[1]王京京,李全福,张立广,等.紫檀芪调控微小RNA-340-5p/髓细胞白血病-1轴对胆囊癌SD细胞生物学行为的影响实验研究[J].陕西医学杂志,2024,(3):322-326,330.[doi:DOI:10.3969/j.issn.1000-7377.2024.03.007]
　WANG Jingjing,LI Quanfu,ZHANG Liguang,et al.Effects of pterostilbene regulating miR-340-5p/MCL-1 axis on biological behavior of gallbladder cancer GBC-SD cells[J].,2024,(3):322-326,330.[doi:DOI:10.3969/j.issn.1000-7377.2024.03.007]
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紫檀芪调控微小RNA-340-5p/髓细胞白血病-1轴对胆囊癌SD细胞生物学行为的影响实验研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
期数:
2024年3期
页码:
322-326,330
栏目:
基础研究
出版日期:
2024-03-05

文章信息/Info

Title:
Effects of pterostilbene regulating miR-340-5p/MCL-1 axis on biological behavior of gallbladder cancer GBC-SD cells
作者:
王京京; 李全福; 张立广; 邵青龙
(保定市第二医院肝胆外科,河北 保定 071000)
Author(s):
WANG Jingjing; LI Quanfu; ZHANG Liguang; SHAO Qinglong
(Department of Hepatobiliary Surgery,the No.2 Hospital of Baoding,Baoding 071000,China)
关键词:
胆囊癌;  紫檀芪;  微小RNA-340-5p;  髓细胞白血病-1;  GBC-SD细胞;  生物学行为
Keywords:
Gallbladder cancer;  Pterostilbene;  miR-340-5p;  Myeloid cell leukemia-1;  GBC-SD cells;  Biological behavior
分类号:
R 285
DOI:
DOI:10.3969/j.issn.1000-7377.2024.03.007
文献标志码:
A
摘要:
目的:探究紫檀芪(PTS)调控微小RNA-340-5p(miR-340-5p)/髓细胞白血病-1(MCL-1)轴对胆囊癌SD(GBC-SD)细胞增殖、凋亡、迁移和侵袭的影响。方法:体外培养人GBC-SD细胞,使用不同浓度PTS(0、5、10、20、40、80 μmol/L)处理GBC-SD细胞,然后检测细胞存活率。将GBC-SD细胞分为对照组(DMEM培养基常规培养GBC-SD细胞)、PTS组(加入80 μmol/L的PTS培养GBC-SD细胞)、PTS+空载质粒组(加入80 μmol/L的PTS培养GBC-SD细胞,并转染空载质粒)、PTS+miR-340-5p沉默组[加入80 μmol/L的PTS培养GBC-SD细胞,并转染miR-340-5p小干扰RNA(siRNA)质粒]。分别检测各组GBC-SD细胞增殖、凋亡、迁移和侵袭能力。实时荧光定量PCR(RT-qPCR)检测miR-340-5p和MCL-1 mRNA表达。蛋白印迹法检测MCL-1及增殖凋亡相关蛋白表达。双荧光素酶报告基因实验检测miR-340-5p与MCL-1的靶向关系。裸鼠移植瘤实验检测PTS对裸鼠肿瘤生长及miR-340-5p/MCL-1轴的影响。结果:随着PTS浓度的升高,GBC-SD细胞存活率逐渐降低(均P<0.05)。与对照组比较,PTS组细胞存活率、集落形成数、细胞迁移率、细胞侵袭数、MCL-1 mRNA和蛋白、Ki67、细胞周期蛋白D1(Cyclin D1)、B淋巴细胞瘤-2(Bcl-2)蛋白表达降低,细胞凋亡率、miR-340-5p、Bcl-2相关X蛋白(Bax)、活化的半胱氨酸天冬氨酸蛋白酶-3(Cleaved caspase-3)蛋白表达升高(均P<0.05)。沉默miR-340-5p可逆转PTS对GBC-SD细胞增殖、凋亡、迁移、侵袭及miR-340-5p/MCL-1轴的作用效果(均P<0.05)。miR-340-5p与MCL-1存在靶向调控关系。PTS能抑制裸鼠肿瘤生长和MCL-1 mRNA和蛋白表达,促进miR-340-5p表达(均P<0.05)。结论:PTS可通过上调miR-340-5p表达下调MCL-1表达,抑制GBC-SD细胞增殖、迁移和侵袭,促进GBC-SD细胞凋亡。
Abstract:
