[1]王敬贤,靳宪辉,庞巨涛,等.槟榔碱通过断裂转导蛋白样增强子4对裸鼠骨肉瘤细胞自噬和凋亡的影响实验研究[J].陕西医学杂志,2024,(12):1611-1616,1623.[doi:DOI:10.3969/j.issn.1000-7377.2024.12.005]
　WANG Jingxian,JIN Xianhui,PANG Jutao,et al.Effect of arecoline on autophagy and apoptosis in nude mice osteosarcoma cells via TLE4[J].,2024,(12):1611-1616,1623.[doi:DOI:10.3969/j.issn.1000-7377.2024.12.005]
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槟榔碱通过断裂转导蛋白样增强子4对裸鼠骨肉瘤细胞自噬和凋亡的影响实验研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
期数:
2024年12期
页码:
1611-1616,1623
栏目:
基础研究
出版日期:
2024-12-05

文章信息/Info

Title:
Effect of arecoline on autophagy and apoptosis in nude mice osteosarcoma cells via TLE4
作者:
王敬贤1; 靳宪辉2; 庞巨涛1; 张 伟1
(1.衡水市人民医院关节骨科运动医学病区,河北 衡水 053000; 2.衡水市人民医院脊柱骨科骨肿瘤病区,河北 衡水 053000)
Author(s):
WANG Jingxian; JIN Xianhui; PANG Jutao; ZHANG Wei
(Sports Medicine Ward,Department of Joint Orthopedics,Hengshui People's Hospital,Hengshui 053000,China)
关键词:
槟榔碱;  骨肉瘤;  断裂转导蛋白样增强子4;  自噬;  凋亡;  移植瘤;  裸鼠
Keywords:
Arecoline;  Osteosarcoma;  Transducin-like enhancer of split 4;  Autophagy;  Apoptosis;  Transplanted tumor;  Nude mice
分类号:
R 285
DOI:
DOI:10.3969/j.issn.1000-7377.2024.12.005
文献标志码:
A
摘要:
目的:探究槟榔碱通过断裂转导蛋白样增强子4(TLE4)对大鼠骨肉瘤细胞自噬和凋亡的影响。方法:选取SPF级裸鼠建立骨肉瘤移植瘤模型。将建模成功的60只裸鼠随机分为模型组,槟榔碱低、中、高剂量组(分别腹腔注射10、20、50 mg/kg槟榔碱),TLE4 抑制剂组(腹腔注射3 mg/kg TAK-242)和顺铂组(腹腔注射2 mg/kg顺铂),每组10只,隔日注射1次,连续注射14 d。将未建模的10只裸鼠作为对照组。对照组和模型组每天注射同体积0.9%氯化钠溶液。观察移植瘤体积,取瘤体进行称重。实时荧光定量PCR(RT-PCR)检测骨肉瘤组织TLE4 mRNA表达,免疫组织化学染色检测病理学变化和TLE4蛋白表达。Western blot检测重组人自噬效应蛋白(Beclin-1)、微管相关蛋白1轻链3Ⅰ(LC3Ⅰ)、微管相关蛋白1轻链3Ⅱ(LC3Ⅱ)、家蚕隔离体蛋白1(p62)、B淋巴细胞瘤-2(Bcl-2)、Bcl相关X蛋白(Bax)、半胱氨酸蛋白酶-3(Caspase-3)蛋白表达。结果:与模型组比较,槟榔碱低、中、高剂量组以及TLE4 抑制剂组和顺铂组TLE4、移植瘤体积和重量、Beclin-1、LC3Ⅱ/LC3Ⅰ、LC3Ⅱ、Bcl-2降低,Bax、Caspase-3、LC3Ⅰ升高(均P<0.05)。与槟榔碱低剂量组比较,槟榔碱中、高剂量组以及TLE4 抑制剂组和顺铂组TLE4、移植瘤体积和重量、Beclin-1、LC3Ⅱ/LC3Ⅰ、LC3Ⅱ、Bcl-2降低,Bax、Caspase-3、P62、LC3Ⅰ升高(均P<0.05)。与槟榔碱中剂量组比较,槟榔碱高剂量组和TLE4 抑制剂组TLE4、移植瘤体积和重量、LC3Ⅱ/LC3Ⅰ、Bcl-2降低,Bax、P62、LC3Ⅰ升高(均P<0.05)。与槟榔碱高剂量组比较,TLE4 抑制剂组和顺铂组Beclin-1、LC3Ⅱ/LC3Ⅰ、LC3Ⅱ升高,P62、LC3Ⅰ、Bax、Caspase-3降低(均P<0.05)。TLE4 抑制剂组和顺铂组以上指标比较差异无统计学意义(均P>0.05)。结论:槟榔碱可能通过抑制 TLE4表达影响骨肉瘤细胞自噬和凋亡,从而抑制骨肉瘤的生长。
Abstract:
