摘要
目的:探讨盐酸石蒜碱(lycorine hydrochloride,LH)对肝癌HCCLM3细胞恶性生物学行为的影响及其对circASH2L/miR-124-3p轴的调控作用。方法:将HCCLM3细胞分为不同浓度LH处理组(LH-L、LH-M、LH-H组)和Con、si-NC、si-circASH2L、LH+pcDNA、LH+pcDNA-circASH2L组;以CCK-8法、平板克隆形成实验、流式细胞术、细胞划痕实验和Transwell小室实验分别检测HCCLM3细胞的增殖、克隆形成、凋亡、迁移和侵袭;qPCR法检测HCCLM3细胞中circASH2L和miR-124-3p的表达量;双荧光素酶报告基因实验检测circASH2L和miR-124-3p的靶向关系;WB法检测cleaved-caspase3、cleaved-caspase9、E-cadherin、N-cadherin蛋白表达量。结果:与Con组比较,LH不同浓度组细胞增殖抑制率升高(P<0.05),凋亡率与cleaved-caspase3、cleavedcaspase9、E-cadherin蛋白水平升高(均P<0.05),miR-124-3p的表达水平升高(P<0.05),克隆形成数与侵袭细胞数减少(均P<0.05),划痕愈合率与N-cadherin蛋白水平降低(P<0.05),circASH2L的表达水平降低(P<0.05),且不同浓度组间比较差异有统计学意义(P<0.05)。circASH2L可负向调控miR-124-3p。与si-NC组比较,si-circASH2L组细胞增殖抑制率升高(P<0.05),凋亡率与cleaved-caspase3、cleaved-caspase9、E-cadherin蛋白水平升高(均P<0.05),划痕愈合率与N-cadherin蛋白水平降低(均P<0.05),克隆形成数与侵袭细胞数减少(均P<0.05)。与LH+pcDNA组比较,LH+pcDNA-circASH2L组miR-124-3p的表达水平降低(P<0.05),细胞增殖抑制率降低(P<0.05),凋亡率与cleaved-caspase3、cleaved-caspase9、E-cadherin蛋白水平降低(均P<0.05),克隆形成数与侵袭细胞数增多(均P<0.05),划痕愈合率与N-cadherin蛋白水平升高(均P<0.05)。结论:LH可通过调控circASH2L/miR-124-3p轴来抑制肝癌细胞HCCLM3的增殖、迁移、侵袭并诱导其凋亡。
Objective:To explore the effect of lycorine hydrochloride(LH)on the malignant biological behaviors of liver cancer HCCLM3 cells and its regulation on circASH2 L/miR-124-3 p axis.Methods:HCCLM3 cells treated with different concentrations of LH were divided into LH groups with different concentrations(LH-L group,LH-M group and LH-H group),Con group,si-NC group,si-circASH2 L group,LH+pcDNA group,LH+pcDNA-circASH2 L group.CCK-8 assay,plate clone formation test,flow cytometry,wound-healing assay,and Transwell assay were used to detect the proliferation,clone formation,apoptosis,migration and invasion of HCCLM3 cells.qPCR was used to detect the expression levels of circASH2 L and miR-124-3 p in HCCLM3 cells.Dual luciferase reporter gene assay was used to experiment detect the targeting relationship between circASH2 L and miR-124-3 p.WB was used to detect the expression of cleaved-caspase3,cleaved-caspase9,E-cadherin,and N-cadherin.Results:Compared with the Con group,the cell proliferation inhibition rates of LH groups with different concentrations were increased(P<0.05),the apoptosis rates and the expression levels of cleaved-caspase3,cleaved-caspase9 and E-cadherin were increased(all P<0.05),the expression levels of miR-124-3 p were increased(P<0.05),the number of clone formation and invasive cells was decreased(all P<0.05),the wound-healing rates and the expression levels of N-cadherin were decreased(all P<0.05),and the expression levels of circASH2 L were decreased(P<0.05),and the difference among different concentration groups was statistically significant(all P<0.05).CircASH2 L could negatively regulate miR-124-3 p.Compared with the si-NC group,the cell proliferation inhibition rates of the si-circASH2 L group was increased(P<0.05),and the apoptosis rate and the expression levels of cleaved-caspase3,cleaved-caspase9 and E-cadherin were increased(all P<0.05),the wound-healing rate and the expression levels of N-cadherin were decreased(all P<0.05),and the number of clone formation and invasive cells was decreased(all P<0.05).Compared with the LH+pcDNA group,the expression level of miR-124-3 p in the LH+pcDNA-circASH2 L group was decreased(P<0.05),the cell proliferation inhibition rate was decreased(P<0.05),the apoptosis rate and the expression levels of cleaved-caspase3,cleaved-caspase9 and E-cadherin were decreased(all P<0.05),the number of clone formation and invasive cells was increased(P<0.05),and the wound-healing rate and expression level of N-cadherin were increased(all P<0.05).Conclusion:LH can inhibit the proliferation,migration and invasion of liver cancer HCCLM3 cells and induce their apoptosis by regulating the circASH2 L/miR-124-3 p axis.
作者
苏珊珊
王怀璋
唐玉君
万家鑫
SU Shanshan;WANG Huaizhang;TANG Yujun;WAN Jiaxin(Oncology Department,The Second Clinical Medical College of Henan University of Chinese Medicine,Zhengzhou 450011,Henan,China;Department of Traditional Chinese and Western Medicine,Henan Cancer Hospital,Zhengzhou 450003,Henan,China;Anorectal Department,The Second Clinical Medical College of Henan University of Chinese Medicine,Zhengzhou 450018,Henan,China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2021年第10期969-977,共9页
Chinese Journal of Cancer Biotherapy
基金
河南省科技攻关计划资助项目(No.162102310331)
河南省重点科技攻关计划资助项目(No.142102310128)。
