国际肿瘤学杂志

国际肿瘤学杂志 ›› 2025, Vol. 52 ›› Issue (11): 673-679.doi: 10.3760/cma.j.cn371439-20241203-00116

• 论著 • 上一篇    下一篇

circRNA-15430靶向miR-10对胃癌细胞增殖及侵袭的影响

章帅, 刘靓靓, 黄迪, 盛茹, 齐梦瑶, 李曙光()   

  1. 河北北方学院附属第一医院胃肠肿瘤外科，张家口 075000
  • 收稿日期:2024-12-03 修回日期:2025-05-06 出版日期:2025-11-08 发布日期:2025-12-21
  • 通讯作者: 李曙光 E-mail:lishuguang196508@163.com
  • 基金资助:
    张家口市科学技术局科技计划(2322042D)

Effects of circRNA-15430 targeting miR-10 on proliferation and invasion of gastric cancer cells

Zhang Shuai, Liu Liangliang, Huang Di, Sheng Ru, Qi Mengyao, Li Shuguang()   

  1. Department of Gastrointestinal Oncological Surgery， First Affiliated Hospital of Hebei North University， Zhangjiakou 075000， China
  • Received:2024-12-03 Revised:2025-05-06 Online:2025-11-08 Published:2025-12-21
  • Contact: Li Shuguang E-mail:lishuguang196508@163.com
  • Supported by:
    Zhangjiakou Science and Technology Bureau Science and Technology Plan(2322042D)

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摘要：

目的 探究环状RNA（circRNA）-15430通过调控微RNA-10（miR-10）对胃癌细胞增殖及侵袭的影响。方法 实时荧光定量PCR检测胃上皮细胞株GES-1及4种胃癌细胞株（GTL16、NUGC-3、MGC803、Snu-1）中circRNA-15430、miR-10的相对表达水平。取胃癌细胞株GTL16，将其分为胃癌细胞（GC）组、胃癌细胞+circRNA-15430-NC（CN）组、胃癌细胞+circRNA-15430抑制剂（CI）组、胃癌细胞+ circRNA-15430激动剂（CM）组、胃癌细胞+miR-10-NC（MN）组、胃癌细胞+miR-10激动剂（MM）组、胃癌细胞+miR-10抑制剂（MI）组、胃癌细胞+circRNA-15430激动剂+miR-10抑制剂（UI）组，CCK-8法检测细胞增殖，Transwell小室检测细胞侵袭，双荧光素酶报告基因实验证实circRNA-15430和miR-10的相互作用。结果 实时荧光定量PCR结果显示，circRNA-15430在GES-1和4种胃癌细胞中的相对表达量分别为1.00±0.00、0.31±0.02、0.62±0.04、0.59±0.03、0.63±0.05，差异有统计学意义（F=334.70，P<0.001）；miR-10 在GES-1和4种胃癌细胞中的相对表达量分别为1.00±0.00、1.80±0.18、1.53±0.14、1.49±0.13、1.51±0.15，差异有统计学意义（F=27.52，P<0.001）。circRNA-15430在4种胃癌细胞中的相对表达量均低于GES-1细胞，在NUGC-3、MGC803、Snu-1细胞中的相对表达量均高于GTL16细胞（均P<0.05），miR-10在4种胃癌细胞中的相对表达量均高于GES-1细胞（均P<0.05），在NUGC-3、MGC803、Snu-1细胞中的相对表达量均低于GTL16细胞（均P<0.05），因此选取GTL16细胞进行后续实验。细胞增殖和侵袭实验结果显示，GC组、CN组、CI组、CM组的细胞增殖率分别为（47.06±4.61）%、（47.04±4.63）%、（68.61±6.59）%、（28.52±2.16）%，差异有统计学意义（F=71.02，P<0.001）；侵袭数量分别为（195.57±20.04）、（195.35±20.08）、（316.86±32.20）、（169.04±16.11）个，差异有统计学意义（F=50.18，P<0.001）。CI组细胞增殖率、侵袭数量均高于GC组、CN组，CM组细胞增殖率、侵袭数量均低于GC组、CN组、CI组（均P<0.05）。MN组、MM组、MI组细胞增殖率分别为（47.18±4.62）%、（68.68±6.36）%、（28.55±2.12）%，细胞侵袭数量分别为（195.78±20.07）、（316.90±32.21）、（169.10±16.15）个，GC组、MN组、MM组、MI组间细胞增殖率、侵袭数量差异均有统计学意义（F=73.79，P<0.001；F=50.08，P<0.001）。MM组细胞增殖率、侵袭数量均高于GC组、MN组，MI组细胞增殖率、侵袭数量均低于GC组、MN组、MM组（均P<0.05）。UI组细胞增殖率为（20.31±1.11）%，细胞侵袭数量为（107.51±10.02）个，CM组、MI组、UI组细胞增殖率、侵袭数量差异均具有统计学意义（F=39.06，P<0.001；F=36.63，P<0.001）；UI组细胞增殖率、侵袭数量均低于CM组、MI组（均P<0.05）。双荧光素酶报告基因实验结果显示，转染circRNA-15430后可显著降低miR-10-3′-UTR-WT的荧光素酶活性（t=21.22，P<0.001），但对突变基因无显著影响（t=0.85，P=0.413）。结论 circRNA-15430可有效抑制胃癌细胞增殖及侵袭，其作用机制可能与靶向抑制miR-10有关。

