miR-125b通过靶向BCL-2逆转宫颈癌SiHa细胞顺铂耐药的机制

doi:10.12280/gjfckx.20260116

摘要/Abstract

摘要：

目的：探讨miR-125b是否通过靶向调控B细胞淋巴瘤-2（B-cell lymphoma-2，BCL-2）的表达来实现对宫颈癌SiHa细胞顺铂（cisplatin，DDP）耐药性的影响及其作用机制。方法：以人宫颈癌细胞（SiHa）及其DDP耐药细胞株SiHa/DDP为研究对象，设置SiHa组（SiHa细胞转染mimic-NC）、SiHa/DDP组（SiHa/DDP细胞转染mimic-NC）、SiHa+mimic-miR-125b组（SiHa细胞转染miR-125b模拟物）和SiHa/DDP+mimic-miR-125b组（SiHa/DDP细胞转染miR-125b模拟物）。采用实时荧光定量聚合酶链反应（real time fluorescent quantitative polymerase chain reaction，RT-qPCR）检测miR-125b表达，CCK-8法检测细胞活力并计算DDP的半数抑制浓度（IC₅₀），流式细胞术检测细胞凋亡，蛋白质印迹法（Western blotting）检测BCL-2蛋白的表达情况。通过双荧光素酶报告基因实验验证miR-125b与BCL-2的靶向关系。结果：与亲本SiHa组比较，耐药株SiHa/DDP组miR-125b表达及细胞凋亡率均降低，BCL-2蛋白表达升高（均P<0.05）；转染mimic-miR-125b后，与耐药株SiHa/DDP组比较，SiHa/DDP+mimic-miR-125b组miR-125b表达及细胞凋亡率均升高，BCL-2蛋白表达降低（均P<0.05）；但SiHa/DDP+mimic-miR-125b组miR-125b表达及细胞凋亡率仍低于SiHa+mimic-miR-125b组，BCL-2蛋白表达仍高于SiHa+mimic-miR-125b组（均P<0.05）。SiHa组和SiHa/DDP组对DDP的IC₅₀分别为7.21 μg/mL和34.75 μg/mL；SiHa+mimic-miR-125b和SiHa/DDP+mimic-miR-125b组的IC₅₀分别为5.13 μg/mL和12.26 μg/mL。双荧光素酶报告实验显示，miR-125b模拟物能特异性降低野生型BCL-2报告基因的荧光素酶活性（P<0.05），而对突变型无影响（P>0.05）。结论：miR-125b可通过靶向抑制BCL-2表达，促进宫颈癌SiHa细胞凋亡，从而逆转其DDP耐药性。

关键词: 宫颈肿瘤, 癌, 微RNAs, 顺铂, 抗药性，肿瘤, 细胞增殖, 细胞凋亡, 微RNA-125b, B细胞淋巴瘤-2

Abstract:

Objective: To investigate whether miR-125b affects cisplatin (DDP) resistance in cervical cancer SiHa cells by targeting the regulating the expression of B-cell lymphoma-2 (BCL-2) and to explore its underlying mechanism. Methods: Using human cervical cancer cells (SiHa) and their DDP-resistant cell line SiHa/DDP as subjects, the following groups were established: SiHa group (SiHa cells transfected with mimic-NC), SiHa/DDP group (SiHa/DDP cells transfected with mimic-NC), SiHa+mimic-miR-125b group (SiHa cells transfected with miR-125b mimic), and SiHa/DDP+mimic-miR-125b group (SiHa/DDP cells transfected with miR-125b mimic). The expression of miR-125b was detected by real time fluorescent quantitative polymerase chain reaction (RT-qPCR). Cell viability was assessed using the CCK-8 assay, and the half maximal inhibitory concentration (IC₅₀) of DDP was calculated. Apoptosis was measured by flow cytometry, and the expression of BCL-2 protein was evaluated by Western blotting. The targeting relationship between miR-125b and BCL-2 was validated using a dual-luciferase reporter assay. Results: Compared with the parental SiHa group, the resistant SiHa/DDP group exhibited reduced miR-125b expression and apoptosis rate, along with increased BCL-2 protein expression (all P<0.05). After transfection with mimic-miR-125b, compared with the resistant SiHa/DDP group, the SiHa/DDP+mimic-miR-125b group showed increased miR-125b expression and apoptosis rate, and decreased BCL-2 protein expression (all P<0.05). However, miR-125b expression and apoptosis rate in the SiHa/DDP+mimic-miR-125b group remained lower than those in the SiHa+mimic-miR-125b group, while BCL-2 protein expression remained higher (all P<0.05). The IC₅₀ values of DDP were 7.21 μg/mL and 34.75 μg/mL for the SiHa group and SiHa/DDP group, respectively. The IC₅₀ values for the SiHa+mimic-miR-125b and SiHa/DDP+mimic-miR-125b groups were 5.13 μg/mL and 12.26 μg/mL, respectively. The dual-luciferase reporter assay showed that miR-125b mimic specifically reduced the luciferase activity of the wild-type BCL-2 reporter gene (P<0.05), while it had no effect on the mutant type (P>0.05). Conclusions: miR-125b can reverse DDP resistance in cervical cancer SiHa cells by targeting and inhibiting BCL-2 expression, thereby promoting apoptosis.

