大黄素衍生物A抑制卵巢癌淋巴结高转移细胞的作用及机制研究
发布时间：2018-12-10 07:04
【摘要】：第一章大黄素衍生物A对SKOV3和SKOV3-pm4细胞迁移侵袭能力影响目的研究大黄素衍生物A对具有不同淋巴结转移潜能的卵巢癌细胞的迁移和侵袭能力的抑制作用。方法四甲基偶氮唑蓝(MTT)法检测不同浓度大黄素衍生物A处理人卵巢浆液性乳头状腺癌细胞(SKO V3)和具有淋巴结定向高转移能力的亚克隆细胞(SKOV3-pm4) 24h后细胞的增殖作用。用无细胞毒性浓度的药物作用两种细胞后,用细胞划痕实验检测两种细胞的迁移能力,用Transwell小室侵袭实验测定细胞的侵袭能力。结果在一定浓度范围内大黄素衍生物A对SKOV3和SKOV3-pm4细胞的增殖均有抑制作用,24h的半数抑制浓度(IC50)分别为23.21mg/L、34.46mg/L细胞划痕实验结果显示大黄素衍生物A处理后两种细胞的迁移能力减弱。Transwell小室侵袭实验结果显示,大黄素衍生物A处理后两种细胞的侵袭能力均受到抑制。结论大黄素衍生物A能抑制具有高淋巴结转移潜能的卵巢癌细胞的迁移和侵袭能力。第二章细胞中Rac1/Cdc42蛋白的分布以及大黄素衍生物A对Rac1/Cdc42基因和蛋白表达的影响目的探讨大黄素衍生物A抑制具有不同淋巴结转移潜能的卵巢癌细胞的迁移和侵袭能力的可能的作用机制。方法免疫荧光法观察两种细胞中Racl和Cdc42蛋白的定位和分布情况。用无细胞毒性浓度的药物作用两种细胞后,采用实时荧光定量PCR (RT-PC R)检测Rac1、Cdc42基因的表达情况,用蛋白免疫印迹法(Western Blot)检测Rac1、Cdc42蛋白的表达水平。结果免疫荧光法观察到两种细胞中Rac1和Cdc42蛋白均广泛分布于细胞质中,Rac1蛋白在细胞核周围比较密集。RT-PCR和Western Blot检测结果显示随着药物浓度2 mg/L、4mg/L、8 mg/L的梯度增加,Racl基因和蛋白的表达水平均降低,与空白对照组相比,差异有统计学意义(P0.05)。而Cdc42基因和蛋白的表达变化不明显。结论大黄素衍生物A抑制具有高淋巴结转移潜能的卵巢癌细胞的迁移和侵袭能力的作用机制可能与下调Rac1蛋白的表达有关。
[Abstract]:Chapter 1 effects of emodin derivative A on migration and invasion of SKOV3 and SKOV3-pm4 cells objective to study the inhibitory effect of emodin derivative A on migration and invasion of ovarian cancer cells with different lymph node metastasis potential. Methods Human ovarian serous papillary adenocarcinoma cells (SKO V3) and subclone cells (SKOV3-pm4) with high lymph node metastasis were detected by (MTT) assay with different concentrations of emodin derivative A. The proliferation of posterior cells. After two kinds of cells were treated with drugs without cytotoxic concentration, the migration ability of two kinds of cells was detected by cell scratch assay, and the invasion ability of two kinds of cells was measured by Transwell chamber invasion test. Results in a certain concentration range, emodin derivative A inhibited the proliferation of SKOV3 and SKOV3-pm4 cells, and the half inhibitory concentration (IC50) at 24 h was 23.21 mg / L, respectively. The results of 34.46mg/L cell scratch test showed that the migration ability of the two kinds of cells was weakened after treatment with emodin derivative A, and the invasion ability of the two kinds of cells was inhibited by Transwell chamber invasion assay. Conclusion emodin derivative A can inhibit the migration and invasion of ovarian cancer cells with high lymph node metastasis potential. The distribution of Rac1/Cdc42 protein in cells and the effect of emodin derivative A on the expression of Rac1/Cdc42 gene and protein objective to investigate the inhibitory effect of emodin derivative A on migration of ovarian cancer cells with different lymph node metastasis potential And the possible mechanism of invasion ability. Methods the localization and distribution of Racl and Cdc42 proteins were observed by immunofluorescence. The expression of Rac1,Cdc42 gene was detected by real-time fluorescence quantitative PCR (RT-PC R), and the expression of Rac1,Cdc42 protein was detected by Western blot (Western Blot). Results the Rac1 and Cdc42 proteins were widely distributed in the cytoplasm of the two cells by immunofluorescence assay, and the Rac1 protein was dense around the nucleus. The results of RT-PCR and Western Blot detection showed that with the concentration of 2 mg/L,4mg/L,, 8 the gradient of mg/L increased and the expression of Racl gene and protein decreased, compared with the control group, the difference was statistically significant (P0.05). However, the expression of Cdc42 gene and protein did not change significantly. Conclusion the mechanism of emodin derivative A in inhibiting the migration and invasion of ovarian cancer cells with high lymph node metastasis potential may be related to the down-regulation of Rac1 protein expression.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R737.31
