Objective: To explore the role of Long noncoding RNA UFC1 (lincRNA-UFC1) in modulating the metastasis and invasion of hepatocellular carcinoma (HCC) cells and the underlying mechanism.
Methods: Human HCC cell line Huh7 was infected with the lentiviral vector carrying lincRNA-UFC1 to obtain a cell line with lincRNA-UFC1 overexpression. A short hairpin RNA (shRNA) targeting lincRNA-UFC1 was delivered in human HCC BEL-7402 cells via a lentiviral vector to obtain a cell line with lincRNA-UFC1 knockdown. Expression levels of lincRNA-UFC1 in the two HCC cell lines were detected using real-time PCR, and the changes in the cell invasion and migration in response to lincRNA-UFC1 overexpression or knockdown were analyzed using Transwell and wound-healing assays. The expressions of GSK-3β/β-catenin-related proteins in the cells were detected with Western blotting. XAV-939, a GSK-3β/β-catenin inhibitor, was used for assessing the impact of lincRNAUFC1 overexpression on the invasion and migration of the HCC cells through Transwell and wound-healing assays.
Results: Overexpression of lincRNA-UFC1 significantly promoted the invasion and migration of Huh7 cells as compared with the control cells (P < 0.001), while lincRNA-UFC1 knockdown obviously suppressed the invasion and migration of BEL-7402 cells (P < 0.001). The results of Western blotting showed that the expressions of proteins associated with the cell invasion and migration, namely β-catenin and P-GSK-3β, were significantly upregulated in response to lincRNA-UFC1 overexpression, and were obviously lowered after lincRNA-UFC1 knockdown. Treatment of the cells with XAV-939 significantly reversed the effect of lincRNA-UFC1 overexpression on the cell invasion and migration (P < 0.001).
Conclusions: lincRNA-UFC1 overexpresison promotes cell invasion and migration through the GSK-3β/β-catenin axis in HCC cells in vitro.
目的: 探讨长链非编码RNA-UFC1在肝癌侵袭和转移中的作用及机制。
方法: 应用lincRNA-UFC1慢病毒载体转染人肝癌细胞株Huh7构建lincRNA-UFC1过表达组和对照组；应用lincRNA-UFC1干扰载体转染人肝癌细胞株BEL-7402构建lincRNA-UFC1干扰组和干扰对照组（scramble），通过实时定量PCR实验分析两种肝癌细胞中lincRNA-UFC1的表达水平。Transwell及划痕实验检测lincRNA-UFC1过表达组及干扰组相比对照组侵袭迁移能力的变化。Western blot检测lincRNAUFC1过表达组及干扰组相比对照组中GSK-3β/β-catenin蛋白的表达变化。利用GSK-3β/β-catenin信号通路抑制剂XAV939阻断后，观察Transwell及划痕实验中lincRNA-UFC1过表达对肝癌细胞侵袭迁移能力的影响。
结果: 在肝细胞癌中，过表达lincRNA-UFC1相较于对照组，肝癌细胞的侵袭转移能力明显增加（P＜0.001），而沉默lincRNA-UFC1的表达相较于scramble组，肝癌细胞侵袭转移能力明显减弱（P＜0.001）。Western blot结果显示，过表达lincRNA-UFC1与对照组相比较，侵袭转移相关蛋白β-catenin和p-GSK-3β表达明显上调（P＜0.001），而lincRNA-UFC1沉默表达后，结果相反。利用GSK-3β/β-catenin信号通路抑制剂XAV939处理肝癌细胞可以逆转lincRNA-UFC1促进肝癌细胞侵袭转移的功能（P＜0.001）。
结论: 在肝细胞癌中lincRNA-UFC1通过上调磷酸化GSK-3β及β-catenin促进肝癌细胞侵袭转移。
Keywords: hepatocellular carcinoma; lincRNA-UFC1; p-GSK3; tumor progression; β-catenin.