米非司酮、米索前列醇对滋养细胞TRIM22表达的影响

doi:10.6040/j.issn.1671-7554.0.2019.344

摘要/Abstract

摘要： 目的探讨米非司酮、米索前列醇对滋养细胞TRIM22表达的影响。方法临床病例分组如下:将40例早期妊娠患者分为胚胎停育组(A组)12例,人工流产组(B组)14例,药物流产组(C组)14例;体外培养滋养细胞HTR-8,按2×2析因设计随机分为如下4组:空白对照组(D组)、米索前列醇组(E组)、米非司酮组(F组)、米非司酮+米索前列醇组(G组)。采用CCK-8法检测各组细胞的增殖能力。细胞划痕实验检测各组细胞的迁移能力。免疫组化检测TRIM22蛋白表达部位及表达强度。实时荧光定量PCR和Western blottiing检测TRIM22 mRNA 及蛋白表达水平并进行半定量分析。结果临床病例中,TRIM22主要表达在滋养细胞的细胞核;A、C两组的TRIM22蛋白和mRNA表达强度低于B组,差异有统计学意义(P<0.05),而A、C两组间差异无统计学意义(P>0.05)。滋养细胞体外模型中,使用米非司酮组(F、G组)与未使用米非司酮组(D、E组)间比较,滋养细胞增殖能力和迁移能力降低,TRIM22 mRNA和蛋白表达强度降低(P<0.001);使用米索前列醇组(E、G组)与未使用米索前列醇组(D、F组)间比较,差异无统计学意义(P>0.05);米非司酮与米索前列醇间无明显交互作用(P>0.05)。结论米非司酮可能通过下调TRIM22的表达抑制了滋养细胞的增殖、迁移,米索前列醇对其作用无统计学意义影响。

关键词: TRIM22, 米非司酮, 米索前列醇, 滋养细胞, 药物流产

Abstract: Objective To investigate the effects of mifepristone and misoprostol on the expression of TRIM22 in trophoblast cells. Methods Forty cases of early pregnancy were divided into embryo aborting group(group A, n=12), artificial abortion group(group B, n=14)and drug abortion group(group C, n=14). HTR-8 cells were cultured in vitro and randomly divided into blank control group(group D), misoprostol group(group E), mifepristone group(group F), and mifepristone plus misoprostol group(group G)according to 2×2 factorial design. CCK-8 assay was used to detect the proliferation of cells, and cell scratch test was used to detect the migration ability of cells. The expression site and intensity of TRIM22 protein were detected by immunohistochemistry. Real-time fluorescence quantitative PCR and Western blotting were used to detect the expression of TRIM22 mRNA and protein and semi-quantitative analysis was performed. Results In the clinical cases, TRIM22 was mainly expressed in the nucleus of trophoblasts; the expression intensity of TRIM22 protein and mRNA in groups A and C was lower than that in group B(P<0.05), but there was no 山东大学学报 (医学版)57卷10期 -赵立美,等.米非司酮、米索前列醇对滋养细胞TRIM22表达的影响 \=- significant difference between groups A and C (P>0.05). For the in vitro model of trophoblasts, the proliferation and migration ability of trophoblasts, and the expressions of TRIM22 protein and mRNA in the groups using mifepristone(group F, group G)were decreased compared with the groups without mifepristone(group D, group E)(P<0.001). There was no significant difference between misoprostol groups(group E, group G)and non-misoprostol group(group D, group F)(P>0.05); there was no significant interaction between mifepristone and misoprostol (P>0.05). Conclusion Mifepristone may inhibit the proliferation and migration of trophoblasts by down-regulating the expression of TRIM22. Misoprostol has no significant effect on the proliferation and migration of trophoblasts.

Key words: TRIM22, Mifepristone, Misoprostol, Trophoblast cells, Drug abortion

中图分类号:

R714

赵立美,颜磊,申晓畅,孙一卿,何鹏娟,赵兴波. 米非司酮、米索前列醇对滋养细胞TRIM22表达的影响[J]. 山东大学学报 (医学版), 2019, 57(10): 86-92.

ZHAO Limei, YAN Lei, SHEN Xiaochang, SUN Yiqing, HE Pengjuan, ZHAO Xingbo. Effects of mifepristone and misoprostol on the expression of TRIM22 in trophoblast cells[J]. Journal of Shandong University (Health Sciences), 2019, 57(10): 86-92.