本文编号：2370168
[Abstract]:Chapter 1 effects of emodin derivative A on migration and invasion of SKOV3 and SKOV3-pm4 cells objective to study the inhibitory effect of emodin derivative A on migration and invasion of ovarian cancer cells with different lymph node metastasis potential. Methods Human ovarian serous papillary adenocarcinoma cells (SKO V3) and subclone cells (SKOV3-pm4) with high lymph node metastasis were detected by (MTT) assay with different concentrations of emodin derivative A. The proliferation of posterior cells. After two kinds of cells were treated with drugs without cytotoxic concentration, the migration ability of two kinds of cells was detected by cell scratch assay, and the invasion ability of two kinds of cells was measured by Transwell chamber invasion test. Results in a certain concentration range, emodin derivative A inhibited the proliferation of SKOV3 and SKOV3-pm4 cells, and the half inhibitory concentration (IC50) at 24 h was 23.21 mg / L, respectively. The results of 34.46mg/L cell scratch test showed that the migration ability of the two kinds of cells was weakened after treatment with emodin derivative A, and the invasion ability of the two kinds of cells was inhibited by Transwell chamber invasion assay. Conclusion emodin derivative A can inhibit the migration and invasion of ovarian cancer cells with high lymph node metastasis potential. The distribution of Rac1/Cdc42 protein in cells and the effect of emodin derivative A on the expression of Rac1/Cdc42 gene and protein objective to investigate the inhibitory effect of emodin derivative A on migration of ovarian cancer cells with different lymph node metastasis potential And the possible mechanism of invasion ability. Methods the localization and distribution of Racl and Cdc42 proteins were observed by immunofluorescence. The expression of Rac1,Cdc42 gene was detected by real-time fluorescence quantitative PCR (RT-PC R), and the expression of Rac1,Cdc42 protein was detected by Western blot (Western Blot). Results the Rac1 and Cdc42 proteins were widely distributed in the cytoplasm of the two cells by immunofluorescence assay, and the Rac1 protein was dense around the nucleus. The results of RT-PCR and Western Blot detection showed that with the concentration of 2 mg/L,4mg/L,, 8 the gradient of mg/L increased and the expression of Racl gene and protein decreased, compared with the control group, the difference was statistically significant (P0.05). However, the expression of Cdc42 gene and protein did not change significantly. Conclusion the mechanism of emodin derivative A in inhibiting the migration and invasion of ovarian cancer cells with high lymph node metastasis potential may be related to the down-regulation of Rac1 protein expression.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R737.31
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