| 陈敏远.大黄素通过调控miRNA抑制胰腺癌新生血管形成及其机制[J].浙江中西医结合杂志,2019,29(9): |
| 大黄素通过调控miRNA抑制胰腺癌新生血管形成及其机制 |
| Emodin repressing neovascularization of pancreatic cancer by regulating microRNAs and its mechanism |
| 投稿时间：2019-04-15 修订日期：2019-08-07 |
| DOI： |
| 中文关键词: 大黄素 胰腺癌 新生血管 血管内皮生长因子 微小RNA |
| 英文关键词:Emodin, pancreatic cancer, neovascularization, VEGF, microRNA |
| 基金项目:浙江省自然科学基金项目（LQ18H290003） |
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| 中文摘要: |
| 摘 要 目的 探讨大黄素通过调控微小RNA（microRNA）抑制裸鼠SW1990细胞原位移植瘤新生血管的形成及其机制。方法 建立裸鼠SW1990细胞原位移植瘤的动物模型，分为对照组（N组）和3个大黄素组(20mg/kg，40 mg/kg，80 mg/kg，即E20组、E40组、E80组)。各组均采取腹腔注射给药，3次/周，共2周。末次用药后1周处死裸鼠取肿瘤组织。采用免疫组织化学染色法检测肿瘤组织的CD31和CD34（两种不同的内皮细胞标记物），从而确定其微血管密度(MVD)。采用蛋白质印迹法（WesternBlot）法检测新生血管相关基因血管内皮生长因子(VEGF)的表达。荧光定量PCR（RT-qPCR）法检测新生血管相关microRNA miR-20b，miR-21，miR-155，miR-210的表达。结果 免疫组织化学染色显示，按CD31计算的MVD，N组为19.3±1.9，E20组为11.7±1.6，E40组为7.9±2.3，E80组为7.2±1.8，N组、E20组、E40组间两两比较，差异均有统计学意义(P<0.05)；E80组与N组、E20组比较，差异有统计学意义(P<0.05)，与E40比较，差异无统计学意义（P>0.05）。按CD34计算的MVD，N组为16.5±1.1，E20组为12.3±1.8，E40组为5.4±1.6，E80组为4.8±1.6，N组、E20组、E40组间两两比较，差异均有统计学意义(P<0.05)；E80组与N组、E20组比较，差异有统计学意义(P<0.05)，与E40比较，差异无统计学意义（P>0.05）。WesternBlot法结果显示各组VEGF表达分别为1.000±0.054，0.533±0.039，0.381±0.032，0.278±0.031，各组组间两两比较，差异均具有统计学意义(P<0.05)。RT-qPCR法显示各组miR-20b 表达为1.000±0.083，1.398±0.128，1.588±0.140，2.336±0.354，miR-21表达为1.000±0.212，0.449±0.126，0.240±0.051，0.147±0.029， miR-210表达为1.000±0.104，0.378±0.055，0.239±0.034，0.185±0.043， (P<0.05)。各组组间两两比较，差异均具有统计学意义(P<0.05)。miR-155表达为1.000±0.076，0.581±0.128，0.523±0.071，0.402±0.058，N组、E20组、E80组间两两比较，差异均有统计学意义(P<0.05)；E40组与N组、E80组比较，差异有统计学意义(P<0.05)，与E20比较，差异无统计学意义（P>0.05）。结论 大黄素对裸鼠体内人胰腺癌SW1990细胞原位移植瘤的新生血管抑制作用效果显著，其机制可能是大黄素改变新生血管相关基因VEGF的表达以及改变新生血管相关microRNA miR-20b，miR-21，miR-155，miR-210的表达有关。
关键词 大黄素；胰腺癌；新生血管；血管内皮生长因子；微小RNA |
| 英文摘要: |
| Abstract Objective To observe the effect of emodin repressing neovascularization of orthotropic pancreatic cancer nude mouse with SW1990 cell by regulating microRNAs and its mechanism. Methods Build up nude mice animal model by the way of Surgical Orthotropic Implantation (SOI). Dividing into four groups, the control group, 20 mg/kg emodin group, 40 mg/kg emodin group and 80 mg/kg emodin group. The emodin were injected into the abdominal cavity 3 times a week for 2 weeks. The nude mice were sacrificed a week after the process to take the tumor. Detecting the expression of the tumor CD31 and CD34 by the method of immunochemistry, calculate micro vessel density (MVD). Western-Blot were performed to detect the protein expression of tumor neovascularization related factors VEGF. RT-qPCR were performed to detect the neovascularization related microRNAs: miR-20b, miR-21, miR-155 and miR-210. Results Immunochemistry shows the emodin groups’ MVD calculated by CD31 and CD34 is obviously less than the control group, while MVD and emodin dose is negatively related. Western-Blot shows that the emodin is able to decrease the expression of VEGF. RT-qPCR shows the emodin down regulate the microRNAs miR-21, miR-155 and miR-210 expression, meanwhile to raise miR-20b expression. Conclusion Emodin decrease neovascularization of pancreas cancer, its mechanism might be emodin decrease the expression of the angiogenesis-associated factors VEGF and the angiogenesis-associated microRNAs: miR-20b, miR-21, miR-155 and miR-210. |
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