Key words: Uterine cervical neoplasms, Carcinoma, MicroRNAs, Cisplatin, Drug resistance, neoplasm, Cell proliferation, Apoptosis, MicroRNA-125b, B-cell lymphoma-2

王书君, 经莉. miR-125b通过靶向BCL-2逆转宫颈癌SiHa细胞顺铂耐药的机制[J]. 国际妇产科学杂志, 2026, 53(4): 390-396.

WANG Shu-jun, JING Li. Mechanism of miR-125b Reversing Cisplatin Resistance in Cervical Cancer SiHa Cells by Targeting BCL-2[J]. Journal of International Obstetrics and Gynecology, 2026, 53(4): 390-396.

图/表 10

参考文献 22

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组别	n	miR-125b相对表达量
SiHa组	3	1.00±0.10
SiHa/DDP组	3	0.43±0.04^a
SiHa+mimic-miR-125b组	3	1.23±0.13^ab
SiHa/DDP+mimic-miR-125b组	3	0.65±0.06^bc
F		457.511
P		<0.001

组别	n	miR-125b相对表达量
SiHa组	3	1.00±0.10
SiHa/DDP组	3	0.43±0.04^a
SiHa+mimic-miR-125b组	3	1.23±0.13^ab
SiHa/DDP+mimic-miR-125b组	3	0.65±0.06^bc
F		457.511
P		<0.001

组别	n	DDP（μg/mL）						F	P	IC₅₀
组别	n	0	2.5	5	10	20	40	F	P	IC₅₀
SiHa组	3	99.91±1.09	88.50±3.22*	55.35±1.84*	37.22±2.52*	18.60±1.12*	8.81±4.21*	48.271	<0.001	7.21
SiHa+mimic-miR-125b组	3	101.13±1.03	79.80±2.15^a*	46.74±1.75^a*	29.51±2.31^a*	11.14±0.96^a*	3.35±3.87^a*	41.362	<0.001	5.13
SiHa/DDP组	3	99.94±1.16	91.06±3.28^a*	84.00±2.76^a*	75.16±2.59^a*	60.78±2.61^a*	46.37±3.48^a*	21.291	<0.001	34.75
SiHa/DDP+mimic-miR-125b组	3	99.87±0.95	88.70±3.20^b*	73.51±2.59^b*	54.38±2.67^b*	32.73±2.03^b*	24.95±1.59^b*	61.758	<0.001	12.26
F		1.001	152.294	165.580	193.061	438.746	265.557
P		0.441	<0.001	<0.001	<0.001	<0.001	<0.001

组别	n	DDP（μg/mL）						F	P	IC₅₀
组别	n	0	2.5	5	10	20	40	F	P	IC₅₀
SiHa组	3	99.91±1.09	88.50±3.22*	55.35±1.84*	37.22±2.52*	18.60±1.12*	8.81±4.21*	48.271	<0.001	7.21
SiHa+mimic-miR-125b组	3	101.13±1.03	79.80±2.15^a*	46.74±1.75^a*	29.51±2.31^a*	11.14±0.96^a*	3.35±3.87^a*	41.362	<0.001	5.13
SiHa/DDP组	3	99.94±1.16	91.06±3.28^a*	84.00±2.76^a*	75.16±2.59^a*	60.78±2.61^a*	46.37±3.48^a*	21.291	<0.001	34.75
SiHa/DDP+mimic-miR-125b组	3	99.87±0.95	88.70±3.20^b*	73.51±2.59^b*	54.38±2.67^b*	32.73±2.03^b*	24.95±1.59^b*	61.758	<0.001	12.26
F		1.001	152.294	165.580	193.061	438.746	265.557
P		0.441	<0.001	<0.001	<0.001	<0.001	<0.001

组别	n	细胞凋亡率
SiHa组	3	9.04±1.03
SiHa/DDP组	3	4.71±0.89^a
SiHa+mimic-miR-125b组	3	20.33±2.07^a
SiHa/DDP+mimic-miR-125b组	3	16.32±1.79^bc
F		63.591
P		<0.001