[1]张鹏飞,刘新奇,石 菲.miR-35对胃癌细胞MNK-45凋亡的作用及机制探讨[J].陕西医学杂志,2022,51(2):239-244.[doi:DOI:10.3969/j.issn.1000-7377.2022.02.026]
　ZHANG Pengfei,LIU Xinqi,SHI Fei.Effect and mechanism of miR-35 on apoptosis of gastric cancer cell line MNK-45[J].,2022,51(2):239-244.[doi:DOI:10.3969/j.issn.1000-7377.2022.02.026]
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miR-35对胃癌细胞MNK-45凋亡的作用及机制探讨
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
51
期数:
2022年2期
页码:
239-244
栏目:
临床病理
出版日期:
2022-02-05

文章信息/Info

Title:
Effect and mechanism of miR-35 on apoptosis of gastric cancer cell line MNK-45
作者:
张鹏飞1; 刘新奇1; 石 菲2
(1.榆林市第一医院普外科,陕西 榆林 719000; 2.空军军医大学航空航天医学院,陕西 西安 710032)
Author(s):
ZHANG Pengfei; LIU Xinqi; SHI Fei
(Department of General Surgery,the First Hospital of Yulin,Yulin 719000,China)
关键词:
胃癌;  微小RNA;  miR-35;  凋亡;  凋亡通路;  Ras同源基因家族成员A
Keywords:
Gastric cancer;  microRNA;  miR-35;  Apoptosis;  Apoptosis signaling pathway;  Ras homolog gene family member A
分类号:
R 735.2
DOI:
DOI:10.3969/j.issn.1000-7377.2022.02.026
文献标志码:
A
摘要:
目的:探究miR-35对胃癌细胞MNK-45凋亡的作用及机制。方法:利用q-PCR技术检测胃癌细胞系MNK-45中miR-35的含量; 利用细胞转染方法分别将miR-35的阴性对照序列(对照组)、miR-35的模拟序列(模拟物组)或miR-35的互补配对序列(抑制剂组)转染入胃癌细胞MNK-45后,利用荧光显像技术和q-PCR技术检测转染效果; 利用流式细胞仪检测转染后各组细胞系凋亡水平,再进一步利用形态学手段评估细胞凋亡情况; 利用Western blot技术检测凋亡相关蛋白相对含量; 利用荧光素酶报告基因技术检测miR-35的下游靶基因。结果:多种胃癌细胞系中miR-35的表达较正常胃黏膜细胞低(P<0.05); 与对照组相比,转染miR-35模拟序列后MNK-45内miR-35含量增加(P<0.05),转染miR-35的互补配对序列后MNK-45内miR-35含量降低(P<0.05); 与对照组相比,转染miR-35模拟序列后MNK-45细胞凋亡增加(P<0.05),转染miR-35的互补配对序列后MNK-45细胞凋亡减少(P<0.05); 与对照组相比,转染miR-35模拟序列后MNK-45内Caspase 3和Bax等促凋亡蛋白表达增加(均P<0.05),而Bcl-2表达减少(P<0.05); 转染miR-35的互补配对序列后MNK-45细胞内Caspase 3和Bax等促凋亡蛋白表达减少,而Bcl-2表达增加(均P<0.05); 与对照组相比,miR-35模拟序列抑制Ras同源基因家族成员A(RhoA)3'UTR质粒的荧光素酶活性(P<0.05); 与对照组相比,miR-35模拟序列抑制RhoA蛋白表达(P<0.05),而miR-35的互补配对序列促进RhoA蛋白表达(P<0.05)。结论:胃癌细胞系内低表达miR-35,miR-35可通过靶向RhoA促进胃癌细胞MNK-45的凋亡。
Abstract:
Objective:To explore the effect and mechanism of miR-35 on the apoptosis of gastric cancer cell line MNK-45.Methods:The content of miR-35 in gastric cancer cell line MNK-45 was detected by q-PCR.The negative control sequence of miR-35(control group),mimetic sequence of miR-35(mimetic group)or complementary paired sequence of miR-35(inhibitor group)were transfected into gastric cancer cell MNK-45 by cell transfection method.The transfection effect was detected by fluorescence imaging and q-PCR.Flow cytometry and morphological assessment were used to detect the cell apoptosis.The relative content of apoptosis related proteins was detected by Western blot.Luciferase reporter gene technology was used to detect the downstream target gene of miR-35.Results:Compared with normal gastric mucosa cells,miR-35 was decreased in many gastric cancer cell lines(P<0.05).Compared with the control group,miR-35 mimics increased the level of miR-35 while miR-35 inhibitor decreased the level of miR-35 in MNK-45(all P<0.05).Compared with the control group,miR-35 mimics promoted the apoptosis of MNK-45,while miR-35 inhibitor inhibited the apoptosis of MNK-45(all P<0.05).Compared with the control group,miR-35 mimics could increase the expression of apoptotic proteins such as Caspase 3 and Bax,and reduce the expression of Bcl-2,while miR-35 inhibitor reduce the expression of apoptotic proteins and increase the expression of Bcl-2(all P<0.05).Compared with the control group,miR-35 mimics inhibited the luciferase activity of Ras homolog gene family member A(RhoA)3'UTR plasmid(P<0.05).Compared with the control group,miR-35 mimics inhibited the expression of RhoA protein,while miR-35 inhibitor increase the expression of RhoA protein(P<0.05).Conclusion:The level of miR-35 is decreased in gastric cancer cell lines,and miR-35 may promote apoptosis of gastric cancer cell line MNK-45 by targeting RhoA.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金资助项目(31971171)
更新日期/Last Update: 2022-02-09