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袁小波, 肖桂香, 唐静, 李丽娟, 李雨静.款冬花多糖通过调控miR-4282表达抑制宫颈癌细胞增殖、迁移侵袭的机制研究[J].湖南中医药大学学报,2024,44(6):991-1000[点击复制] |
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| 款冬花多糖通过调控miR-4282表达抑制宫颈癌细胞增殖、迁移侵袭的机制研究 |
| 袁小波,肖桂香,唐静,李丽娟,李雨静 |
| (湖南交通工程学院护理与医学技术学院, 湖南 衡阳 421219;郴州市第一人民医院产科, 湖南 郴州 423000) |
| 摘要: |
| 目的 探讨款冬花多糖通过调控miR-4282表达抑制宫颈癌细胞增殖、迁移侵袭的机制。方法 体外培养人宫颈癌细胞株HeLa和C-33A并以10、20、40、80、120 mg·L-1的款冬花多糖处理，然后通过CCK-8实验测定各组细胞存活率并筛选款冬花多糖最佳作用浓度。将HeLa和C-33A细胞随机分为对照组、5-氟尿嘧啶（5-fluorouracil, 5-FU）组、款冬花多糖组、miR-4282 mimics组、阴性对照组、款冬花多糖+miR-4282 inhibitor组，分组处理后通过RT-PCR检测各组细胞miR-4282表达；通过CCK-8实验、EDU实验、流式细胞实验分别检测各组细胞增殖、凋亡；通过划痕实验、Transwell实验检测各组细胞迁移、侵袭情况；通过Western blot法检测各组细胞增殖相关蛋白（Cyclin D1）、凋亡相关蛋白[B淋巴细胞瘤-2（B-cell lymphoma-2, Bcl-2）、Bcl-2关联X蛋白（B-cell lymphoma-2 related X protein, Bax）]、上皮-间质转化相关蛋白[Vimentin、基质金属蛋白酶2（matrix metallo-proteinases 2, MMP-2）、E-cadherin]表达。结果 与对照组相比，5-FU组、款冬花多糖组、miR-4282 mimics组细胞存活率、EDU阳性率、迁移率、迁移数、侵袭数、Cyclin D1与Bcl-2、Vimentin、MMP-2蛋白表达降低（P<0.05），miR-4282表达、凋亡率、Bax与E-cadherin蛋白表达升高（P<0.05）；阴性对照组细胞各指标无明显变化（P＞0.05）。与款冬花多糖组相比，款冬花多糖+miR-4282 inhibitor组细胞存活率、EDU阳性率、迁移率、迁移数、侵袭数、Cyclin D1与Bcl-2、Vimentin、MMP-2蛋白表达升高（P<0.05），miR-4282表达、凋亡率、Bax与E-cadherin蛋白表达降低（P<0.05）。结论 款冬花多糖可通过上调miR-4282表达而抑制宫颈癌细胞增殖、迁移侵袭并促使其凋亡，最终对其起到明显抑癌作用。 |
| 关键词: 款冬花多糖 miR-4282表达 宫颈癌 增殖 迁移 侵袭 |
| DOI：10.3969/j.issn.1674-070X.2024.06.009 |
| 投稿时间：2023-12-27 |
| 基金项目:衡阳市科学技术局2022年度指导性计划项目（202222035745）。 |
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| Mechanism of polysaccharide of Farfarae Flos in inhibiting proliferation, migration, and invasion of cervical cancer cells by regulating miR-4282 expression |
| YUAN Xiaobo, XIAO Guixiang, TANG Jing, LI Lijuan, LI Yujing |
| (School of Nursing and Medical Technology, Hunan Institute of Traffic Engineering, Hengyang, Hunan 421219, China;Obstetrics Department of the First People's Hospital of Chenzhou, Chenzhou, Hunan 423000, China) |
| Abstract: |
| Objective To explore the mechanism of polysaccharide of Farfarae Flos in inhibiting the proliferation, migration, and invasion of cervical cancer cells by regulating miR-4282 expression. Methods Human cervical cancer cell lines HeLa and C-33A were cultured in vitro and treated with 10, 20, 40, 80, and 120 mg·L-1 of polysaccharide of Farfarae Flos. CCK-8 assay was used to determine the survival rate of each group of cells and screen the optimal concentration of polysaccharide of Farfarae Flos. HeLa and C-33A cells were randomly grouped into control group, fluorouracil (5-FU) group, polysaccharide of Farfarae Flos group, miR-4282 mimics group, negative control group, and polysaccharide of Farfarae Flos+miR-4282 inhibitor group. The miR-4282 expression was examined by RT-PCR; the cell proliferation and apoptosis were deterrmined by CCK-8 assay, EDU assay, and flow cytometry; the cell migration and invasion were checked by scratch and transwell assays; immunoblotting was used to examine the expression of cell proliferation related proteins (Cyclin D1), apoptosis related proteins (Bax, Bcl-2), epithelial mesenchymal transition-related proteins [Vimentin, Matrix Metallo-proteinases 2 (MMP2), E-cadherin]. Results Compared with the control group, the cell survival rate, EDU positivity rate, migration rate, migration number, invasion number, the Cyclin D1 protein expression as well as protein expressions of Bcl-2, Vimentin, and MMP-2 in 5-FU group, polysaccharide of Farfarae Flos group, and miR-4282 mimics group decreased (P<0.05), while the miR-4282 expression, apoptosis rate, and protein expressions of Bax and E-cadherin increased (P<0.05). There was no significant change in various indicators of cells in the negative control group (P>0.05). Compared with the polysaccharide of Farfarae Flos group, the cell survival rate, EDU positivity rate, migration rate, migration number, invasion number, the Cyclin D1 protein expression as well as protein expressions of Bcl-2, Vimentin, and MMP-2 in the polysaccharide of Farfarae Flos+miR-4282 inhibitor group were higher (P<0.05), while the miR-4282 expression, apoptosis rate, and protein expressions of Bax and E-cadherin were lower (P<0.05). Conclusion The polysaccharide of Farfarae Flos can inhibit the proliferation, migration, invasion, and apoptosis of cervical cancer cells by up-regulating the miR-4282 expression, ultimately playing a significant role in cancer suppression. |
| Key words: polysaccharide of Farfarae Flos miR-4282 expression cervical cancer proliferation migration invasion |
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