[1]郝艳芳,刘亚荣,黄玲玲,等.miRNA-195靶向调控 CCND2和 MYB抑制宫颈癌细胞增殖、迁移的机制研究[J].现代检验医学杂志,2022,37(01):130-135.[doi:10.3969/j.issn.1671-7414.2022.01.026]
　HAO Yan-fang,LIU Ya-rong,HUANG Ling-ling,et al.Study on the Mechanism of miRNA-195 Targeting CCND2 and MYB to Inhibit the Proliferation and Migration of Cervical Cancer Cells[J].Journal of Modern Laboratory Medicine,2022,37(01):130-135.[doi:10.3969/j.issn.1671-7414.2022.01.026]
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miRNA-195靶向调控 CCND2和 MYB抑制宫颈癌细胞增殖、迁移的机制研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第37卷
期数:
2022年01期
页码:
130-135
栏目:
论　著
出版日期:
2022-01-15

文章信息/Info

Title:
Study on the Mechanism of miRNA-195 Targeting CCND2 and MYB to Inhibit the Proliferation and Migration of Cervical Cancer Cells
文章编号:
1671-7414（2022）01-130-07
作者:
郝艳芳; 刘亚荣; 黄玲玲; 张　源
（河北北方学院附属第一医院妇科，河北张家口 075000）
Author(s):
HAO Yan-fang;  LIU Ya-rong;  HUANG Ling-ling;  ZHANG Yuan
（Department of Gynecology, the First Affiliated Hospital of Hebei North University,Heibei Zhangjiakou 075000, China）
关键词:
微小核糖核酸 -195; 细胞周期素 D2重组蛋白 2; MYB; 宫颈癌; 增殖; 迁移
分类号:
R737.33；R730.43
DOI:
10.3969/j.issn.1671-7414.2022.01.026
文献标志码:
A
摘要:
目的　研究 miRNA-195在宫颈癌组织和细胞中的表达，分析其对宫颈癌细胞增殖、迁移的影响和作用机制。方法　检测 35例临床宫颈癌组织及其对应癌旁正常组织中 miRNA-195，通过 Kaplan-Meier Plotter数据库分析其与宫颈癌患者预后的关系；采用细胞增殖实验和划痕迁移实验验证 miRNA-195对宫颈癌 siHa，Hela细胞增殖、迁移的影响；通过 microRNA数据库预测 miRNA-195的靶基因，双荧光素酶基因实验验证靶向结合关系； qRT-PCR验证 miRNA-195对靶基因的调控；分析宫颈癌中靶基因的表达作用及与 miRNA-195的相关性，通过细胞回补实验验证 miRNA-195是否通过靶向调控蛋白表达在宫颈癌中发挥功能。结果　宫颈癌组织中 miRNA-195相对表达低于癌旁正常组织（ 21.03±5.17 vs 40.67±7.92），差异有统计学意义（ t=12.285，P<0.001），且具有低表达预后差的临床特征（ Logrank P=0.032）。过表达 miRNA-195抑制了宫颈癌细胞的增殖（ t=6.725~21.433，均 P＜ 0.01）和迁移速率（ t=12.443，16.749，均 P<0.001）。CCND2和 MYB是 miRNA-195的靶基因，过表达 miRNA-195显著抑制了 CCND2和 MYB mRNA的蛋白表达（ P<0.01）。宫颈癌组织中 CCND2较癌旁正常组织显著高表达（ 52.67±4.79 vs 39.86±6.39），差异有统计学意义（ t=12.453，P<0.001）；MYB较癌旁正常组织显著高表达（ 43.06±6.43 vs 22.07±6.85），差异有统计学意义（t=13.217，P<0.001）；且分别与 miRNA-195表达呈负相关（ r=-0.726，-0.592，均 P<0.05）。过表达 CCND2和 MYB显著促进了宫颈癌细胞的增殖和迁移速率，敲低 CCND2和 MYB表达则得到与之相反的结果（ F=144.947，875.160，均 P<0.001）；在过表达 miRNA-195细胞中分别回补过表达 CCND2和 MYB后细胞增殖、迁移速率基本回归到正常水平。结论　miRNA-195可通过靶向调控 CCND2和 MYB表达抑制宫颈癌癌细胞的增殖和迁移，进而参与宫颈癌的发生发展。
Abstract:
Objective　By detecting the expression of miRNA-195 in cervical cancer tissues, the effect and mechanism ofmiRNA-195 on the proliferation and migration of cervical cancer cells were explored. Methods　The miRNA-195 in 35 clinicalcervical cancer tissues and corresponding adjacent normal tissues were detected, and the relationship between stage and prognosisof cervical cancer patients was analyzed by Kaplan-Meier Plotter database. Cell proliferation assay and scratch migration assaywere used to verify the effects of miRNA-195 on the proliferation and migration of cervical cancer Siha and HeLa cells. Thetarget genes of miRNA-195 were predicted by microRNA database, and the targeted binding relationship was verified by doubleluciferase gene assay. The regulation of miRNA-195 on target genes was verified by qRT-PCR assay. The expression of targetgenes in cervical cancer and their correlation with miRNA-195 were analyzed. Cell complement assay was used to verify whethermiRNA-195 plays a role in cervical cancer by targeting protein expression regulation. Results　The relative expression ofmiRNA-195 in cervical cancer tissues was lower than that in adjacent normal tissues (21.03±5.17 vs 40.67±7.92), thedifference was statistically significance(t=12.285, P<0.001), and had the clinical characteristics of low expression and poorprognosis (Logrank P=0.032). Overexpression of miRNA-195 inhibited proliferation (t=6.725~21.433, all P<0.01) and migrationrate (t =12.443, 16.749, P<0.001) of cervical cancer cells. CCND2 and MYB were target genes of miRNA-195, and the mRNAand protein expressions of CCND2 and MYB were significantly inhibited by overexpression of miRNA-195 (P<0.01). CCND2was significantly higher in cervical cancer tissues than in adjacent normal tissues (52.67±4.79 vs 39.86±6.39), the differencewas statistically significance(t=12.453, P<0.001). The expression of MYB was significantly higher than that of adjacentnormal tissues (43.06±6.43 vs 22.07±6.85), the difference was statistically significance(t=13.217, P<0.001). The expression ofmiRNA-195 was negatively correlated with miRNA-195 expression (r=-0.726, -0.592, all P <0.05). Overexpression of CCND2and MYB significantly promoted the proliferation and migration rate of cervical cancer cells, while knockdown expression ofCCND2 and MYB showed opposite results (F=144.947，875.160, all P<0.001). After the overexpression of CCND2 and MYBin the overexpressing miRNA-195 cells, the cell proliferation and migration rate basically returned to the normal level.Conclusion　In cervical cancer, miRNA-195 can inhibit the proliferation and migration of cancer cells by targeting CCND2 andMYB expression, and participate in the occurrence and development of cervical cancer.

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备注/Memo

备注/Memo:
基金项目：河北省医药卫生科技发展计划项目（2019 WS 317）。
作者简介：郝艳芳（1985-），女，本科，医师，研究方向：妇科肿瘤临床和基础研究，E-mail:Iu8wye36@163.com。
更新日期/Last Update: 1900-01-01