novel growth factor-responsive endothelial antigen by suppression subtractive hybridization ［J］. Biochem Biophys Res Commun, 2016, 348(4): 1272-1278.
［9］Dong QZ, Wang Y, Tang ZP, et al. Derlin-1 is overexpressed in non-small cell lung cancer and promotes cancer cell invasion via EGFR-ERK-mediated up-regulation of MMP-2 and MMP-9［J］. Am J Pathol, 2013, 182: 954-964.
［10］Wang J, Hua H, Ran Y, et al. Derlin-1 is overexpressed in human breast carcinoma and protects cancer cells from endoplasmic endoplasmic reticulum stress-induced apoptosis ［J］. Breast Cancer Res, 2016, 10(7): 7-16.
［11］谭学明, 张丹丹, 唐辉, 等. Derlin-1蛋白在结肠癌组织中的表达及其意义［J］. 现代医学, 2016, 44(8): 1060-1063.
［12］Tan X, He X, Jiang Z, et al. Derlin-1 is overexpressed in human colon cancer and promotes cancer cell proliferation ［J］. Mol Cell Biochem, 2015, 408(1/2): 205-213.
［13］Zhao L, Ackerman SL. Endoplasmic reticulum stress in health and disease ［J］. Curr Opin Cell Biol, 2006, 18(4): 444-452.
［14］Salto R, Vílchez JD, Cabrera E, et al. Activation of ERK by sodium tungstate induces protein synthesis and prevents protein degradation in rat L6 myotubes ［J］. FEBS Lett, 2014, 588(14): 2246-2254.
［15］Xu L, Wang ZH, Xu D, et al. Expression of Derlin-1 and its effect on expression of autophagy marker genes under endoplasmic reticulum stress in lung cancer cells ［J］. Cancer Cell Int, 2014, 14(5): 50-59.
［16］Pi L, Zhu G, She L, et al. Elevated expression of Derlin-1 associates with unfavorable survival time of squamous cell carcinoma of the head and neck and promotes its malignance［J］. J Cancer, 2017, 8(12): 2336-2345.
［17］Dong Q, Fu L, Zhao Y, et al. Derlin-1 overexpression confers poor prognosis in muscle invasive bladder cancer and contributes to chemoresistance and invasion through PI3K/AKT and ERK/MMP signaling ［J］. Oncotarget, 2017, 8(10): 17059-17069.
［18］Yang F, Wei K, Qin Z, et al. MiR-598 Suppresses invasion and migration by negative regulation of Derlin-1 and epithelial-mesenchymal transition in non-small cell lung cancer［J］. Cell Physiol Biochem, 2018, 47(1): 245-256.
［19］Mao M, Zhang J, Jiang J. Overexpression of Derlin-1 is associated with poor prognosis in patients with non-small cell lung cancer ［J］. Ann Clin Lab Sci, 2018, 48(1): 29-34.
［20］Wu Z, Wang C, Zhang Z, et al. High expression of Derlin-1 is associated with the malignancy of bladder cancer in a Chinese Han Population ［J］. PLoS One, 2016, 11(12): e0168351.

[1]	范杰, 金永明, 江孝龙, 蒋国华. lncRNA HOTAIR调控miR-206影响类风湿关节炎滑膜细胞增殖和凋亡的分子机制研究[J]. 中国临床药理学与治疗学, 2023, 28(7): 736-742.
[2]	王安婧, 王亚亚, 梁轩, 闫雅婕, 苏静, 李彩东. 基于PPAR治疗胆汁淤积性肝病的机制与药物研究进展[J]. 中国临床药理学与治疗学, 2023, 28(7): 796-808.
[3]	杨晓佳, 江萌, 吴敏, 张伊黎, 吕兰, 吴嫄芬, 王鑫昱, 刘立权. 藏红花素介导DKK3调控GSK-3β/β-catenin通路对阿尔兹海默症大鼠的认知改善作用[J]. 中国临床药理学与治疗学, 2023, 28(5): 489-497.
[4]	王艳, 张金辉, 赵永辰, 刘宏祥, 刘亚维, 苗欢欢, 杨信才. 红杉黄酮通过下调PI3K/AKT信号通路抑制胃癌细胞增殖侵袭等干细胞特性[J]. 中国临床药理学与治疗学, 2023, 28(5): 508-513.
[5]	侯小煜, 冷玉芳, 曹雪芬, 吕兴娇, 韩晓霞, Janvier NIBARUTA, 刘永强. 五味子乙素减轻小鼠肠缺血再灌注损伤的网络药理学分析及实验验证[J]. 中国临床药理学与治疗学, 2023, 28(2): 147-154.
[6]	王剑, 孙永东, 周兴玮, 刘雷, 陈龙, 童兴科, 朱佳丽. 黄芩素经由miR-125b-5p/IRF4轴进而促进喉癌细胞死亡并抑制其侵袭的机制研究[J]. 中国临床药理学与治疗学, 2023, 28(11): 1209-1218.
[7]	朱瑞芳, 郭东凯, 智慧, 江翊国, 张悦翎, 钱晓萍, 季士亮. ROS-NF-κB-p38MAPK通路探索榄香烯联合硼替佐米抗多发性骨髓瘤的机制研究[J]. 中国临床药理学与治疗学, 2023, 28(11): 1219-1226.
[8]	李菲, 丁慧琴, 陈梦静. 异莲心碱激活ERK信号通路诱导肺癌PC9细胞自噬的作用研究[J]. 中国临床药理学与治疗学, 2023, 28(11): 1235-1240.
[9]	周欢, 徐铭, 李波, 刘春英, 王淳. PI3K/Akt/FOXO3a信号通路介导的保护性自噬参与肺腺癌获得性顺铂耐药表型重塑[J]. 中国临床药理学与治疗学, 2022, 27(9): 961-970.
[10]	华海燕, 严杰, 秦宇芬. 五味子乙素通过抑制心肌细胞凋亡减轻大鼠心肌缺血再灌注损伤的实验研究[J]. 中国临床药理学与治疗学, 2022, 27(8): 848-856.
[11]	丁嘉敏, 王友娣, 赵夏丽, 姜根风, 陈思文, 洪程程, 李书勤, 胡卫华. 生长激素和维生素E联合应用治疗薄型子宫内膜的研究[J]. 中国临床药理学与治疗学, 2022, 27(8): 857-862.
[12]	李雪恒, 李龙. 褪黑素与特发性肺纤维化关系的研究进展[J]. 中国临床药理学与治疗学, 2022, 27(6): 715-720.
[13]	王新丽, 许霞青, 方寒冰, 郭玉忠. 佛波酯增强顺铂的抗肿瘤活性及降低其肾毒性的作用研究[J]. 中国临床药理学与治疗学, 2022, 27(5): 535-543.
[14]	杨柳, 罗爱林, 李世勇. 全麻药物损伤发育期大脑的研究进展[J]. 中国临床药理学与治疗学, 2022, 27(12): 1381-1390.
[15]	谭峰浪, 任宏丽, 王宏飞. 黄柏提取物对Aβ_1-42诱导的海马神经细胞的影响[J]. 中国临床药理学与治疗学, 2022, 27(11): 1213-1220.