LINC00243靶向miR-1976促进甲状腺癌细胞增殖、迁移和侵袭

LINC00243 promotes the proliferation,migration and invasion of thyroid cancer cells by targeting miR-1976

目的:探讨长链非编码RNA(lncRNA)LINC00243对甲状腺癌细胞增殖、迁移及侵袭的影响和分子机制。方法:实时荧光定量PCR(qRT-PCR)检测正常甲状腺细胞系(HT-ori3)和甲状腺癌细胞系(BCPAP、TPC-1和SW1736)中LINC00243和miR-1976的表达水平。将LINC00243小干扰RNA(si-LINC00243)、miR-1976模拟物(miR-1976 mimics)分别转染TPC-1细胞。细胞计数试剂盒(CCK-8)检测细胞活力;Transwell实验检测细胞迁移和侵袭数量;蛋白质印记(Western blot)检测细胞周期素D1(CyclinD1)、基质金属蛋白酶2(MMP2)和MMP9的表达水平。双荧光素酶报告基因实验和qRT-PCR验证LINC00243和miR-1976的靶向调控关系。结果:与HT-ori3细胞比较,3种甲状腺癌细胞中LINC00243的表达水平显著升高,miR-1976的表达水平显著降低。沉默LINC00243或高表达miR-1976均可抑制TPC-1细胞的增殖、迁移和侵袭,抑制CyclinD1、MMP2和MMP9蛋白的表达(P<0.05)。LINC00243靶向负性调控miR-1976表达。低表达miR-1976可逆转沉默LINC00243对TPC-1细胞增殖、迁移和侵袭的抑制作用(P<0.05)。结论:在甲状腺癌细胞中,LINC00243呈高表达,miR-1976呈低表达。LINC00243通过靶向调控miR-1976促进甲状腺癌细胞增殖、迁移和侵袭。

Objective:To investigate the effects and molecular mechanisms of long-chain non-coding RNA(lncRNA)LINC00243 on proliferation,migration and invasion of thyroid cancer cells.Methods:Real-time quantitative PCR(qRT-PCR)was used to detect the expression levels of LINC00243 and miR-1976 in normal thyroid cell line(HT-ori3)and thyroid cancer cell lines(BCPAP,TPC-1 and SW1736).LINC00243 small interfering RNA(si-LINC00243)and miR-1976 mimics were transfected into TPC-1 cells,respectively.Cell viability kit(CCK-8)was used to detect cell viability.Transwell assay was used to detect cell migration and invasion.Western blot was used to detect the expression levels of CyclinD1,MMP2 and MMP9 proteins.The dual luciferase reporter gene assay and qRT-PCR were used to verify the targeted and regulatory relationship between LINC00243 and miR-1976.Results:Compared with HT-ori3 cells,the expression level of LINC00243 in the three thyroid cancer cells was significantly increased,while the expression level of miR-1976 was significantly decreased.Silencing LINC00243 or high expression of miR-1976 inhibited the proliferation,migration and invasion of TPC-1 cells,and inhibited the expression of CyclinD1,MMP2 and MMP9 proteins(P<0.05).LINC00243 targeted and negatively regulated the expression of miR-1976.Low expression of miR-1976 reversed the inhibitory effect of silencing LINC00243 on proliferation,migration and invasion of TPC-1 cells(P<0.05).Conclusion:In thyroid cancer cells,LINC00243 was highly expressed and miR-1976 was lowly expressed.LINC00243 promotes the proliferation,migration and invasion of thyroid cancer cells by targeting miR-1976.