| 摘要: |
| 目的探讨沉默Wnt/茁-catenin信号通路中的关键蛋白茁-catenin对人恶性黑色素瘤A375细胞增殖与凋亡的影响。方法将干扰慢病毒液（茁-catenin-RNAi）和空载体慢病毒液（茁-catenin-negative）分别感染A375细胞，得到的稳定感染细胞株分别标记为A375-RNAi、A375-negative。采用Westernblot法检测感染后的各组细胞中茁-catenin蛋白表达水平，MTT法检测各组细胞增殖活性，流式细胞术检测各组细胞凋亡率。结果β-catenin在A375细胞中高表达，在A375-RNAi细胞中的表达水平下降约55.1%（P＜0.05），即茁-catenin被干扰慢病毒液有效沉默；A375-negative细胞茁-catenin蛋白表达水平与A375细胞比较无统计学差异（P＞0.05）。A375-RNAi细胞与A375-negative细胞、A375细胞相比增殖活性均减弱（均P＜0.05）；A375-negative细胞增殖活性与A375细胞比较无统计学差异（P＞0.05）。A375-RNAi细胞与A375-negative细胞、A375细胞相比凋亡率明显增高（均P＜0.05）；A375-negative细胞凋亡率与A375细胞比较无统计学差异（P＞0.05）。结论慢病毒载体介导siRNA能有效沉默人黑色素细胞瘤A375细胞内茁-catenin蛋白的表达。下调茁-catenin表达能抑制A375细胞增殖，细胞凋亡率增高。 |
| 关键词: 恶性黑色素瘤 茁-catenin siRNA 增殖 凋亡 |
| DOI：10.12056/j.issn.1006-2785.2018.40.9.2018-100 |
| 分类号: |
| 基金项目:国家自然科学基金面上项目（81774313） |
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| Effects of silencing 茁-catenin expression on proliferation and apoptosis of melanoma A375 cells |
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Zhejiang Hospital of Traditional Chinese Medicine
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| Abstract: |
| Objective To investigate the effect of silencing 茁-catenin expression on the proliferation and apoptosis of malignant melanoma A375 cells. Methods The 茁-catenin-RNAi lentivirus and 茁-catenin-negative lentivirus were constructed and transfected into A375 cells(A375-RNAi and A375-negative). The expression of 茁-catenin protein was detected with Western blot; the cell proliferation was determined with MTT method; the cell apoptosis rate was examined with flow cytometry. Results Western blot showed that the protein expression of 茁-catenin in A375-RNAi cells was lower than that in A375-negative and A375
cells (P＜0.05), while the expression levels in the latter two had no significant difference (P ＞0.05). MTT showed that compared with A375 cells the proliferative activity of A375-RNAi cells was reduced (P ＜0.05), and there was no significant difference between A375-negative cells and A375 cells (P ＞0.05). Flow cytometry showed that the apoptosis rate of A375-RNAi cells was significantly higher than that in A375 cells (P＜0.05), and there was no significant difference between A375-negative cells and A375 cells (P ＞0.05). Conclusion Transfection with 茁-catenin-RNAi-LV can successfully down-regulate the expression of 茁-catenin, which leads to the inhibition of proliferation and enhancement of apoptosis in A375 cells. |
| Key words: Malignant melanoma 茁-catenin SiRNA Roliferation Poptosis |
