| 孙留生,韩朝阳,李爱云.RNAi抑制iASPP基因表达对大肠癌细胞凋亡的影响及机制[J].肿瘤学杂志,2019,25(4):300-304. |
| RNAi抑制iASPP基因表达对大肠癌细胞凋亡的影响及机制 |
| Effect of Inhibiting of iASPP Gene Expression by RNA Interference on Apoptosis of Colorectal Cancer Cells |
| 投稿时间：2018-03-28 |
| DOI：10.11735/j.issn.1671-170X.2019.04.B003 |
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| 中文关键词: iASPP基因 大肠肿瘤 凋亡 STAT3信号通路 RNA干扰 |
| 英文关键词:iASPP gene colorectal cancer apoptosis STAT3 signaling pathway RNA interference |
| 基金项目: |
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| 中文摘要: |
| 摘 要：［目的］ 探讨RNAi抑制iASPP基因表达对大肠癌细胞凋亡的影响。［方法］ 参照脂质体LipofectamineTM2000转染说明转染iASPP的siRNA（iASPP-siRNA组）进入人大肠癌LoVo细胞株，并设置阴性对照组（转染无义的siRNA序列）和空白对照组（仅加入脂质体），Western bloting检测转染48h的3组细胞中iASPP的蛋白表达；CCK8法于转染的24h、48h、72h检测细胞活力；流式细胞术检测转染48h的细胞凋亡率；Western blotting检测Ki-67、含半胱氨酸的天冬氨酸蛋白水解酶3（Caspase3）、STAT3和磷酸化的信号转导与转录因子3(p-STAT3)的蛋白表达。［结果］ iASPP-siRNA组（0.108±0.013）LoVo细胞株iASPP的蛋白表达显著低于空白对照组（0.389±0.036）（P<0.05）；iASPP-siRNA组细胞在24h（0.267±0.034）、48h（0.495±0.067）和72h（0.682±0.068）的活力细胞均显著低于空白对照组（0.385±0.042、0.688±0.074、0.977±0.097）（P<0.05），在48h的细胞凋亡率（11.18%±0.78%）显著高于空白对照组（2.13%±0.45%）（P<0.05）。Ki-67（0.126±0.018）和p-STAT3（0.110±0.012）的蛋白表达水平显著低于空白对照组（0.365±0.045、0.169±0.018），Caspase3（0.197±0.020）表达水平高于空白对照组（0.086±0.009）（P<0.05），3组间STAT3蛋白表达水平差异无统计学意义（P＞0.05）。［结论］通过RNAi抑制iASPP基因表达可降低大肠癌细胞增殖，诱导细胞凋亡，机制可能是下调STAT3信号，对细胞增殖的影响是下调Ki-67表达，凋亡的影响是上调Caspase3表达。 |
| 英文摘要: |
| Abstract：［Objective］ To investigate the effect of inhibiting iASPP gene expression by RNA interference on apoptosis of colorectal cancer cells. ［Methods］ Human colorectal cancer LoVo cells were transfected with iASPP siRNA (iASPP-siRNA group)，nonsense sequence siRNA (negative control group) or liposome alone(blank control group) for 24，48 and 72h，respectively. The expression of iASPP protein in three groups was detected by Western blotting；cell viability was detected by CCK8 assay，cell apoptosis was detected by flow cytometry；the expression of Ki-67，Caspase3，STAT3 and p-STAT3 proteins was detected by Western blotting. ［Results］ The expression of iASPP protein in iASPP-siRNA group(0.108±0.013) was significantly lower than that in blank control group(0.389±0.036)(P<0.05). The cell activity in iASPP-siRNA group at 24h (0.267±0.034)，48h (0.495±0.067) and 72h(0.682±0.068) were significantly decreased compared with the control group(0.385±0.042，0.688±0.074，0.977±0.097). Cell apoptosis rate(11.18%±0.78%) increased significantly at 48h compared with the blank control group(2.13%±0.45%)%. The expression of Ki-67(0.126±0.018) and p-STAT3 protein(0.110±0.012) was significantly decreased compared with the control group(0.365±0.045，0.169±0.018)，and the Caspase3 protein expression(0.197±0.020) was significantly increased compared with the control group(0.086±0.009)(all P<0.05). There was no significant difference in protein expression of STAT3 among three groups(P>0.05). ［Conclusion］ Inhibition of iASPP expression by RNA interference can reduce the proliferation of colorectal cancer cells and induce cell apoptosis，which is associated with the down-regulation of p-STAT3 and Ki-67 expression，and up-regulation of Caspase3 expression. |
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