关键词: 胃肿瘤, 细胞增殖, 肿瘤浸润, circRNA-15430, miR-10

Abstract:

Objective To explore the effect of circular RNA （circRNA）-15430 on the proliferation and invasion of gastric cancer cells by regulating mircoRNA-10 （miR-10）. Methods The relative expression levels of circRNA-15430 and miR-10 in gastric epithelial cell line GES-1 and four gastric cancer cell lines （GTL16， NUGC-3， MGC803， Snu-1） were detected by real-time fluorescence quantitative PCR. The gastric cancer cell line GTL16 were taken， and the cells were divided into the gastric cancer cell （GC） group， the gastric cancer cell+circRNA-15430-NC （CN） group， the gastric cancer cell+circRNA-15430 inhibitor （CI） group， the gastric cancer cell +circRNA-15430 agonist （CM） group， the gastric cancer cell+miR-10-NC （MN） group， the gastric cancer cell+miR-10 agonist （MM） group， the gastric cancer cell+miR-10 inhibitor （MI） group， and the gastric cancer cell+circRNA-15430 agonist+miR-10 inhibitor （UI） group. Cell proliferation was detected by the CCK-8 method， and cell invasion was detected by the Transwell chamber method. The dual-luciferase reporter gene assay was used to confirm the interaction between circRNA-15430 and miR-10. Results Real-time fluorescence quantitative PCR showed that the relative expression levels of circRNA-15430 in GES-1 and four types of gastric cancer cells were 1.00±0.00， 0.31±0.02， 0.62±0.04， 0.59±0.03 and 0.63±0.05， respectively， with a statistically significant difference （F=334.70， P<0.001）； The relative expression levels of miR-10 in GES-1 and four types of gastric cancer cells were 1.00±0.00， 1.80±0.18， 1.53±0.14， 1.49±0.13 and 1.51±0.15， respectively， with a statistically significant difference （F=27.52， P<0.001）. The relative expression levels of circRNA-15430 in the four types of gastric cancer cells were all lower than those in GES-1 cells， and the relative expression levels in NUGC-3， MGC803， and Snu-1 cells were all higher than those in GTL16 cells （all P<0.05）. The relative expression levels of miR-10 in the four types of gastric cancer cells were all higher than those in GES-1 cells （all P<0.05）， and the relative expression levels in NUGC-3， MGC803， and Snu-1 cells were all lower than those in GTL16 cells （all P<0.05）. Therefore， GTL16 cells were selected as the experimental cells for this experiment. The results of the cell proliferation and invasion assay showed that the cell proliferation rates of the GC group， CN group， CI group and CM group were （47.06±4.61） %， （47.04±4.63） %， （68.61±6.59） % and （28.52±2.16） %， respectively， with a statistically significant difference （F=71.02， P<0.001）. The invasion numbers were 195.57±20.04， 195.35±20.08， 316.86±32.20 and 169.04±16.11， respectively， with a statistically significant difference （F=50.18， P<0.001）. The cell proliferation rate and invasion number in the CI group were both higher than those in the GC group and CN group， while the cell proliferation rate and invasion number in the CM group were both lower than those in the GC group， CN group and the CI group （all P<0.05）. The cell proliferation rates of the MN group， MM group， and MI group were （47.18±4.62） %， （68.68±6.36） % and （28.55±2.12） %， respectively. The invasion numbers were 195.78±20.07， 316.90±32.21 and 169.10±16.15， respectively. There were statistically significant differences in cell proliferation rates and the invasion numbers among the GC group， MN group， MM group and MI group （F=73.79， P<0.001； F=50.08， P<0.001）. The cell proliferation rate and invasion number in the MM group were both higher than those in the GC group and MN group， while the cell proliferation rate and invasion number in the MI group were both lower than those in the GC group， MN group and the MM group （all P<0.05）. The cell proliferation rate in the UI group was （20.31±1.11） %， and the invasion number was 107.51±10.02. There were statistically significant differences in cell proliferation rates and invasion numbers among the CM group， MI group and UI group （F=39.06， P<0.001； F=36.63， P<0.001）. The cell proliferation rate and invasion number in the UI group were both lower than those in the CM group and MI group （all P<0.05）. The results of the dual-luciferase reporter gene assay showed that transfection with circRNA-15430 could significantly reduce the luciferase activity of miR-10-3'-UTR-WT （t=21.22， P<0.001）， but had no significant effect on the mutant gene （t=0.85， P=0.413）. Conclusions circRNA-15430 can effectively inhibit the proliferation and invasion of gastric cancer cells， and its mechanism may be related to the targeted inhibition of miR-10.

Key words: Stomach neoplasms, Cell proliferation, Neoplasm invasiveness, circRNA-15430, miR-10