卵巢癌组织中miR-223的表达及其对卵巢癌OVCAR3细胞增殖和侵袭的促进作用

doi:10.13481/j.1671-587X.20230119

摘要/Abstract

摘要：

目的分析微小RNA-223（miR-223）在卵巢癌组织中的表达情况，探讨miR-223对卵巢癌OVCAR3细胞增殖和侵袭的影响，并阐明其分子调控机制。方法采用实时荧光定量PCR（RT-qPCR）法检测45例卵巢癌患者手术切除组织样本和不同卵巢癌细胞中miR-223表达水平，分析不同临床病理特征卵巢癌患者癌组织中miR-223表达水平。体外培养卵巢癌OVCAR3细胞，采用生物信息学、双荧光素酶报告基因实验和Western blotting法分析miR-223对N-myc下游调节基因1（NDRG1）的靶向调控关系。OVCAR3细胞分别转染阴性对照模拟物（NC组）、miR-223 抑制剂（MiR in组）和同时转染miR-223 抑制剂及siRNA-NDRG1质粒（MiR in+si-NDRG1组），克隆形成实验检测各组细胞克隆形成数，流式细胞术检测各组细胞凋亡率，Transwell小室实验检测各组细胞中侵袭细胞数。结果与癌旁正常组织比较，卵巢癌组织中miR-223表达水平明显升高（P<0.01），miR-223表达水平与卵巢癌患者组织学分化程度、FIGO分期和淋巴结转移有密切关联（P<0.01）。不同卵巢癌细胞中miR-223表达水平均高于正常卵巢上皮细胞（P<0.01）。miR-223可靶向结合NDRG1并抑制NDRG1的表达，且在卵巢癌组织中NDRG1 mRNA表达水平与miR-223表达水平呈负相关关系（r=－0.291，P<0.01）。与NC组比较，MiR in组细胞中克隆形成数和侵袭细胞数均明显减少（P<0.05或P<0.01），细胞凋亡率明显升高（P<0.01）；与MiR in组比较，MiR in+si-NDRG1组细胞中克隆形成数和侵袭细胞数明显增加（P<0.05或P<0.01），细胞凋亡率明显降低（P<0.01）。结论 MiR-223在卵巢癌组织中呈高表达，其表达水平与卵巢癌的严重程度有密切关联。miR-223可通过靶向抑制NDRG1表达促进卵巢癌细胞增殖和侵袭，并抑制细胞凋亡。

关键词: miR-223, N-myc下游调节基因1, 卵巢肿瘤, 细胞增殖, 细胞侵袭

Abstract:

Objective To analyze the expression of microRNA-223 （miR-223） in the ovarian cancer tissue and explore the effect of miR-223 on the proliferation and invasion of ovarian cancer OVCAR3 cells， and to clarify the molecular regulatory mechanism. Methods The expression levels of miR-223 in tissue samples taken during surgical removal of 45 patients with ovarian cancer and different ovarian cancer cells were detected by real-time fluorescence quantitative PCR （RT-qPCR） method， and the expression levels of miR-223 in cancer tissue of the ovarian cancer patients with different clinicopathological features were analyzed. The ovarian cancer OVCAR3 cells were cultured in vitro； bioinformatics， dual luciferase reporter gene experiment and Western blotting method were used to analyze the targeted regulation relationship of miR-223 to N-myc downstream regulatory gene 1 （NDRG1）. The OVCAR3 cells were transfected with negative control mimic（NC group），miR-223 inhibitor（MiR in group） and miR-223 inhibitor and siRNA-NDRG1 plasmid at the same time（MiR in+si-NDRG1 group）. The number of clone formation was detected by colony formation test， the apoptotic rate was measured by flow cytometry， and the number of invasion cells was measured by Transwell chamber assay. Results Compared with adjacent normal tissue the expression level of miR-223 in cancer tissue was significantly increased（P<0.01）；the miR-223 expression level was closely related to the degree of histological differentiation， FIGO stage and lymph node metastasis of ovarian cancer patients （P<0.01）. Meanwhile， the expression levels of miR-223 in the different ovarian cancer cells were higher than that in normal ovarian epithelial cells（P<0.01）. MiR-223 could target NDRG1 and inhibit the expression of NDRG1， and the expression level of NDRG1 mRNA in ovarian cancer tissue was negatively correlated with the expression level of miR-223 （r=－0.291， P<0.01）. Compared with NC group， the number of clone formation and the number of invasion cells in MiR in group were decreased （P<0.05 or P<0.01）， while the apoptotic rate was significantly increased（P<0.01）. Compared with MiR in group， the number of clone formation and the number of invasion cells in MiR in + si-NDRG1 group were increased （P<0.05 or P<0.01）， while the apoptotic rate was decreased （P<0.01）. Conclusion MiR-223 is highly expressed in the ovarian cancer tissue， which is positively correlated with the severity of ovarian cancer. MiR-223 can promote the proliferation and invasion of ovarian cancer cells and inhibit apoptosis by targeted inhibition of NDRG1 expression.

Key words: MiR-223, N-myc downstream regulatory gene 1, Ovarine neoplasms, Cell proliferation, Cell invasion

中图分类号:

R737.31

陈艳雅,招锦兰,李婵,黄丽珊. 卵巢癌组织中miR-223的表达及其对卵巢癌OVCAR3细胞增殖和侵袭的促进作用[J]. 吉林大学学报(医学版), 2023, 49(1): 150-157.

Yanya CHEN,Jinlan ZHAO,Chan LI,Lishan HUANG. Expression of miR-223 in ovarian cancer tissue and its promoting effect on proliferation and invasion of ovarian cancer OVCAR3 cells[J]. Journal of Jilin University(Medicine Edition), 2023, 49(1): 150-157.

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参考文献 24

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Clinicopathological feature	n	MiR-223	t	P
Age（year）			0.982	0.331
<50	18	4.02±2.26
≥50	27	4.82±2.90
Tumor size（d/cm）			0.599	0.552
<5	21	4.25±2.42
≥5	24	4.73±2.90
Histological differentiation			2.812	0.007
Medium/high	28	3.69±2.35
Low	17	5.84±2.69
FIGO stage			5.355	<0.001
Ⅰ-Ⅱ	22	2.80±1.57
Ⅲ-Ⅳ	23	6.13±2.49
Lymph node metastasis			3.505	0.001
No	26	3.44±2.39
Yes	19	5.96±2.37

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