| 陆巍,王飞,梁柳芬.黏蛋白 1基因小干扰 RNA联合丙泊酚抑制肺癌 H460细胞生长的机制研究[J].安徽医药,2021,25(3):441-445. |
| 黏蛋白 1基因小干扰 RNA联合丙泊酚抑制肺癌 H460细胞生长的机制研究 |
| Study on the mechanism of MUC1 gene siRNA combined with propofol inhibiting the growth of lung cancer H460 cells |
| |
| DOI：10.3969/j.issn.1009-6469.2021.03.004 |
| 中文关键词: 肺肿瘤 肿瘤细胞，培养的 MUC1基因 丙泊酚 肺癌细胞 生长 Akt信号通路 |
| 英文关键词: Lungneoplasms Tumorcells,cultured MUC1gene Propofol Lungcancercells Growth Aktsignalingpathway |
| 基金项目: |
|
| 摘要点击次数: 1497 |
| 全文下载次数: 459 |
| 中文摘要: |
| 目的探讨黏蛋白 1(MUC1)基因小干扰 RNA(siRNA)联合丙泊酚对肺癌 H460细胞生长的抑制及可能的机制。方法 siRNA干扰人肺癌 H460细胞株 MUC1基因，采用实时荧光定量 PCR(qPCR)和蛋白质印迹法( Western blotting)法检测转染效果；以不同浓度的丙泊酚干预 H460细胞，细胞计数试剂盒( CCK-8)检测丙泊酚对 H460细胞活力的影响，筛选丙泊酚作用浓度；实验分为转染对照( si-NC)组、转染 MUC1 siRNA(si-MUC1)组、丙泊酚干预( Propofol)组和转染 MUC1 siRNA联合丙泊酚干预(si-MUC1+Propofol)组；采用 CCK-8检测 H460细胞增殖情况，克隆形成实验分析 H460细胞克隆形成能力， Western blotting检测 H460细胞中增殖细胞核抗原(PCNA)和细胞周期蛋白 D1(cyclin D1)、蛋白激酶 B(protein kinase B，Akt)和磷酸化 Akt(p-Akt)的表达水平。结果 si-NC组和 si-MUC1组细胞中 MUC1 mRNA的表达分别为:(1.00±0.10)、(0.18±0.02)，MUC1蛋白表达分别为:(0.56±0.06)、(0.21±0.02)转染 MUC1 siRNA的 H460细胞中 MUC1 mRNA和蛋白表达水平显著降低(t=24.122，P<0.001，t= 16.602，P<0.001)；不同浓度的丙，泊酚对 H460细胞有不同程度的抑制作用，并筛选出半数致死浓度 55.96 μmol/L的丙泊酚用于后续实验； si-NC组、 si-MUC1组、 Propofol组和 si-MUC1+Propofol组细胞克隆形成数分别为:(163.36±8.22)个、(101.26±7.69)个、(92.64±6.88)个、(52.09±6.14)个， si-MUC1组和 Propofol组细胞活力、克隆形成能力、 PCNA、cyclin D1和 p-Akt的表达水平均显著低于 si-NC组( P<0.05)高于 si-MUC1+Propofol组( P<0.05)。结论敲低 MUC1基因和丙泊酚均能抑制肺癌 H460细胞生长，两者联合对 H460细胞生长，抑制作用更显著，其作用机制可能与抑制 Akt信号通路的激活有关。 |
| 英文摘要: |
| Objective To investigate the inhibitory effect of mucin 1 (MUC1) gene small interfering RNA (siRNA) combined withpropofol on the growth of lung cancer H460 cells and its possible mechanism. Methods siRNA interfered with the MUC1 gene of hu? man lung cancer H460 cell line. Real-time fluorescent quantitative PCR (qPCR) and Western blotting (Western blotting) were used todetect the transfection effect; different concentrations of propofol were used to intervene in H460 cells. Cell counting kit (CCK-8) De?tect the effect of propofol on the viability of H460 cells, and screen the concentration of propofol; the experiment was divided into trans?fection control (si-NC) group, transfection MUC1 siRNA (si-MUC1) group, propofol intervention (Propofol) ) Group and transfected MUC1 siRNA combined with propofol intervention (si-MUC1+Propofol) group; CCK-8 was used to detect the proliferation of H460cells, the clone formation experiment was used to analyze the cloning ability of H460 cells, and the proliferating cell nuclear antigen inH460 cells was detected by Western blotting (PCNA) and cyclin D1 (cyclin D1), protein kinase B (protein kinase B, Akt) and phosphor?ylated Akt (p-Akt) expression levels. Results The expression of MUC1 mRNA in the cells of the si-NC group and si-MUC1 groupwere: (1.00±0.10), (0.18±0.02), and the MUC1 protein expression were: (0.56±0.06), (0.21±0.02), transfection The expression levels ofMUC1 mRNA and protein in H460 cells of MUC1 siRNA were significantly reduced (t=24.122, P<0.001, t=16.602, P<0.001); differentconcentrations of propofol have different degrees of inhibition on H460 cells, and screening Propofol with a lethal concentration of 55.96 μmol/L was used in subsequent experiments; the number of cell clones in the si-NC group, si-MUC1 group, Propofol group and si-MUC1+Propofol group were: (163.36±8.22) Cells, (101.26±7.69) cells, (92.64±6.88) cells, (52.09±6.14) cells, cell viability, clono?genic ability, PCNA, cyclin D1 and p-Akt expression levels in si-MUC1 and Propofol groups were significant Lower than si-NC group (P <0.05), higher than si-MUC1+Propofol group (P<0.05). Conclusions Both knockdown of MUC1 gene and propofol can inhibit the growth of H460 cells in lung cancer. The combination of the two has a more significant inhibitory effect on inhibits the growth of H460cells, and its mechanism may be related to the inhibition of the activation of Akt signaling. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
