| 吕燕,王武艳,季晓黎.敲低ACTL8基因对子宫内膜癌细胞增殖与迁移的影响.[J].中南医学科学杂志.,2021,(1):20-24. |
| 敲低ACTL8基因对子宫内膜癌细胞增殖与迁移的影响 |
| Effect of knock down ACTL8 gene expression on the proliferation and migration of endometrial cancer cells |
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| DOI：10.15972/j.cnki.43-1509/r.2021.01.004 |
| 中文关键词: 子宫内膜癌 Ishikawa细胞株 肌动蛋白样蛋白8基因 侵袭 迁移 增殖 |
| 英文关键词:endometrial cancer Ishikawa cell line actin-like protein 8 gene invasion migration proliferation |
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| 摘要点击次数: 770 |
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| 中文摘要: |
| 目的探讨采用携带shRNA特异序列的载体转染子宫内膜癌细胞敲低肌动蛋白样蛋白8(ACTL8)基因对细胞增殖、侵袭迁移能力的影响及其机制。方法选取人子宫内膜癌Ishikawa细胞株进行实验研究,以携带shRNA特异序列的载体及空载质粒分别转染Ishikawa细胞株,形成敲低ACTL8基因的shACTL8组和以空载质粒转染的对照组。荧光定量聚合酶链反应(PCR)检测两组细胞中ACTL8 mRNA表达,Western blot法检测两组细胞ACTL8蛋白表达,Transwell迁移和侵袭实验分析Ishikawa细胞侵袭和迁移能力,CCK8细胞增殖实验及平板克隆实验检测细胞的增殖情况,Western blot法检测两组细胞的E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、基质金属蛋白酶-9(MMP-9)、P21蛋白的表达情况。结果shACTL8组的ACTL8 mRNA、ACTL8蛋白相对表达强度均低于对照组(P＜0.05)；shACTL8组的Ishikawa细胞侵袭、迁移水平均低于对照组(P＜0.05)；在细胞培养后1天、2天、3天,shACTL8组的Ishikawa细胞数目测定OD值显著低于对照组(P＜0.05)；平板克隆实验培养3天,可见shACTL8组培养皿中细胞生长数目明显少于对照组(P＜0.05)；shACTL8组E-cadherin蛋白、P21蛋白表达强度高于对照组(P＜0.05)；shACTL8组N-cadherin蛋白、MMP-9蛋白表达强度低于对照组(P＜0.05)。结论敲低ACTL8基因抑制子宫内膜癌Ishikawa细胞增殖、迁移、侵袭。 |
| 英文摘要: |
| To investigate the effect and mechanism of the knockdown of actin-like protein 8 (ACTL8) gene on the proliferation and migration of endometrial cancer cells. MethodsThe human endometrial cancer Ishikawa cell line was selected for experimental study. Ishikawa cell lines were transfected with vectors carrying shRNA specific sequences and empty plasmids to form shACTL8 group knocked out of ACTL8 gene and a control group transfected with empty plasmids. Fluorescence quantitative PCR was used to detect ACTL8 mRNA expression in two groups of cells. Western blot method was used to detect ACTL8 protein expression in two groups of cells. Transwell migration and invasion experiments were used to analyze the invasion and migration ability of Ishikawa cell line after culture, and CCK8 cell proliferation experiment and plate cloning experiments were used to detect the proliferation of cells. Western blot method was used to detect E-cadherin, N-cadherin, matrix metalloproteinase-9 (MMP- 9) and the expression of P21 protein. ResultsThe relative expression intensity of ACTL8 mRNA and ACTL8 protein in shACTL8 group was lower than that in control group (P＜0.05); the invasion and migration level of Ishikawa cells in shACTL8 group was lower than that in control group (P＜0.05); 1d, 2d, and 3d after cell culture, the OD value of Ishikawa cells in the control group was significantly higher than that of the shACTL8 group (P＜0.05); the plate clone experiment was cultured for 3d, which can be seen in the culture dish of the shACTL8 group, and the number of cell growth was significantly less than the control group (P＜0.05); the expression intensity of E-cadherin protein and P21 protein in shACTL8 group was higher than that in control group (P＜0.05); the expression intensity of N-cadherin protein and MMP-9 protein in shACTL8 group was lower than the control group (P＜0.05). ConclusionThe knockdown ACTL8 gene inhibits the proliferation, migration and invasion of human endometrial cancer Ishikawa cells. |
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