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| 右美托咪定对人结肠癌细胞增殖和自噬的影响 |
| Effects of dexmedetomidine on proliferation and autophagy of human colon cancer cells |
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| DOI：10.12089/jca.2024.07.012 |
| 中文关键词: 结肠癌 自噬 右美托咪定 微管相关蛋白1轻链3 |
| 英文关键词: Colon cancer Autophagy Dexmedetomidine Microtubule-associated protein 1 light chain 3 |
| 基金项目:新疆维吾尔自治区自然科学基金（2021D01C207） |
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| 中文摘要: |
目的： 探讨右美托咪定对人结肠癌细胞增殖和自噬的影响。
方法： 实验一中，选择处于对数生长期的人结肠癌细胞LoVo和HCT116，将细胞分为八组：LoVo-1组（L0-1组）、LoVo+右美托咪定1 nmol/L-1组（L1-1组）、LoVo+右美托咪定10 nmol/L-1组（L10-1组）、LoVo+右美托咪定100 nmol/L-1组（L100-1组）、HCT116-1组（H0-1组）、HCT116+右美托咪定1 nmol/L-1组（H1-1组）、HCT116+右美托咪定10 nmol/L-1组（H10-1组）和HCT116+右美托咪定100 nmol/L-1组（H100-1组）。细胞加药处理24、48 h时采用CCK-8法检测细胞增殖率，细胞加药处理24 h时收集细胞，采用Western blot法检测微管相关蛋白1轻链3（LC3）-Ⅱ、自噬相关蛋白Beclin-1含量。实验二中，选择处于对数生长期的人结肠癌细胞LoVo和HCT116，将细胞分为四组：LoVo-2组（L0-2组）、LoVo+右美托咪定10 nmol/L-2组（L10-2组）、HCT116-2组（H0-2组）和HCT116+右美托咪定10 nmol/L-2组（H10-2组）。细胞加药处理24 h时，收集细胞，采用免疫荧光法观察LC3蛋白表达情况并计算LC3位点阳性率；细胞加药处理24 h时，收集细胞，采用透射电镜观察自噬体。
结果： 实验一中，与L0-1组和L1-1组比较，L10-1组和L100-1组细胞加药处理后24、48 h细胞增殖率明显降低，细胞加药处理后24 h LC3-Ⅱ、Beclin-1蛋白含量明显升高（P<0.05）。与H0-1组和H1-1组比较，H10-1组和H100-1组细胞加药处理后24、48 h细胞增殖率明显降低，细胞加药处理后24 h LC3-Ⅱ、Beclin-1蛋白含量明显升高（P<0.05）。实验二中，与L0-2组比较，细胞加药处理后24 h L10-2组LC3位点阳性率明显升高（P<0.05）。与H0-2组比较，细胞加药处理后24 h H10-2组LC3位点阳性率明显升高（P<0.05）。L0-2组和H0-2组细胞膜完整，细胞核清晰。L10-2和H10-2组细胞膜破坏，细胞器排列紊乱，可见大量自噬小体及自噬溶酶体。
结论： 右美托咪定可能通过诱导自噬，抑制结肠癌细胞的增殖。 |
| 英文摘要: |
Objective: To investigate the effects of dexmedetomidine on the proliferation and autophagy of human colon cancer cells.
Methods: In experiment 1, human colon cancer cells LoVo and HCT116 were selected and divided into eight groups: LoVo-1 group (group L0-1), LoVo + dexmedetomidine 1 nmol/L-1 group (group L1-1), LoVo + dexmedetomidine 10 nmol/L-1 group (group L10-1), LoVo + dexmedetomidine 100 nmol/L-1 group (group L100-1), HCT116-1 group (group H0-1), HCT116 + dexmedetomidine 1 nmol/L-1 group (group H1-1), HCT116 + dexmedetomidine 10 nmol/L-1 group (group H10-1) and HCT116 + dexmedetomidine 100 nmol/L-1 group (group H100-1). The cell proliferation rate was detected by CCK-8 method 24 and 48 hours after drug treatment. The cells were collected 24 hours after drug treatment, and the contents of microtubule-associated protein 1 light chain 3 (LC3)-Ⅱ and autophagy associated protein Beclin-1 were detected by Western blot method. In experiment 2, LoVo and HCT116 were selected and divided into four groups: LoVo-2 group (group L0-2), LoVo + dexmedetomidine 10 nmol/L-2 group (group L10-2), HCT116-2 group (group H0-2) and HCT116 + dexmedetomidine 10 nmol/L-2 group (group H10-2). Cells were collected 24 hours after drug treatment, LC3 protein expression was observed by immunofluorescence method, and the positive rate of LC3 site was calculated. The autophagosomes were collected and observed by transmission electron microscopy 24 hours after drug treatment.
Results: In experiment 1, the cell proliferation rate in groups L10-1 and L100-1 was significantly lower than that in groups L0-1 and L1-1 24 and 48 hours after drug treatment, the protein content of LC3-Ⅱ and Beclin-1 was significantly higher than that in groups L0-1 and L1-1 24 hours after drug treatment (P < 0.05). The cell proliferation rate in groups H10-1 and H100-1 was significantly lower than that in groups H0-1 and H1-1 24 and 48 hours after drug treatment, and the protein content of LC3-Ⅱ and Beclin-1 was significantly higher than that in groups H0-1 and H1-1 24 hours after drug treatment (P < 0.05). In experiment 2, compared with group L0-2, the positive rate of LC3 site in group L10-2 was significantly increased 24 hours after drug treatment (P < 0.05). Compared with group H0-2, the positive rate of LC3 site in group H10-2 was significantly increased 24 hours after drug treatment (P < 0.05). Groups L0-2 and H0-2 have intact cell membranes and clear nuclei. The cell membrane in groups L10-2 and H10-2 was damaged, the arrangement of organelles was disordered, and a large number of autophagosomes and autophagosomes were visible.
Conclusion: Dexmedetomidine may inhibit the proliferation of colon cancer cells by inducing autophagy. |
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