Objective:To investigate the effect of arecoline on autophagy and apoptosis in nude mice osteosarcoma cells via transducin-like enhancer of split 4(TLE4).Methods:Nude mouse xenograft osteosarcoma models were established using SPF-grade nude mice.Sixty successful model mice were randomly divided into model group,low-,medium-,and high-dose arecoline groups(intraperitoneal injection of 10,20,and 50 mg/kg arecoline,respectively),TLE4 inhibitor group(intraperitoneal injection of 3 mg/kg TAK-242),and cisplatin group(intraperitoneal injection of 2 mg/kg cisplatin).Each group contained 10 mice.Injections were given every other day for a total of 14 days.Ten unmodeled mice served as the control group.The control group and the model group received daily injections of an equal volume of 0.9% sodium chloride solution.The tumor volumes were observed,and the tumors were weighed at the end of the experiment.RT-qPCR was used to detect the expression of TLE4 mRNA in osteosarcoma tissue.Immunohistochemistry was used to examine the histopathological changes and TLE4 expression in the tissues.Western blot was employed to detect the protein expression levels of LC3Ⅰ,LC3Ⅱ,p62,Bax,Bcl-2,and Caspase-3.Results:Compared with the model group,the TLE4,tumor volume and weight,Beclin-1,LC3Ⅱ/LC3Ⅰ ratio,LC3Ⅱ,and Bcl-2 levels were significantly reduced,while the levels of Bax,Caspase-3,and LC3Ⅰ were significantly increased in the low-,medium-,and high-dose arecoline groups,the TLE4 inhibitor group,and the cisplatin group(all P<0.05).Compared with the low-dose arecoline group,the TLE4,tumor volume and weight,Beclin-1,LC3Ⅱ/LC3Ⅰ ratio,LC3Ⅱ,and Bcl-2 levels were significantly reduced,while the levels of Bax,Caspase-3,p62,and LC3Ⅰ were significantly increased in the medium- and high-dose arecoline groups,the TLE4 inhibitor group,and the cisplatin group(all P<0.05).Compared with the medium-dose arecoline group,the TLE4,tumor volume and weight,LC3Ⅱ/LC3Ⅰ ratio and Bcl-2 levels were significantly reduced,while the levels of Bax、P62、LC3Ⅰ were significantly increased in the high-dose arecoline group and the TLE4 inhibitor group(all P<0.05).Compared with the high-dose arecoline group,the Beclin-1、LC3Ⅱ/LC3Ⅰ、LC3Ⅱ levels were significantly increased,whereas the levels of P62、LC3Ⅰ、Bax、Caspase-3 were significantly decreased in the TLE4 inhibitor group and the Cisplatin group(all P<0.05).There was no significant difference between the TLE4 inhibitor group and the cisplatin group regarding these indicators(all P>0.05).Conclusion:Arecoline can inhibit the growth of osteosarcoma,and its mechanism may involve suppressing the expression of TLE4 in osteosarcoma cells,thereby regulating autophagy and apoptosis.

参考文献/References:

[1] LIU Y,GUO Y,WANG Z,et al.Current status of osteosarcoma therapy:Challenges and opportunities[J].Molecular Cancer,2020,19(1):1-14.
[2] 张治业,余醒醒,张璐,等.CD166对多发性骨髓瘤RPMI-8226细胞增殖、凋亡及自噬的影响实验研究[J].陕西医学杂志,2023,52(12):1675-1679.
[3] ELMORE S.Apoptosis:A review of programmed cell death[J].Toxicol Pathol,2007,35(4):495-516.
[4] LI X,WU H,ZHANG L,et al.TLE4 regulates autophagy and cell survival by interacting with Beclin1 in colorectal cancer[J].Cell Death Dis,2019,10(1):1-13.
[5] ZHOU J,ZHANG J,CHEN Q,et al.TLE4 promotes cell proliferation and inhibits apoptosis in gastric cancer by regulating miR-195-5p/CCND1 axis[J].Cancer Lett,2021,504:164-174.
[6] WANG H,ZHANG Y,LI Z,et al.Arecoline inhibits growth and induces apoptosis in human lung cancer cells via the ROS/JNK/p53 signaling pathway[J].J Cell Physiol,2019,234(12):22097-22108.
[7] 邓国磊,何双桃,李国良,等.槟榔碱通过氧化应激介导的内皮间充质转化致HUVE细胞损伤的机制研究[J].肿瘤药学,2022,12(4):488-495.
[8] 李静.氢溴酸槟榔碱调控肝癌Hep3B细胞增殖和凋亡的功能研究[D].重庆:重庆大学,2020.
[9] 靳婷.槟榔提取物抗高原缺氧药效学及其机制研究[D].兰州:兰州大学,2018.
[10] 陈敏,刘晓梅,魏玲格,等.99Tcm-MAG3-VEGF125-136的制备及在骨肉瘤荷瘤裸鼠模型中的生物分布与显像[J].中华核医学与分子影像杂志,2016,36(5):450-453.
[11] 胡晨,张昊芃,张晓朦,等.槟榔碱细胞毒性及其作用机制研究进展[J].中成药,2023,45(10):3364-3368.
[12] 朱冰玉,蒋琪.槟榔碱对口腔黏膜上皮细胞自噬和凋亡的影响[J].现代实用医学,2023,35(7):866-869.
[13] 唐乃高,郑根建.miR-486-3p靶向DDR1对口腔鳞状细胞癌的抑制作用及槟榔致口腔鳞状细胞癌的可能机制[J].广西医学,2022,44(20):2384-2390.
[14] 贾哲,韩婷,刘欢,等.基于多元统计分析的食用槟榔及药用槟榔主要化学成分的含量对比研究[J].中华中医药杂志,2017,32(11):5158-5161.
[15] LIU Y,ZHANG Y,WANG X,et al.Investigating the cytotoxic effects of arecoline on osteosarcoma cells[J].Cancer Sci,2020,5(3):231-237.
[16] LIU H,ZHAO Y,CHEN M,et al.Comprehensive overview of arecoline's molecular targets in cancer treatment strategies[J].Pharmacol Res,2019,142:133-141.
[17] WANG L,ZHANG J,LIU Y,et al.TLE4:Emerging roles in oncogenesis and tumor progression[J].Cancer Lett,2018,424:1-9.
[18] XIE M,ZHANG J,WANG Y,et al.TLE4 inhibition suppresses tumor growth by regulating autophagy and apoptosis in osteosarcoma[J].J Cancer,2020,11(12):3493-3503.
[19] RONG Y,FENG F Y,GAO W,et al.Autophagy regulation in cancer metabolism and therapy resistance[J].Signal Transduc Tar,2021,6(1):1-12.
[20] 杨竣然,徐子文,陈遒,等.补阳还五汤有效组分抗缺血性脑卒中研究进展[J].陕西中医,2024,45(7):995-998.
[21] MAIURI M C,GALLUZZI L,ZALCKVAR E,et al.Self-eating and self-killing:Crosstalk between autophagy and apoptosis[J].Nat Rev Mol Cell Bio,2018,19(10):657-674.
[22] GANLEY I G,JIANG X,NEUFELD T P,et al.Autophagy and its relation to cell death[J].Annu Rev Cell Dev Bio,2017,33:233-256.
[23] KLIONSKY D J,ABDELMOHSEN K,ABE A,et al.Guidelines for the use and interpretation of assays for monitoring autophagy(5th edition)[J].Autophagy,2021,17(1):1-382.
[24] LI C,SUN L,WANG X,et al.Natural products as potential regulators of autophagy and apoptosis in cancer therapy[J].Pharmacol Res,2019,145:353-362.
[25] CHAUDHRY G E,MDAKIM A,SUNG Y Y,et al.Cancer and apoptosis:The apoptotic activity of plant and marine natural products and their potential as targeted cancer therapeutics[J].Front Pharmacol,2022,13:842376.

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备注/Memo

备注/Memo:
基金项目:河北省医学科学研究课题(20210322,20191769)
更新日期/Last Update: 2024-12-05