[1]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,(1):17-22.[doi:DOI:10.3969/j.issn.1000-7369.2022.01.004]
　ZHANG Lin,DENG Guihua,YANG Liang,et al.The mechanism of Picrasidine F regulating the proliferation and apoptosis of cervical cancer HeLa cells through miR-331-3p[J].,2022,(1):17-22.[doi:DOI:10.3969/j.issn.1000-7369.2022.01.004]
点击复制

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
分享到：

《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2022年1期
页码:
17-22
栏目:
基础研究
出版日期:
2022-01-05

文章信息/Info

Title:
The mechanism of Picrasidine F regulating the proliferation and apoptosis of cervical cancer HeLa cells through miR-331-3p
作者:
张 琳1; 邓贵华1; 杨 亮1; 肖大立2
(1.广东省妇幼保健院药学部,广东 广州 510010; 2.广东省妇幼保健院医务科,广东 广州 510010)
Author(s):
ZHANG Lin; DENG Guihua; YANG Liang; XIAO Dali
(Department of Pharmacy,Guangdong Maternity and Child Health Hospital,Guangzhou 510010,China)
关键词:
苦木碱F;  miR-331-3p;  宫颈癌;  增殖;  凋亡;  蛋白酶体活化复合物3
Keywords:
Picrasidine F;  miR-331-3p;  Cervical cancer;  Proliferation;  Apoptosis;  Proteasome activator complex subunit 3
分类号:
R 737.33
DOI:
DOI:10.3969/j.issn.1000-7369.2022.01.004
文献标志码:
A
摘要:
目的:探讨苦木碱F调控miR-331-3p在宫颈癌HeLa细胞增殖及凋亡中的作用及机制。方法:MTT法检测苦木碱F对宫颈癌HeLa细胞增殖的影响; 流式细胞术检测苦木碱F对宫颈癌HeLa细胞凋亡的影响; RT-qPCR检测苦木碱F处理HeLa细胞后miR-331-3p的表达; miR-331-3p mimic或miR-331-3p inhibitor 转染HeLa细胞,经苦木碱F处理后检测HeLa细胞的增殖及凋亡情况; 使用生物信息学和双荧光素酶报告基因实验预测及验证miR-331-3p与蛋白酶体活化复合物3(PSME3)的靶向关系; Western blot检测HeLa细胞中凋亡相关蛋白表达情况。结果:苦木碱F能显著抑制HeLa细胞增殖,促进凋亡,且具有浓度依赖性(P<0.01); 苦木碱F显著上调HeLa细胞中miR-331-3p的表达(P<0.01); miR-331-3p mimic能显著抑制HeLa细胞增殖能力,促进细胞凋亡; miR-331-3p inhibitor能显著降低苦木碱F对HeLa细胞增殖的抑制以及凋亡的促进作用(P<0.05); 生物信息学预测PSME3是miR-331-3p的靶基因之一,双荧光素酶报告基因结果显示miR-331-3p与PSME3 3'UTR靶向结合; 与苦木碱F+NC inhibitor +NC-siRNA组比较,苦木碱F+miR-331-3p inhibitor +NC-siRNA组中细胞存活率升高,细胞凋亡率降低,β-catenin和Bcl-2蛋白水平升高,Bax蛋白水平降低,而敲低PSME3能逆转这一结果,差异有统计学意义(P<0.05)。结论:苦木碱F通过介导miR-331-3p/PSME3轴抑制宫颈癌HeLa细胞增殖,促进凋亡,发挥抗肿瘤作用。
Abstract:
Objective:To investigate the effect of Picrasidine F-mediated miR-331-3p on the proliferation and apoptosis of cervical cancer HeLa cells and its underlying mechanism.Methods:MTT assay was used to detect the effect of Picrasidine F on the proliferation of cervical cancer HeLa cells.The effect of Picrasidine F on apoptosis of cervical carcinoma HeLa cells was detected by flow cytometry.The expression of miR-331-3p in Hela cells treated with Picrasidine F was detected by RT-qPCR.MiR-331-3p mimic or miR-331-3p inhibitor was transfected into HeLa cells,and the proliferation and apoptosis of HeLa cells were detected after treatment with Picrasidine F.Bioinformatics and dual luciferase reporter gene assay were used to predict and verify the targeting relationship between miR-331-3p and proteasome activator complex subunit 3(PSME3).The expression of apoptosis-related protein in HeLa cells was detected by Western blot.Results:Picrasidine F significantly inhibited the proliferation and promoted apoptosis of HeLa cells in a concentration-dependent manner(P<0.01).Picrasidine F significantly up-regulated the expression of miR-331-3p in HeLa cells(P<0.01).MiR-331-3p mimic could significantly inhibit the proliferation of HeLa cells and promote cell apoptosis.MiR-331-3p inhibitor significantly reduced the inhibitory effect of Picrasidine F on HeLa cell proliferation and the promotion of apoptosis(P<0.05).Bioinformatics predicted that PSME3 was one of the target genes of miR-331-3p,and dual luciferase reporter gene results showed that miR-331-3p was targeted to bind PSME3 3'UTR.Compared with the Picrasidine F+NC inhibitor+NC-siRNA group,the survival rate and apoptosis rate of cells in the Picrasidine F+miR-331-3p inhibitor+NC-siRNA group were increased,the protein levels of β-catenin and Bcl-2 were increased,and the protein levels of Bax were decreased,while knockdown of PSME3 could reverse these results,and the difference was statistically significant(P<0.05).Conclusion:Picrasidine F played an anti-tumor role by mediating miR-331-3p/PSME3 axis to inhibit the proliferation and promote apoptosis of cervical cancer HeLa cells.

