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目的 分析熊果酸对甲状腺乳头状癌(Papillary thyroid carcinoma, PTC)长链非编码RNA(Long chain non coding RNA,lncRNA)-LINC01089及其靶基因表达的影响。方法 LINC01089为前期通过转录组测序筛选并通过验证的差异表达lncRNA,体外常规培养甲状腺乳头状癌TPC-1细胞、正常人甲状腺细胞(Nthy-Ori3-1)。将细胞分为正常组(正常人甲状腺细胞组)、对照组(TPC-1细胞组)、不同浓度熊果酸(Ursolic acid, UA)干预组(10、20、40μmol·L-1)。CCK-8法检测各组甲状腺乳头状癌TPC-1细胞的存活率；划痕实验、Transwell实验检测不同浓度UA对TPC-1细胞水平、垂直迁移以及侵袭的影响作用；利用RT-qPCR法检测TPC-1细胞中LINC01089的相对表达量及熊果酸的干预作用；RT-qPCR及Western blot检测靶基因丝状伪足相关基因(Ras homolog family member F,RHOF)、4-羟基苯丙酮酸双加氧酶(4-Hydroxyphenylpyruvate dioxygenase, HPD)的mRNA及蛋白表达水平以及熊果酸的干预作用。结果 不同浓度的UA(10、20、40、80μmol·L-1)作用不同时间(12～48 h)后均能抑制TPC-1细胞的增殖，当UA浓度为10～40μmol·L-1时，在12～24 h内随着浓度的增加和作用时间的延长，细胞的抑制效应逐渐增强，差异具有统计学意义(P<0.05),UA处理TPC-1细胞24 h后，细胞增殖缓慢，随着UA(10、20、40μmol·L-1)浓度的增高，细胞出现凋亡的形态学改变；与对照组相比，UA(10、20、40μmol·L-1)组可抑制TPC-1细胞迁移和侵袭，呈明显浓度依赖性，差异具有统计学意义(P<0.01)。熊果酸能不同程度上调LINC01089的表达量，下调RHOF、HPD的mRNA及蛋白表达水平。结论 熊果酸对甲状腺乳头状癌细胞具有一定的抑制作用，其机制可能通过上调LINC01089作用于靶基因RHOF发挥抑癌效应。
Abstract：Objective To analyze the effect of ursolic acid on the expressions of long chain non coding RNA(lncRNA)-LINC01089 and its target genes in papillary thyroid carcinoma(PTC). Method LINC01089 was a differentially expressed lncRNA that was previously screened and verified by transcriptome sequencing in the early stage. TPC-1 cells and normal human thyroid cells(Nthy-ori3-1) were routinely cultured in vitro and were divided into normal human thyroid cell group, control group and UA groups(10,20,40 μmol·L-1). The survival rate of TPC-1 cells in each group was detected by CCK-8 assays. Scratch and Transwell experiments were conducted to investigate the effects of different concentrations of ursolic acid on the horizontal, vertical migration and invasion of TPC-1 cells. RT qPCR method was used to detect the relative expression level of LINC01089 in TPC-1 cells and the intervention effect of ursolic acid. RT-qPCR and Western blot were used to detect the mRNA and protein expression levels of target genes Ras Homolog Family Member F(RHOF) and 4-Hydroxyphenylpyruvate Dioxygenase(HPD) as well as the intervention effect of ursolic acid. Results Different concentrations of UA(10,20,40 and 80 μmol·L-1) could inhibit the proliferation of TPC-1 cells for different periods of time(12～48 hours). When the concentration of ursolic acid was 10～40 μmol·L-1,the inhibitory effect of the cells gradually increased with increasing of concentration and prolongation of action time within 12 and 24 hours(P<0.05). After treating TPC-1 cells with ursolic acid for 24 hours, the cell proliferation was slow, and as the concentration of ursolic acid(10,20 and 40 μmol·L-1) increased, morphological changed in cell apoptosis. Compared with the control group, the ursolic acid(10,20,40 μmol·L-1) groups inhibited the migration and invasion of TPC-1 cells in a significant concentration dependent manner(P<0.01). Ursolic acid can up-regulate the expression of LINC01089,and down-regulate the mRNA and protein expression levels of RHOF and HPD. Conclusion Ursolic acid has a certain inhibitory effect on thyroid papillary carcinoma cells, and its mechanism may be related with the LINC01089 and its target gene RHOF.
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基本信息:
DOI：10.13193/j.issn.1673-7717.2025.09.003
中图分类号:R285
引用信息:
[1]伊琳,何亚丽,吕红英等.熊果酸对甲状腺乳头状癌细胞LINC01089及其靶基因表达的影响[J].中华中医药学刊,2025,43(09):11-15+259-262.DOI:10.13193/j.issn.1673-7717.2025.09.003.
基金信息:
国家自然科学基金项目(82160842); 甘肃省自然科学基金项目(20JR10RA368,24JRRA590); 甘肃省人民医院院内科研资金项目(23GSSYD-17)