三七姜挥发油对人鼻咽癌CNE-2细胞的侵袭和迁移抑制作用及其机制初步探讨

发布时间：2018-05-23 22:37

本文选题：三七姜挥发油 + CNE-2细胞　；参考：《桂林医学院》2017年硕士论文

【摘要】：目的:目的是研究三七姜挥发油对人鼻咽癌CNE-2细胞侵袭和迁移的抑制作用,并探讨其可能机制。方法:体外培养人鼻咽癌CNE-2细胞,CCK8实验检测三七姜挥发油对CNE-2细胞增值能力的抑制作用,计算生长抑制率及半数抑制浓度(IC50);选取三七姜挥发油(0,20,40,60μg·ml-1)作用于CNE-2细胞0,24,48h,分别于三个不同时间段,采用划痕实验检测三七姜挥发油对CNE-2细胞迁移能力的抑制作用;Transwell实验检测三七姜挥发油对CNE-2细胞侵袭能力的抑制作用。取作用48h后的CNE-2细胞,采用蛋白印迹法检测MMP-2、MMP-9和VEGF等侵袭和迁移相关基因蛋白水平的表达;RT-PCR技术检测MMP-2、MMP-9和VEGF等相关侵袭和迁移基因的表达情况。结果:三七姜挥发油作用于CNE-2细胞24h,48h,72h后,细胞半数抑制浓度分别为70.296μg·ml-1,63.749μg·ml-1,59.532μg·ml-1,呈时间-剂量依赖性(P0.05);不同浓度的三七姜挥发油(0,20,40,60μg·ml-1)作用于细胞48h后,划痕实验发现与空白对照组相比,细胞的迁移能力随着药物浓度的增加而降低;Transwell实验显示,随着药物浓度逐渐的增加,细胞的侵袭能力逐渐下降;RT-PCR显示,与阴性对照组相比,MMP-2、MMP-9和VEGF等基因表达下调,且变化趋势呈一定的浓度依赖性。蛋白免疫印迹法显示,与阴性对照组相比,MMP-2、MMP-9和VEGF等蛋白表达下调,且变化趋势呈一定的浓度依赖性。结论:三七姜挥发油能抑制人鼻咽癌CNE-2细胞侵袭和迁移的能力而发挥抗肿瘤的作用,并且成浓度依赖性,其机制可能与通过下调MMP-2、MMP-9和VEGF等基因的表达,进而抑制鼻咽癌肿瘤细胞的转移有关。
[Abstract]:Aim: to study the inhibitory effect of volatile oil of Panax notoginseng on invasion and migration of human nasopharyngeal carcinoma (NPC) CNE-2 cells and its possible mechanism. Methods: the inhibitory effect of essential oil of Panax notoginseng on the proliferation of human nasopharyngeal carcinoma (NPC) CNE-2 cells was detected by CCK8 assay. The growth inhibition rate and the half inhibitory concentration (IC50) were calculated, and the essential oil of Radix Notoginseng was selected. The inhibitory effect of essential oil of Panax notoginseng on the migration of CNE-2 cells was detected by scratch test. The inhibitory effect of volatile oil of Panax notoginseng on the invasion of CNE-2 cells was detected by Transwell test. The expression of MMP-2, MMP-9, VEGF and other invasion and migration-related genes were detected by Western blotting in CNE-2 cells after 48 h. RT-PCR was used to detect the expression of invasion and migration genes such as MMP-2, MMP-9, VEGF and so on. Results: after treated with the volatile oil of Panax notoginseng for 24 h and 48 h and 72 h after treatment, the cell half inhibitory concentrations were 70.296 渭 g ml-1nl and 63.749 渭 g ml-1n 59.532 渭 g ml-1, respectively, in a time-dose dependent manner (P0.05A), and different concentrations of volatile oil 020404060 渭 g ml-1) were applied to the cells for 48 h. The migration ability of cells decreased with the increase of drug concentration. With the increase of drug concentration, the invasiveness of cells decreased gradually. Compared with negative control group, the expression of MMP-2, MMP-9, VEGF and other genes were down-regulated by reverse transcription-polymerase chain reaction (RT-PCR). The trend of change was in a concentration dependent manner. Western blot showed that the expression of MMP-2, MMP-9 and VEGF were down-regulated in a concentration dependent manner. Conclusion: the essential oil of Panax notoginseng can inhibit the invasion and migration of human nasopharyngeal carcinoma CNE-2 cells and play an antitumor role in a concentration-dependent manner. The mechanism may be related to the down-regulation of the expression of MMP-2, MMP-9 and VEGF genes. And then inhibit the metastasis of nasopharyngeal carcinoma tumor cells.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.63

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