长链非编码RNA HULC沉默表达对人脑胶质母细胞瘤细胞SHG44增殖和凋亡的影响

Effects of Silencing Expression of Long Non-Coding RNA HULC on Proliferation and Apoptosis of Human Glioblastoma Cell SHG44

【目的】探讨长链非编码RNA肝癌高表达转录本(LncRNA HULC)沉默表达对人脑胶质母细胞瘤细胞株SHG44增殖和凋亡的影响。【方法】实时荧光定量PCR(qRT-PCR)验证HULC沉默表达组(HULC-siRNA)及其阴性对照组(NC组)HULC的表达水平。CCK8增殖实验和平板克隆形成实验检测瘤细胞的增殖能力。细胞周期和细胞凋亡实验检测瘤细胞的周期分布及凋亡能力。【结果】qRT-PCR证实HULC-siRNA组较NC组的HULC表达量显著降低(P=0.003)。CCK8增殖实验显示HULC-siRNA组较NC组细胞增殖率显著降低(第2天P=0.003;第3天P=0.005;第4天P=0.009)。平板克隆形成实验显示NC组和HULC-siRNA组的克隆形成率分别为(34.11±1.24)%,(14.44±0.87)%,沉默表达HULC后细胞克隆形成率显著降低(P<0.001)。细胞周期实验显示NC组和HULC-siRNA组的细胞数分别为G1期(36.89±4.09、51.74±0.68),S期(46.95±2.49、36.89±2.13),沉默表达HULC后细胞周期明显被阻滞在G1/S期(G1期P=0.023,S期P=0.038)。细胞凋亡实验显示NC组和HULC-siRNA组的早期凋亡率分别为(2.57±0.22)%,(7.063±0.71)%,沉默表达HULC后细胞早期凋亡率显著增加(P=0.004)。【结论】LncRNA HULC沉默表达后可抑制胶质母细胞瘤细胞的增殖,促进其凋亡。

【Objective】To explore the effects of long-chain non-coding RNA highly up-reglated in liver cancer(LncRNA HULC)silencing expression on proliferation and apoptosis of human glioblastoma cell line SHG44.【Methods】Quantitative Real-time Polymerase Chain Reaction(qRT-PCR)was used to verify the expression level of HULC in HULC silent expression group(HULC-siRNA)and negative control group(NC group). CCK8 proliferation assay and plate colony formation assay were used to detect the proliferation of glioblastoma cells. Cell cycle and apoptosis assays were used to detect the cell cycle distribution and apoptosis of glioblastoma cells.【Results】qRT-PCR confirmed that HULC-siRNA group had significantly lower expression of HULC than NC group(P=0.003). CCK8 proliferation experiment showed that the proliferation rate of HULC-siRNA group was significantly lower than that of NC group on the second,third and fourth days of experiment(Day 2,P=0.003;Day 3,P=0.005;Day 4,P=0.009). Plate colony formation assay showed the cloning rates in the NC and HULC-siRNA groups were(34.11 ± 1.24)% and(14.44 ± 0.87)%,and it showed that the cell clone formation rate was clearly decreased after silenced expression of HULC(P<0.001). The cell cycle assay showed that the numbers of cells in NC group and HULC-siRNA group were G1 phase(36.89 ± 4.09,51.74± 0.68)and S phase(46.95 ± 2.49,36.89 ± 2.13),and it showed that the cell cycle was blocked in G1/S phase after silencing HULC expression apoptotic rates of NC group and HULC-siRNA group were(2.57 ± 0.22)% and(7.063 ± 0.71)%,and it showed that the early apoptotic rate of the cells was significantly increased after silencing HULC expression(P=0.004).【Conclusion】Silencing of LncRNA HULC can inhibit the proliferation of glioblastoma cells and promote their apoptosis.