参考文献

[1] Versteeg GA, Rajsbaum R, Sánchezaparicio MT, et al. The E3-ligase TRIM family of proteins regulates signaling pathways triggered by innate immune pattern-recognition receptors[J]. Immunity, 2013, 38(2): 384-398.
[2] Rajsbaum R, García-Sastre A, Versteeg GA. TRIMmunity: the roles of the TRIM E3-ubiquitin ligase family in innate antiviral immunity[J]. J Mol Biol, 2014, 426(6): 1265-1284.
[3] Nautiyal J, Kumar PG, Laloraya M. Mifepristone(Ru486)antagonizes monocyte chemotactic protein-3 down-regulation at early mouse pregnancy revealing immunomodulatory events in Ru486 induced abortion[J]. Am J Reprod Immunol, 2015, 52(1): 8-18.
[4] Mao G, Wang J, Kang Y, et al. Progesterone increases systemic and local uterine proportions of CD4+CD25+ Treg cells during midterm pregnancy in mice[J]. Endocrinology, 2010, 151(11): 5477-5488.
[5] Cechin SR, Buchwald P. Effects of representative glucocorticoids on TNFα- and CD40L-induced NF-κB activation in sensor cells[J]. Steroids, 2014, 85(33): 36-43.
[6] Danielsson KG, Marions L, Bygdeman M. Effects of mifepristone on endometrial receptivity[J]. Steroids, 2003, 68(10): 1069-1075.
[7] 孙颖,郑晓静,李明江,等.米非司酮对早孕胎盘绒毛CEACAM6表达的影响[J].山东医药, 2013, 53(1): 43-44.
[8] 于鹏,颜磊,王飞,等.米非司酮对体外培养绒毛组织SPAG9表达的影响[J]. 医学与哲学(临床决策论坛版), 2011, 32(1): 50-51. YU Peng, YAN Lei, WANG Fei, et al. Effect of mifepristone on the expression of SPAG9 in villous explants[J]. Medicine and Philosophy(Clinical Decision Making Forum Edition), 2011, 32(1): 50-51.
[9] 鹿群,赵兴波,李建峰.米非司酮对人早孕绒毛、蜕膜组织血管生成的影响探讨[J].中国计划生育学杂志, 2008(2): 82-85. doi: 10.3969/j.issn.1004-8189.2008.02.008. LU Qun, ZHAO Xingbo, LI Jianfeng. Probe into the effect of mifepristone on angiogenesis of human chorionic villi and decidual cells[J]. Chinese Journal of Family Planning, 2008(2): 82-85. doi: 10.3969/j.issn.1004-8189.2008.02.008.
[10] 韩雪川,李明江,赵兴波,等.米非司酮对人早孕绒毛组织中TGFβ/smads信号通路功能状态的影响[J].山东大学学报(医学版), 2009, 47(7): 93-95. HAN Xuechuan, LI Mingjiang, ZHAO Xingbo, et al. Mifepristone modulates the function of TGF-β smads signaltransduction pathway in early pregnant chorionic villi[J]. Journal of Shandong University(Health Sciences), 2009, 47(7): 93-95.
[11] 郑晓静,孙颖,刘姝,等.米非司酮对人早孕绒毛组织核转录因子-κB表达的影响[J].山东大学学报(医学版), 2010, 48(1): 106-108. ZHENG Xiaojing, SUN Ying, LIU Shu, et al. Effect of mifepristone on expression of nuclear factor-κB in human villi during early pregnancy[J]. Journal of Shandong University(Health Sciences), 2010, 48(1): 106-108.
[12] Tocchini C, Ciosk R. TRIM-NHL proteins in development and disease[J]. Semin Cell Dev Biol, 2015(47-48): 52-59. doi: 10.1016/j.semcdb.2015.10.017.
[13] Uchil PD, Hinz A, Siegel S, et al. TRIM protein-mediated regulation of inflammatory and innate immune signaling and its association with antiretroviral activity[J]. J Virol, 2013, 87(1): 257-272.
[14] Wu X, Anderson JL, Campbell EM, et al. Proteasome inhibitors uncouple rhesus TRIM5alpha restriction of HIV-1 reverse transcription and infection[J]. Proc Natl Acad Sci U S A, 2006, 103(19): 7465-7470.