参考文献/References:

[1] 崔开颖,朱根海.低分子肝素对宫颈癌细胞增殖及凋亡的影响[J].中国临床药理学杂志,2020,32(18):108-112.
[2] 郝志晔,张 晶,钱 伟,等.参芪排毒汤对宫颈癌组织人乳头瘤病毒E6/E7、P53和PRb基因蛋白表达的影响[J].陕西中医,2021,42(2):192-195,199.
[3] 刘 岩,张 虹,戴 玮,等.苦木对HepG-2细胞增殖抑制作用及机制的研究[J].中药材,2010,33(7):1143-1146.
[4] Chen Z,Zhang M,Qiao Y,et al.MicroRNA-1297 contributes to the progression of human cervical carcinoma through PTEN[J].Artif Cells Nanomed Biotechnol,2018,46(s2):1120-1126.
[5] 朱文侠,杨 玲,成 钧,等.miR-142-3p在结直肠癌组织与细胞中的表达及对细胞增殖影响研究[J].陕西医学杂志,2018,47(3):282-286.
[6] Yang L,Liu L,Zhang X,et al.miR-96 enhances the proliferation of cervical cancer cells by targeting FOXO1[J].Pathol Res Pract,2020,216(4):152854.
[7] 赵 敏,赵 琳,钟社兰.MiR-123调控Bcl-2对宫颈癌细胞侵袭及转移能力的影响[J].陕西医学杂志,2020,49(5):542-544,548.
[8] Tomomi F,Keiji S,Aya A,et al.MicroRNA-331-3p Suppresses cervical cancer cell proliferation and E6/E7 Expression by targeting NRP2[J].International Journal of Molecular Sciences,2016,17(8):1351.
[9] Song L,Liu D,Zhao Y,et al.Sinomenine inhibits breast cancer cell invasion and migration by suppressing NF-kappaB activation mediated by IL-4/miR-324-5p/CUEDC2 axis[J].Biochem Biophys Res Commun,2015,464(3):705-710.
[10] Koike K,Ohmoto T,Ogata K.Studies on the alkaloids of picrasmaqu assioidesbennet viii:X-ray crystal structure analysis of picrasidine-f[J].Chem Pharm Bull,1986,34(8):3228-3236.
[11] Yamashita N,Kondo M,Zhao S,et al.Picrasidine G decreases viability of MDA-MB 468 EGFR-overexpressing triple-negative breast cancer cells through inhibition of EGFR/STAT3 signaling pathway[J].Bioorg Med Chem Lett,2017,27(11):2608-2612.
[12] 石国华,焦伟华,杨 帆,等.苦木中3个二聚β-卡巴林生物碱及其生物活性研究[J].中药材,2015,46(6):803-807.
[13] 朱 亚,张 伟,柯丽娜,等.miRNA-41调控c-myc在宫颈癌上皮间质转化中的作用[J].河北医药,2020,42(1):113-115,119.
[14] Yin XZ,Zhao DM,Zhang GX,et al.Effect of miRNA-203 on cervical cancer cells and its underlying mechanism[J].Genet Mol Res,2016,15(3):8680.
[15] 莫鸿英,李玉飞.miRNA221对宫颈癌细胞株Siha侵袭转移行为的影响[J].湖南师范大学学报:医学版,2017,14(3):63-66.
[16] 陈国栋,文 武,贺 军,等.miR-331-3p在肝细胞癌中表达的研究[J].中国当代医药,2014,7(17):4-7.
[17] 李 静.miR-331-3p在恶性肿瘤中的研究进展[J].实用肿瘤学杂志,2018,32(2):125-129.
[18] 王 伟,陈传好.microRNA在肿瘤治疗中的研究进展[J].基因组学与应用生物学,2017,36(7):2804-2809.
[19] 白 雪,张 斌,汪 潇,等.miR-637调控RING1表达抑制口腔鳞癌Tac-8113细胞增殖实验研究[J].陕西医学杂志,2020,49(8):923-927.
[20] Zhao D,Sui Y,Zheng X.miR-331-3p inhibits proliferation and promotes apoptosis by targeting HER2 through the PI3K/Akt and ERK1/2 pathways in colorectal cancer[J].Oncol Rep,2016,35(2):1075-1082.
[21] Tian QQ,Xia J,Zhang X,et al.miR-331-3p inhibits tumor cell proliferation,metastasis,invasion by targeting mllt10 in non-small cell lung cancer[J].Cancer Manag Res,2020,12:5749-5758.
[22] Shi Y,Luo X,Li P,et al.miR-7-5p suppresses cellproliferation and induces apoptosis of breast cancer cells mainly by targetingREGgamma[J].Cancer Lett,2015,358:e36.
[23] Wang H,Bao W,Jiang F,et al.Mutant p53(p53-R248Q)functions as an oncogene in promoting endometrial cancer by upregulating REG gamma[J].Cancer Lett,2015,36:e279.
[24] 向秋玲,李雪兰,黄薇霖,等.槲皮素通过G3BP1调控Wnt信号通路抑制宫颈癌细胞增殖[J].中药药理与临床,2020,36(4):102-108.

备注/Memo

备注/Memo:
基金项目:广东省中医药局科研项目(20181038)
更新日期/Last Update: 2022-01-09