常春藤皂苷元通过调控lncRNA PCAT19/miR-4319对TPC-1甲状腺癌细胞生物学行为的影响

Effects of hederagenin on the biological behavior of TPC-1 thyroid cancer cells via PCAT19/miR-4319 regulation

目的 探讨常春藤皂苷元对甲状腺癌细胞生物学行为的影响及其可能作用机制。方法 用不同剂量常春藤皂苷元处理人甲状腺癌TPC-1细胞,si-NC、si-PCAT19分别转染至TPC-1细胞,pcDNA、pcDNA-PCAT19分别转染至TPC-1细胞后加入常春藤皂苷元处理细胞。MTT、流式细胞术分别检测细胞增殖和周期,Transwell检测细胞迁移及侵袭,RT-qPCR法检测PCAT19、miR-4319表达,双荧光素酶报告实验检测PCAT19、miR-4319靶向关系,Western blot法检测Twist、Snail、N-cadherin、E-cadherin蛋白表达。结果 常春藤皂苷元能剂量依赖性地降低细胞存活率、S期细胞比例、PCAT19 mRNA表达及Twist、Snail、N-cadherin蛋白表达(P<0.05),减少迁移及侵袭细胞数(P<0.05),升高G_(0)/G_(1)期细胞比例、E-cadherin蛋白表达、miR-4319表达(P<0.05)。PCAT19可靶向调控miR-4319的表达。转染si-PCAT19可抑制TPC-1细胞增殖、迁移及侵袭,并可诱导细胞周期阻滞于G_(0)/G_(1)期。转染pcDNA-PCAT19可通过抑制miR-4319表达而恢复常春藤皂苷元对TPC-1细胞生物学行为的作用。结论 常春藤皂苷元可通过调节PCAT19/miR-4319表达而诱导细胞周期阻滞于G_(0)/G_(1)期,并抑制甲状腺癌细胞增殖、迁移及侵袭。

AIM To explore the effects of hederagenin on the biological behavior of thyroid cancer cells and its possible mechanism.METHODS The human thyroid carcinoma TPC-1 cells were treated with different doses of hederagenin;or transfected with si-NC and si-PCAT19,respectively;or treated with hederagenin following the pcDNA and pcDNA-PCAT19 transfection,respectively.TPC-1 cells had their cell proliferation and cell cycle detected by MTT and flow cytometry;their cell migration and invasion detected by Transwell;their PCAT19 and miR-4319 expressions detected by RT-qPCR;their targeting relationship of PCAT19 and miR-4319 detected by luciferase reporter assay;and their protein expressions of Twist,Snail,N-cadherin and E-cadherin detected by Western blot method.RESULTS Hederagenin dose-dependently reduced the cellular viability,S-phase cell ratio,the expression of PCAT19 and the protein expressions of Twist,Snail and N-cadherin(P<0.05);reduced the number of migrating and invasive cells(P<0.05),and increased the ratio of cells in G_(0)/G_(1) phase,the expression of E-cadherin protein and miR-4319(P<0.05).PCAT19 targeted specifically at miR-4319 expression.Transfection of si-PCAT19 inhibited the proliferation,migration and invasion of TPC-1 cells,and induced cell cycle arrest in G_(0)/G_(1) phase.The pcDNA-PCAT19 transfection restored the effect of hederagenin on the biological behavior of TPC-1 cells by inhibiting the expression of miR-4319.CONCLUSION With its effects in induction of cell cycle arrest in G_(0)/G_(1) phase via regulation on PCAT19/miR-4319 expression,hederagenin inhibits the proliferation,migration and invasion of thyroid cancer cells.