| 摘要: |
| 目的探讨LncRNANNT-AS1对胃癌细胞增殖、凋亡的作用机制。方法采用实时荧光定量PCR法检测胃癌细胞系KATO-III、NUGC-3、AGS、N87及正常胃成纤维细胞系NSFC中LncRNANNT-AS1的表达水平；将KATO-III细胞系分成对照组、sh-vector组及sh-NNT-AS1组，分别不感染慢病毒、感染阴性shRNA慢病毒及感染lenti-sh-NNT-AS1慢病毒。采用MTT实验检测细胞增殖情况，流式细胞术检测细胞凋亡情况，Westernblot检测细胞凋亡蛋白Caspase3、8、9的表达水平，并进行组间比较。结果胃癌细胞系KATO-III、NUGC-3、AGS、N87LncRNANNT-AS1的表达水平均明显高于正常胃成纤维细胞系NSFC（均P＜0.01）。细胞培养12、24、48、72h后，相比sh-vector组、对照组，sh-NNT-AS1组吸光度值（OD490值）明显降低（均P＜0.05）。细胞培养7d后，sh-NNT-AS1组克隆形成数明显低于sh-vector组、对照组（均P＜0.05）。sh-NNT-AS1组细胞凋亡率明显高于sh-vector组、对照组（均P＜0.01）。sh-NNT-AS1组细胞凋亡蛋白Caspase3、9表达水平均高于sh-vector组（均P＜0.05），而Caspase8表达水平比较差异无统计学意义（P＞0.05）。结论LncRNANNT-AS1促进胃癌细胞增殖，抑制凋亡，机制可能与上调凋亡蛋白Caspase3、9表达有关。 |
| 关键词: LncRNA NNT-AS1 胃癌 增殖 凋亡 |
| DOI：10.12056/j.issn.1006-2785.2018.40.5.2017-2255 |
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| Effect of LncRNA NNT-AS1 on proliferation and apoptosis of gastric carcinoma cells and its mechanism |
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the Affiliated Hospital,College of Medicine, Ningbo University
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| Abstract: |
| Objective To investigate the effect of LncRNA NNT-AS1 on proliferation and apoptosis of gastric carcinoma cells and its mechanism. Methods The expression levels of LncRNA NNT-AS1 in the gastric carcinoma cell lines KATO-III, NUGC-3, AGS, N87 and normal gastric cell line NSFC was determined by qRt-PCR. The KATO-III cells were transfected with negative shRNA-Lenti (vector group) or Lenti-sh-NNT-AS1 (sh-NNT-AS1 group). The cell proliferation ability was measured by MTT assay, apoptosis was tested by flow cytometry, the expression of Caspase 3, 8, 9 protein was measured by Western blot. Results The relative express levels of LncRNA NNT-AS1 in KATO-III, NUGC-3, AGS and N87 cells were significantly higher
than that in normal gastric NSFC cells (P＜0.05). MTT assay demonstrated that the OD490 value of sh-NNT-AS1 group was
significantly lower than control and vector group after culturing for 12, 24, 48 and 72 h (P＜0.05). The number of clone formation cells in sh-NNT-AS1 group was significantly lower than that in control group and vector group (P＜0.01). The apoptosis rate of sh-NNT-AS1 group was significantly higher than that in vector and control group (P＜0.01). The expression level of cleaved Caspase 3, 9 protein in sh-NNT-AS1 group was significantly higher than that in vector group (P＜0.05, P＜0.01); while, there was no significantly difference in expression of cleaved caspase-8 protein between sh-NNT-AS1 and vector groups (P ＞0.05). Conclusion LncRNA NNT-AS1 may promote proliferation and inhibit apoptosis of gastric carcinoma cells, which may is
associated with up-regulation of Caspase 3 and 9. |
| Key words: LncRNA NNT-AS1 Gastric carcinoma Proliferation Apoptosis |
