[1]谢丽华.17β-雌二醇对人成骨肉瘤细胞CLCF1基因表达的影响[J].医学信息,2021,34(03):60-62,67.[doi:10.3969/j.issn.1006-1959.2021.03.018]
　XIE Li-hua.Effect of 17β-estradiol on CLCF1 Gene Expression in Human Osteosarcoma Cells[J].Medical Information,2021,34(03):60-62,67.[doi:10.3969/j.issn.1006-1959.2021.03.018]
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17β-雌二醇对人成骨肉瘤细胞CLCF1基因表达的影响()

医学信息[ISSN:1006-1959/CN:61-1278/R]

卷:
34卷
期数:
2021年03期
页码:
60-62,67
栏目:
论著
出版日期:
2021-02-01

文章信息/Info

Title:
Effect of 17β-estradiol on CLCF1 Gene Expression in Human Osteosarcoma Cells
文章编号:
1006-1959（2021）03-0060-04
作者:
谢丽华
（福建省中医药科学院基础研究室，福建 福州 350003）
Author(s):
XIE Li-hua
(Basic Research Laboratory,Fujian Academy of Chinese Medical Sciences,Fuzhou 350003,Fujian,China)
关键词:
17β-雌二醇; 人重组心肌营养素样细胞因子1; 雌激素受体抑制剂; 荧光素酶
Keywords:
17β-estradiol; Human recombinant cardiotrophin-like cytokine 1; Estrogen receptor inhibitor; Luciferase
分类号:
R58
DOI:
10.3969/j.issn.1006-1959.2021.03.018
文献标志码:
A
摘要:
目的 探讨17β-雌二醇对人成骨肉瘤细胞CLCF1基因表达的影响。方法 将骨肉瘤细胞MG-63分为对照组、β-雌二醇低、中、高剂量组，对照组加入10-3mol/L DMSO，将17β-雌二醇用DMSO稀释后，分别加入β-雌二醇低、中、高剂量组，终浓度分别为10-10、10-9、10-8 mol/L，干预MG-63细胞24、48、72 h后，实时荧光定量PCR检测各组MG-63细胞中CLCF1 mRNA的表达情况；构建CLCF1基因启动子荧光素酶报告基因载体，采用10-8 mol/L 17β-雌二醇及雌激素受体抑制剂氟维司群（ICI）干预MG63细胞24 h后，双荧光素酶报告基因系统检测双荧光素酶活性。结果 ①干预24 h，β-雌二醇低、中、高剂量组CLCF1 mRNA的表达均高于对照组，差异有统计学意义（P＜0.05）；干预48 h，仅β-雌二醇高剂量组CLCF1 mRNA的表达高于对照组，差异有统计学意义（P＜0.05）；干预72 h，β-雌二醇低组CLCF1 mRNA的表达低于对照组，差异有统计学意义（P＜0.05），β-雌二醇中、高剂量组CLCF1 mRNA的表达与对照组比较，差异无统计学意义（P＞0.05）；②与对照组相比，17β-雌二醇高剂量组荧光素酶活性提高，差异有统计学意义（P＜0.05）；加入雌激素受体抑制剂ICI后，荧光素酶活性较对照组降低，差异有统计学意义（P＜0.05）。结论 17β-雌二醇可通过提高CLCF1启动子活性促进MG63细胞株CLCF1基因的表达，这可能是绝经后骨质疏松症肾阴虚证的分子机制之一。
Abstract:
Objective To explore the effect of 17β-estradiol on CLCF1 gene expression in human osteosarcoma cells.Methods The osteosarcoma cells MG-63 were divided into control group, β-estradiol low, medium and high dose groups.In the control group, 10-3mol/L DMSO was added, and 17β-estradiol was diluted with DMSO, and then added to the low, medium, and high-dose β-estradiol groups.The final concentrations were 10-10, 10-9, and 10-8 mol/L. After 24, 48, and 72 h after intervention with MG-63 cells, real-time fluorescence quantitative PCR was used to detect the expression of CLCF1 mRNA in each group of MG-63 cells;Construction of CLCF1 gene promoter luciferase reporter gene vector, using 10-8 mol/L 17β-estradiol and estrogen receptor inhibitor Fulvestrant (ICI) to intervene in MG63 cells for 24 h, double luciferase reporter gene The system detects dual luciferase activity.Results ① After 24 h of intervention, the expression of CLCF1 mRNA in β-estradiol low, medium and high dose groups was higher than that of the control group, the difference was statistically significant (P<0.05);After 48 h of intervention, only the expression of CLCF1 mRNA in the β-estradiol high-dose group was higher than that of the control group, the difference was statistically significant (P<0.05);After 72 h of intervention, the expression of CLCF1 mRNA in the low β-estradiol group was lower than that of the control group,the difference was statistically significant (P＜0.05).There was no statistically significant difference in the expression of CLCF1 mRNA between the middle and high doses of β-estradiol group and the control group (P>0.05);②Compared with the control group, the luciferase activity of the 17β-estradiol high-dose group increased,the difference was statistically significant(P＜0.05);After adding the estrogen receptor inhibitor ICI, the luciferase activity was lower than the control group,the difference was statistically significant (P<0.05).Conclusion 17β-estradiol can promote the expression of CLCF1 gene in MG63 cell line by increasing the activity of CLCF1 promoter, which may be one of the molecular mechanisms of kidney-yin deficiency syndrome in postmenopausal osteoporosis.