[15] Tocchini C, Keusch JJ, Miller SB, et al. The TRIM-NHL protein LIN-41 controls the onset of developmental plasticity in caenorhabditis elegans[J]. PLoS Genet, 2014, 10(8): e1004533. doi: 10.1371/journal.pgen.1004533.
[16] Li H, Sun L, Tang Z, et al. Overexpression of TRIM24 correlates with tumor progression in non-small cell lung cancer[J]. PLoS One, 2012, 7(5): e37657. doi: 10.1371/journal.pone.0037657.
[17] Zaman MU, Nomura T, Takagi T, et al. Ubiquitination-deubiquitination by the TRIM27-USP7 complex regulates tumor necrosis factor alpha-induced apoptosis[J]. Mol Cell Biol, 2013, 33(24): 4971-4984.
[18] Horie-Inoue K. TRIM proteins as trim tabs for the homoeostasis[J]. J Biochem, 2013, 154(4): 309-312.
[19] 王雨涛,周济,杨冬,等.TRIM29基因在胶质瘤中的表达及其对胶质母细胞瘤细胞U-87 MG增殖与迁移的影响[J]. 第三军医大学学报, 2018, 40(11): 959-965. WANG Yutao, ZHOU Ji,YANG Dong, et al. Expression of TRIM29 in gliomas and its effect on proliferation and migration of glioblastoma U-87 MG cells[J]. Journal of Third Military Medical University 2018, 40(11): 959-965.
[20] 龙璐,王堃,陈贞,等.三结构域包含蛋白29对鼻咽癌细胞生长和迁移的影响[J].中国现代医学杂志, 2015, 25(2): 7-12. LONG Lu, WANG Kun, CHEN Zhen, et al. Effects of TRIM 29 on growth and migration of nasopharyngeal carcinoma cells[J]. China Journal of Modern Medicine. 2015, 25(2): 7-12.
[21] Zhou XM, Sun R, Luo DH, et al. Upregulated TRIM29 promotes proliferation and metastasis of nasopharyngeal carcinoma via PTEN/AKT/mTOR signal pathway[J]. Oncotarget, 2016, 7(12): 13634-13650.
[22] Murray MJ, Lessey BA. Embryo implantation and tumor metastasis: common pathways of invasion and angiogenesis[J]. Semin Reprod Endocrinol, 1999, 17(3): 275-290.
[23] 高波,段志坚,徐薇,等.抗病毒固有免疫分子TRIM22 C端SPRY结构域对其转录、翻译及亚细胞定位的影响[J].微生物与感染, 2010, 5(1): 31-35. GAO Bo, DUAN Zhijian, XU Wei, et al. The contribution of the C terminal SPRY domain of TRIM22 to its own transcription, translation, and subcellular localization[J]. Journal of Microbes and Infection, 2010, 5(1): 31-35.
[24] Saito-Kanatani M, Urano T, Hiroi H, et al. Identification of TRIM22 as a progesterone-responsive gene in Ishikawa endometrial cancer cells[J]. J Steroid Biochem Mol Biol, 2015, 154: 217-225. doi: 10.1016/j.jsbmb.2015.08.024.
[25] Obad S, Brunnström H, Vallonchristersson J, et al. Staf50 is a novel p53 target gene conferring reduced clonogenic growth of leukemic U-937 cells[J]. Oncogene, 2004, 23(23): 4050.
[26] Sun Y, Ho GH, Koong HN, et al. Down-regulation of Tripartite-motif containing 22 expression in breast cancer is associated with a lack of p53-mediated induction[J]. Biochem Biophys Res Commun, 2013, 441(3): 600-606.
[27] Zirn B, Hartmann O, Samans B, et al. Expression profiling of Wilms tumors reveals new candidate genes for different clinical parameters[J]. Int J Cancer, 2010, 118(8): 1954-1962.
[28] Vicent GP, Zaurin R, Nacht AS, et al. Nuclear factor 1 synergizes with progesterone receptor on the mouse mammary tumor virus promoter wrapped around a histone H3/H4 tetramer by facilitating access to the central hormone-responsive elements[J]. J Biol Chem, 2010, 285(4): 2622-2631.

多维度评价

Viewed

Full text

Abstract

Cited

Shared

Discussed