Objective:To explore the effects of pterostilbene(PTS)regulating microRNA-340-5p(miR-340-5p)/myeloid cell leukemia-1(MCL-1)axis on the proliferation,apoptosis,migration and invasion of GBC-SD cells in gallbladder cancer(GBC).Methods:Human GBC-SD cells were cultured in vitro and treated with different concentrations of PTS(0,5,10,20,40,80 μmol/L)to detect the cell survival rate.GBC-SD cells were divided into control group(conventional culture of GBC-SD cells in DMEM medium),PTS group(adding 80 μmol/L PTS to culture GBC-SD cells),PTS+unloaded plasmid group(adding 80 μmol/L PTS to culture GBC-SD cells,and transfected empty plasmid),PTS+miR-340-5p silencing group(80 μmol/L PTS were added to culture GBC-SD cells and transfected miR-340-5p siRNA plasmid).The proliferation,apoptosis,migration and invasion of GBC-SD cells in each group were detected.The expressions of miR-340-5p and MCL-1 mRNA were detected by RT-qPCR.The expressions of MCL-1 and proliferation-apoptosis related proteins were detected by Western blot.Targeting relationship between miR-340-5p and MCL-1 was detected by dual luciferase reporter gene detection.The effects of PTS on tumor growth and miR-340-5p/MCL-1 axis in nude mice were detected by tumor transplantation experiment.Results:With the increase of PTS concentration,the survival rate of BGC-SD cells decreased gradually(all P<0.05).Compared with control group,the cell survival rate,colony formation number,mobility,cell invasion number,MCL-1 mRNA and protein,Ki67,Cyclin D1 and Bcl-2 protein expressions in PTS group were significantly decreased,apoptosis rate,miR-340-5p,Bax,Cleaved caspase-3 protein expressions were significantly increased(all P<0.05).Silencing miR-340-5p could reverse the effects of PTS on the proliferation,apoptosis,migration,invasion and miR-340-5p/MCL-1 axis of BGC-SD cells(all P<0.05).MiR-340-5p has a targeted regulatory relationship with MCL-1.PTS could inhibit tumor growth,mRNA and protein expression of MCL-1,and promote the expression of miR-340-5p in nude mice(all P<0.05).Conclusion:PTS can upregulate the expression of miR-340-5p,and then down-regulate the expression of MCL-1,inhibit the proliferation,migration and invasion of BGC-SD cells,and promoting GBC-SD cell apoptosis.