参考文献/References:

[1]陈娟,谢丽华,李生强,等.hsa-miR-655靶向调控绝经后骨质疏松症肾阴虚证关联基因CLCF1的实验研究[J].中国骨质疏松杂志,2016,22(8):939-943. [2]申浩,丁品胜,魏戌,等.基于文献的骨质疏松症中医证候特征研究[J].中国中医基础医学杂志,2018,24(12):1676-1679. [3]Hamilton KJ,Hewitt SC,Arao Y,et al.Estrogen Hormone Biology[J].Curr Top Dev Biol,2017(125):109-146. [4]Xin J,Jing S,Bo Y,et al.Daidzein stimulates osteogenesis facilitating proliferation,differentiation,and antiapoptosis in human osteoblast-like MG-63 cells via estrogen receptor-dependent MEK/ERK and PI3K/Akt activation[J].Nutr Res,2017(42):20-30. [5]Hu B,Sun XW,Yang Y,et al.Tomatidine suppresses osteoclastogenesis and mitigates estrogen deficiency-induced bone mass loss by modulating TRAF6-mediated signaling[J].FASEB J,2019,33(2):2574-2586. [6]沈国蔚,成心锟,颜世昌,等.CFTR在雌激素诱导破骨细胞凋亡中的作用机制研究[J].中国骨与关节损伤杂志,2018,33(11):1150-1152. [7]Kim JW,Marquez CP,Kostyrko K,et al.Antitumor activity of an engineered decoy receptor targeting CLCF1-CNTFR signaling in lung adenocarcinoma[J].Nat Med,2019,25(11):1783-1795. [8]Pasquin S,Chehboun S,Dejda A,et al.Effect of human very low-density lipoproteins on cardiotrophin-like cytokine factor 1(CLCF1)activity[J].Sci Rep,2018,8(1):3990. [9]Ge JR,Xie LH,Chen J,et al.Liuwei Dihuang Pill treats postmenopausal osteoporosis with Shen(Kidney)yin deficiency via Janus kinase/signal transducer and activator of transcription signal pathway by up-regulating cardiotrophin-like cytokine factor 1 expression[J].Chin J Integr Med,2018,24(6):415-422. [10]马铭,任汉强.IL-37通过调控M-CSF和抑制IL-6-JAK2/STAT3信号通路抑制骨质疏松的机制研究[J].中国免疫学杂志,2017,33(10):1552-1556. [11]吴景华,郭佳培,王志刚,等.IL-6-JAK2/STAT3信号途径在骨质疏松症的作用机制研究[J].现代预防医学,2016,43(14):2621-2623. [12]余昕,万启龙,李智,等.AG490抑制Jak2-STAT3信号通路调控骨髓间充质干细胞迁移、矿化及骨缺损愈合的机制研究[J].中华口腔医学杂志,2018,53(5):293-300. [13]Wu W,Fu J,Gu Y,et al.JAK2/STAT3 regulates estrogen-related senescence of bone marrow stem cells[J].J Endocrinol.2020,245(1):141-153. [14]程鑫,万启龙,李祖兵.AG490对成骨细胞和破骨细胞之间相互作用的影响[J].中华口腔医学研究杂志,2017,11(2):73-80.

更新日期/Last Update: 1900-01-01