过表达lncRNA HCG18调控miR-107/CDKN2B轴对甲状腺癌细胞增殖、迁移和侵袭的机制研究

目的 探讨长链非编码RNA人类白细胞抗原复合体18(lncRNA HCG18)调控miR-107/周期蛋白依赖激酶抑制因子2B(CDKN2B)轴对甲状腺乳头状癌(PTC)细胞增殖、侵袭、迁移的影响。方法 实时荧光定量PCR(qRT-PCR)、Western blot分别检测60例PTC患者癌组织、癌旁组织及人正常甲状腺上皮细胞TEC及人PTC细胞WRO、TPC-1、SW579中HCG18、miR-107、CDKN2B蛋白表达；将WRO细胞分为CK组、si-NC组、si-HCG18组、pcDNA组、pcDNA-HCG18组、pcDNA-HCG18+mimic NC组、 pcDNA-HCG18+miR-107 mimic组，qRT-PCR检测各组细胞HCG18、miR-107表达；CCK-8法检测细胞增殖；划痕实验检测细胞迁移；Transwell检测细胞侵袭；Western blot检测CDKN2B、细胞周期素D1 (CyclinD1)、基质金属蛋白酶(MMP)-2、MMP-9蛋白表达；双荧光素酶实验验证HCG18和miR-107、miR-107和CDKN2B的关系。结果 在PTC组织和P...

lncRNA HCG18 regulates the proliferation,migration and invasion of thyroid cancer cells through the miR-107/CDKN2B axis

Objective To investigate the influences of long non-coding RNA human leukocyte antigen complex 18 (lncRNA HCG18) on the proliferation, invasion and migration of papillary thyroid carcinoma (PTC) cells by regulating miR-107/cyclin-dependent kinase inhibitor 2B (CDKN2B) axis. Methods Real-time fluorescence quantitative PCR (qRT-PCR) and Western blot were used to assess the protein expression of HCG18, miR-107 and CDKN2B in cancer tissues and adjacent tissues of 60 PTC patients, human normal thyroid epithelial cells TEC and human PTC cells WRO, TPC-1 and SW579. WRO cells were divided into CK group, si-NC group, si-HCG18 group, pcDNA group, pcDNA-HCG18 group, pcDNA-HCG18+mimic NC group and pcDNA-HCG18+miR-107 mimic group. The expressions of HCG18 and miR-107 in each group were detected by qRT-PCR. Cell proliferation was detected by CCK-8 assay. Scratch assay was used to assess cell migration. Transwell test was used to assess cell invasion. Western blot was used to assess CDKN2B, cyclin D1 (CyclinD1), matrix metalloproteinase (MMP)-2, and MMP-9 protein expression; and dual luciferase experiments was used to verify the correlations between HCG18 and miR-107, miR-107 and CDKN2B. Results In PTC tissues and PTC cells, the HCG18 and CDKN2B proteins were expressed at a low level, and miR-107 was highly expressed. In WRO cells, the protein levels of HCG18 and CDKN2B were the lowest, and the expression of miR-107 was the highest (P<0.05). Compared with the si-NC group, the protein expressions of HCG18 and CDKN2B in the si-HCG18 group were significantly decreased, and the expression of miR-107 was significantly increased (P<0.05). Compared with the pcDNA group, the protein expression of HCG18 and CDKN2B in the pcDNA-HCG18 group were significantly increased, and the expression of miR-107 was significantly reduced (P<0.05). Compared with the pcDNA-HCG18+mimic NC group, the pcDNA-HCG18+miR-107 mimic group showed no significant difference in the expression of HCG18 (P>0.05), but the expression of miR-107 was increased and the protein expression of CDKN2B were significantly reduced (P<0.05). Up-regulation of HCG18 could inhibit the proliferation, invasion, and migration of WRO cells and the protein expressions of CyclinD1, MMP-2, MMP-9; while down-regulation of HCG18 showed the opposite trend. Over-expression of miR-107 attenuated the inhibitory effects on WRO cell proliferation, invasion and migration by up-regulating HCG18. There was a targeted regulatory correlations between HCG18 and miR-107, and between miR-107 and CDKN2B. Conclusion Over-expression of HCG18 may up-regulate the expression of CDKN2B by sponging miR-107, thereby inhibiting the proliferation, migration and invasion of PTC cells.