医学分子生物学杂志 ›› 2025, Vol. 22 ›› Issue (5): 435-441.doi: 10.3870/j.issn.1672-8009.2025.05.004

• 论著 • 上一篇    下一篇

过表达 miR-193b-5p 介导自噬逆转宫颈癌细胞顺铂化疗耐药的作用研究 #br#

  

  1. 新疆维吾尔自治区人民医院1科研教育中心,3妇科诊疗中心 乌鲁木齐市, 830001 2新疆医科大学第一附属医院药学部 乌鲁木齐市, 830011
  • 出版日期:2025-09-30 发布日期:2025-10-09
  • 基金资助:
    新疆维吾尔自治区卫生健康委 天山英才医药卫生高层次人才项目 (No. TSYC202401B147)

Effect of Overexpression of miR-193b-5p on Autophagy to Reverse Cisplatin Chemotherapy Resistance in Cervical Cancer Cells #br#

  1. 1Research and Education Center,3 Gynecological Diagnosis and Treatment Center, Xinjiang Uygur Autonomous Region Peoples Hospital, Urumqi, 830001, China 2 Department of Pharmacy, the First Affiliated Hospital of Xinjiang Medical University, Urumqi,830011, China
  • Online:2025-09-30 Published:2025-10-09

摘要: 目的 探究过表达微小 RNA-193b-5p (miR-193b-5p) 对人宫颈癌顺铂 ( cisplatin, DDP) 耐药细胞株 SiHa / DDP 化疗耐药性的影响及机制。 方法 培养人宫颈癌细胞株 SiHa 和人宫颈癌顺铂耐药细胞株SiHa / DDP, MTT 法检测不同浓度 (5、 10、 15、 20、 25、 30 μmol / L) DDP 处理后两种细胞的存活率, 测定半抑制浓度 (IC50 ) 值。 将实验分为 SiHa 组、 SiHa / DDP 组、 mimics NC 组 (转染 mimics NC 的 SiHa / DDP细胞)、 miR-193b-5p mimics 组 (转染 miR-193b-5p mimics 的 SiHa / DDP 细胞), 实时荧光定量 PCR ( RTqPCR) 法检测细胞中 miR-193b-5p 表达。 DDP 处理上述细胞后, MTT 法和流式细胞术检测各组细胞存活率、 凋亡率, 细胞免疫荧光染色检测细胞中 LC3B 表达, 蛋白质印迹实验检测各组细胞中 LC3Ⅱ / LC3Ⅰ 、Beclin-1、 P62 蛋白质表达。 结果 与 SiHa 细胞比较, 同一浓度 DDP 处理下的 SiHa / DDP 细胞存活率显著升高 (P< 0. 05), SiHa / DDP 细胞的 IC50值显著升高 (P< 0. 05), 而细胞中 miR-193b-5p 相对表达量显著下调 (P< 0. 05)。 细胞转染后, 与对照组和 mimics NC 组比较, miR-193b-5p mimics 组 SiHa / DDP 细胞中 miR-193b-5p 相对表达量显著上调 (P< 0. 05)。 与 SiHa / DDP 组、 mimics NC 组比较, miR-193b-5p mimics 组 SiHa / DDP 细胞存活率显著下降、 细胞凋亡率显著增加 ( P< 0. 05), 细胞中 LC3B 荧光强度明显减弱, LC3Ⅱ / LC3Ⅰ 比值、 Beclin-1 蛋白质相对表达量显著下调 ( P< 0. 05), P62 蛋白质相对表达量显著上调 ( P<0. 05)。 结论 miR-193b-5p 在人宫颈癌顺铂耐药细胞株 SiHa / DDP 中表达下降, 其过表达能够逆转 SiHa /DDP 细胞对 DDP 的耐药性, 该作用可能与其抑制细胞自噬水平有关。

关键词:

宫颈癌, 微小 RNA-193b-5p, 顺铂, 化疗耐药, 自噬

Abstract: Objective To investigate the effect and mechanism of overexpression of microRNA-193b-5p ( miR-193b-5p) on cisplatin ( DDP) -resistant cell line SiHa / DDP in human cervicalcancer. Methods Human cervical cancer cell line ( SiHa) and its cisplatin-resistant cell line( SiHa / DDP) were cultured. The survival rates of the two cell lines treated with DDP (5, 10, 15, 20, 25, 30 μmol / L) were detected by MTT assay, and the half inhibitory concentration ( IC50 ) value was determined. Cells were divided into 4 groups: SiHa group, SiHa / DDP group, mimics NC group ( SiHa / DDP cells transfected with mimics NC), miR-193b-5p mimics group ( SiHa / DDP cells transfected with miR-193b-5p mimics). The expression level of miR-193b-5p was detected by RT-qPCR. Cells were then treated with DDP, and the cell survival rate and apoptosis rate were detected by the MTT method and flow cytometry. The expression of microtubule-associated protein 1 light chain 3 B (LC3B) was detected by cell immunofluorescence staining, and the protein expression levels of LC3Ⅱ / LC3Ⅰ , Beclin-1, and P62 were detected by Western blotting. Results Thesurvival rate of SiHa / DDP cells was higher than that of the SiHa cells when treated with DDP at thesame concentration (P< 0. 05), and the IC50 value of cells was significantly increased (P< 0. 05).The relative expression level of miR-193b-5p in the SiHa / DDP cells was significantly downregulated(P< 0. 05). The relative expression level of miR-193b-5p in the miR-193b-5p mimics group was higher than that in the control group and mimics NC group (P< 0. 05). Compared with those in theSiHa / DDP group and the mimics NC group, the survival rate of SiHa / DDP cells in the miR-193b-5p mimics group was significantly decreased, the apoptosis rate was significantly increased ( P <0. 05), the fluorescence intensity of LC3B in cells was significantly weakened, the LC3Ⅱ / LC3Ⅰratio and the relative expression level of Beclin-1 protein were significantly downregulated ( P <0. 05), and the relative expression level of P62 protein was significantly upregulated (P < 0. 05).Conclusion The expression level of miR-193b-5p was decreased in the cisplatin-resistant cell lineSiHa / DDP of human cervical cancer. Its overexpression could reverse the resistance of SiHa / DDPcells to DDP, which may be related to its inhibition of cell autophagy.

Key words:

cervical cancer, microRNA-193b-5p, cisplatin, chemotherapy resistance, autophagy

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