参考文献/References:

[1] ROA J C,GARCIA P,KAPOOR V K,et al.Gallbladder cancer[J].Nat Rev Dis Primers,2022,8(1):69-75.
[2] 田刚,魏梦绮,王霞,等.CT检查在黄色肉芽肿性胆囊炎与胆囊癌鉴别诊断中的价值[J].陕西医学杂志,2022,51(12):1528-1531.
[3] LV Y,YIN W,ZHANG Z.Non-coding RNAs as potential biomarkers of gallbladder cancer[J].Clin Transl Oncol,2023,25(6):1489-1511.
[4] 姚雷,蔡海建,刘邓,等.紫檀芪减轻小鼠肝窦内皮细胞氧化损伤的机制[J].中国药理学通报,2023,39(2):310-314.
[5] 付华君,宋文英,田俊斌,等.紫檀芪通过调控细胞自噬对抗大鼠心肌缺血/再灌注损伤[J].陕西中医,2022,43(10):1342-1346.
[6] WAWSZCZYK J,JESSE K,KAPRAL M.Pterostilbene-mediated inhibition of cell proliferation and cell death induction in amelanotic and melanotic melanoma[J].Int J Mol Sci,2023,24(2):1115-1133.
[7] WANG Z,WANG T,CHEN X,et al.Pterostilbene regulates cell proliferation and apoptosis in non-small-cell lung cancer via targeting COX-2[J].Biotechnol Appl Biochem,2023,70(1):106-119.
[8] GAO H,LIU Z,XU W,et al.Pterostilbene promotes mitochondrial apoptosis and inhibits proliferation in glioma cells[J].Sci Rep,2021,11(1):6381-6398.
[9] TONG C,WANG Y,LI J,et al.Pterostilbene inhibits gallbladder cancer progression by suppressing the PI3K/Akt pathway[J].Sci Rep,2021,11(1):4391-4405.
[10] CAO J,SHAO H,HU J,et al.Identification of invasion-metastasis associated miRNAs in gallbladder cancer by bioinformatics and experimental validation[J].J Transl Med,2022,20(1):188-201.
[11] LIU L,YAN Y,ZHANG G,et al.Knockdown of LINC01694 inhibits growth of gallbladder cancer cells via miR-340-5p/Sox4[J].Biosci Rep,2020,40(4):4444-4455.
[12] YU X,ZHOU L,LIU W,et al.Skp2 stabilizes Mcl-1 and confers radioresistance in colorectal cancer[J].Cell Death Dis,2022,13(3):249-261.
[13] WANG S,SU T T,TONG H,et al.CircPVT1 promotes gallbladder cancer growth by sponging miR-339-3p and regulates MCL-1 expression[J].Cell Death Discov,2021,7(1):191-204.
[14] 王若利,王潇,岳红梅.紫檀芪对肺纤维化小鼠的抗氧化作用研究[J].中国临床药理学杂志,2021,37(5):532-535.
[15] HEMANI R,PATEL I,INAMDAR N,et al.Dietary pterostilbene for MTA1-targeted interception in high-risk premalignant prostate cancer[J].Cancer Prev Res(Phila),2022,15(2):87-100.
[16] HOJO Y,KISHI S,MORI S,et al.Sunitinib and pterostilbene combination treatment exerts antitumor effects in gastric cancer via suppression of PDZD8[J].Int J Mol Sci,2022,23(7):4002-4019.
[17] WAWSZCZYK J,JESSE K,SMOLIK S,et al.Mechanism of pterostilbene-induced cell death in HT-29 colon cancer cells[J].Molecules,2022,27(2):369-387.
[18] YOULIN K,SIMIN L,JIAN K,et al.Inhibition of miR-20a by pterostilbene facilitates prostate cancer cells killed by NK cells via up-regulation of NKG2D ligands and TGF-β1 down-regulation[J].Heliyon,2023,9(4):14957-14967.
[19] 张琳,邓贵华,杨亮,等.苦木碱F通过miR-331-3p调控宫颈癌HeLa细胞增殖及凋亡的机制研究[J].陕西中医,2022,43(1):17-22.
[20] 依力哈木?买买提,钟锴,艾合散?卡马力,等.环状RNA circ_0020123靶向调控微小RNA-145影响胰腺癌细胞SW-1990增殖、侵袭与转移实验研究[J].陕西医学杂志,2023,52(9):1140-1144.
[21] ZENG F,LUO L,SONG M,et al.Silencing of circular RNA PUM1 inhibits clear cell renal cell carcinoma progression through the miR-340-5p/FABP7 axis[J].J Recept Signal Transduct Res,2022,42(2):141-150.
[22] YANG K D,WANG Y,ZHANG F,et al.CircN4BP2L2 promotes colorectal cancer growth and metastasis through regulation of the miR-340-5p/CXCR4 axis[J].Lab Invest,2022,102(1):38-47.
[23] KHAN S,KELLISH P,CONNIS N,et al.Co-targeting BCL-XL and MCL-1 with DT2216 and AZD8055 synergistically inhibit small-cell lung cancer growth without causing on-target toxicities in mice[J].Cell Death Discov,2023,9(1):1-9.
[24] HSIEH Y H,YU F J,NASSEF Y,et al.Targeting of MCL-1 expression by miRNA-3614-5p promotes cell apoptosis of human prostate cancer cells[J].Int J Mol Sci,2022,23(8):4194-4206.
[25] WINDER M L,CAMPBELL K J.MCL-1 is a clinically targetable vulnerability in breast cancer[J].Cell Cycle,2022,21(14):1439-1455.

备注/Memo

备注/Memo:
基金项目:河北省医学科学研究课题(20220260); 河北省保定市科技计划项目(2041ZF213)
更新日期/Last Update: 2024-03-05