circRNA CCND1对人肝癌HepG2细胞增殖、凋亡、侵袭和迁移的影响

Effects of circRNA CCND1 on proliferation, apoptosis, invasion and migration of human hepatocellular carcinoma HepG2 cells

摘要:

目的 探讨circRNA CCND1对人肝癌HepG2细胞增殖、凋亡、侵袭和迁移的影响。方法 采用荧光原位杂交（FISH）实验及实时荧光定量聚合酶链反应（qRT-PCR）检测circRNA CCND1在HepG2细胞及L02细胞中的表达情况；进一步干扰及过表达circRNA CCND1后，通过qRT-PCR检测circRNA CCND1在HepG2细胞中的表达情况，CCK-8法和EDU法检测HepG2细胞增殖情况，流式细胞术检测细胞凋亡、细胞周期，Transwell实验检测细胞侵袭情况，划痕实验检测细胞迁移情况。结果 HepG2细胞circRNA CCND1相对表达量较LO2细胞高（P <0.05）。Si-circRNA CCND1组circRNA CCND1相对表达量较Control组和Si-NC组下降（P <0.05），Oe-circRNA CCND1组较Control组和Oe-NC组升高（P <0.05）。各组细胞培养0、24、48、72和96 h后细胞增殖活性比较，经重复测量设计的方差分析，结果 ①不同时间点细胞增殖活性比较，差异有统计学意义（P <0.05）；②各组细胞增殖活性比较，差异有统计学意义（P <0.05）；③各组细胞增殖活性变化趋势比较，差异有统计学意义（P <0.05）。与Control组及Si-NC组比较，Si-circRNA CCND1组细胞凋亡率升高（P <0.05），G₀/G₁期比值升高（P <0.05），S及G₂/M期比值下降（P <0.05）；与Control组及Oe-NC组比较，Oe-circRNA CCND1组细胞凋亡率下降（P <0.05），G₀/G₁期比值下降（P <0.05），S及G₂/M期比值升高（P <0.05）。Si-circRNA CCND1组细胞侵袭数、细胞迁移数较Control组、Si-NC组少（P <0.05），Oe-circRNA CCND1组较Control组及Oe-NC组多（P <0.05）。结论 circRNA CCND1能够促进人肝癌细胞HepG2细胞增殖、侵袭和迁移，抑制细胞凋亡。

Abstract:

Objective To investigate the effects of circRNA CCND1 on the proliferation, apoptosis, invasion and migration of human hepatocellular carcinoma HepG2 cells.Methods The expression of circRNA CCND1 in HepG2 and LO2 cells was detected by fluorescence in situ hybridization (FISH) and quantitative real-time polymerase chain reaction (qRT-PCR). After further interference of the expression of circRNA CCND1 or overexpression of circRNA CCND1, the level of circRNA CCND1 in HepG2 cells was detected by qRT-PCR, the proliferation of HepG2 cells was detected by the CCK-8 assay and the EdU assay, cell apoptosis and cell cycle were detected by flow cytometry, cell invasion was detected by the transwell invasion assay, and cell migration was detected by the scratch assay.Results The relative expression of circRNA CCND1 in HepG2 cells was higher than that in LO2 cells (P < 0.05). The relative expression of circRNA CCND1 in the si-circRNA CCND1 group was lower than that in the control group and the si-NC group (P < 0.05), while that in the Oe-circRNA CCND1 group was higher compared with that in the control group and the Oe-NC group (P < 0.05). The proliferation activity of cells in all groups after 0, 24, 48, 72 and 96 h of culture was compared via repeated measures analysis of variance, and the results demonstrated that the cell proliferation activity was different among the time points (P < 0.05) and the groups (P < 0.05), and that the change trend of the cell proliferation activity was also different among the groups (P < 0.05). Compared with the control group and the si-NC group, the apoptosis rate was higher (P < 0.05), the percentage of cells in the G₀/G₁ phase was higher (P < 0.05), and the percentage of cells in the S and G₂/M phase was lower in the si-circRNA CCND1 group (P < 0.05). Compared with the control group and the Oe-NC group, the apoptosis rate was lower (P < 0.05), the percentage of cells in the G0/G1 phase was lower (P < 0.05), and the percentage of cells in the S and G₂/M phase was higher in the Oe-circRNA CCND1 group (P < 0.05). The number of invaded and migrated cells in the si-circRNA CCND1 group was lower than that in the control group and the si-NC group (P < 0.05), while that in the Oe-circRNA CCND1 group was higher compared with the control group and the Oe-NC group (P < 0.05).Conclusion The circRNA CCND1 promotes the proliferation, invasion and migration, but inhibits the apoptosis of human hepatocellular carcinoma HepG2 cells.