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地锦草乙醇提取物通过circRHOT1/miR-29a-3p轴抑制结直肠癌SW480细胞的恶性生物学行为
引用本文:樊红革,戴卫波,张学峰. 地锦草乙醇提取物通过circRHOT1/miR-29a-3p轴抑制结直肠癌SW480细胞的恶性生物学行为[J]. 中国肿瘤生物治疗杂志, 2022, 29(9): 806-812
作者姓名:樊红革  戴卫波  张学峰
作者单位:中山市中医院 a. 肛肠科;b. 中药药理实验室;c. 第三外科,广东 中山 528400
基金项目:广东省中医药局中医科研项目(No. 20201371)
摘    要:目的:探讨地锦草乙醇提取物(EEEH)对人结直肠癌SW480细胞生物学行为的影响及其分子机制。方法:体外培养SW480细胞,实验分为Con组、EEEH-L组、EEEH-M组、EEEH-H组、si-NC组、si-circRHOT1组、EEEH-H+pcDNA组、EEEH-H+pcDNA-circRHOT1组,分别以si-NC、si-circRHOT1、pcDNA、pcDNA-circRHOT1转染SW480细胞,采用CCK-8法、细胞克隆形成实验、Transwell实验分别检测转染后各组细胞的增殖、迁移及侵袭能力,qPCR法检测转染后各组SW480细胞circRHOT1和miR-29a-3p的表达,WB法检测各组细胞中MMP-2、MMP-9蛋白的表达。双荧光素酶报告基因实验检测circRHOT1与miR-29a-3p之间的靶向关系。结果:与Con组比较,EEEH-L组、EEEH-M组、EEEH-H组SW480细胞中MMP-2和MMP-9蛋白表达均明显降低(均P<0.05),circRHOT1的表达均降低(均P<0.05)而miR-29a-3p的表达均升高(均P<0.05)且呈剂量依赖性;...

关 键 词:结直肠癌  SW480细胞  地锦草  乙醇提取物  circRHOT1  miR-29a-3p  增殖  迁移  侵袭
收稿时间:2022-03-27
修稿时间:2022-07-16

Ethanol extract from Euphorbia humifusa inhibits the malignant biological behaviors of colorectal cancer SW480 cells by regulating the circRHOT1/miR-29a-3p molecular axis
FAN Hongge,DAI Weibo,ZHANG Xuefeng. Ethanol extract from Euphorbia humifusa inhibits the malignant biological behaviors of colorectal cancer SW480 cells by regulating the circRHOT1/miR-29a-3p molecular axis[J]. Chinses Journal of Cancer Biotherapy, 2022, 29(9): 806-812
Authors:FAN Hongge  DAI Weibo  ZHANG Xuefeng
Affiliation:a. Department of Anorectal Medicine; b. Laboratory of Pharmacology of Traditional Chinese Medicine; c. the Third Department of Surgery, Zhongshan Hospital of Traditional Chinese Medicine, Zhongshan 528400, Guangdong, China
Abstract:Objective: To explore the effect of ethanol extract from euphorbia humifusa (EEEH) on the biological behaviors of human colorectal cancer SW480 cells and its molecular mechanism. Methods: Human colorectal cancer SW480 cells were cultured in vitro,divided into the following experimental groups: Con group, EEEH-L group, EEEH-M group, EEEH-H group,si-NC group, si-circRHOT1 group, EEEH-H+pcDNA group and EEEH-H+pcDNA-circRHOT1 group and transfected with si-NC, si-circRHOT1, pcDNA and PCDNA-circRHOT1 respectively. CCK-8 assay, cell clone formation assay, Transwell assay were used to detect cell proliferation,migration and invasion ability in each group after transfection. qPCR method was employed to detect the expression of circRHOT1 and miR-29a-3p in all the groups of SW480 cells after transfection, and the expressions of MMP-2 and MMP-9 proteins were detected by WB. The dual luciferase reporter experiment was used to detect the targeting relationship between circRHOT1 and miR-29a-3p.Results: Compared with the Con group, the protein expressions of MMP-2 and MMP-9 in SW480 cells in EEEH-L, EEEH-M, and EEEH-H groups were significantly decreased (all P<0.05), the expression of circRHOT1 decreased (all P<0.05) and the expression of miR-29a-3p increased (all P<0.05) in a dose-dependent manner. The cell survival rate, the number of cell clone formation and the number of migration and in asive cells were decreased (all P<0.05). circRHOT1 can target and negatively regulate the expression of miR-29a-3p. Knockdown of circRHOT1 can inhibit the proliferation, migration and invasion of SW480 cells, while overexpression of circRHOT1 can attenuate the inhibitory effects of EEEH on the proliferation, migration and invasion of SW480 cells.Conclusion: EEEH can inhibit the proliferation, migration and invasion of colorectal cancer SW480 cells by regulating the circRHOT1/miR-29a-3p axis.
Keywords:colorectal cancer   SW480 cell   Euphorbia humifusa   ethanol extract   circRHOT1   miR-29a-3p   proliferation   migration   invasion
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