微管蛋白抑制剂ZP-20f对口腔鳞状细胞癌周期及凋亡的作用机制
发布时间：2019-02-19 20:04
【摘要】：目的:本实验采用MTT实验,Western Blot实验,平板克隆实验及流式细胞实验检测微管蛋白抑制剂ZP-20f对体外培养的口腔鳞癌细胞的增殖、凋亡及细胞周期的影响,并进行相关数据分析。探讨微管蛋白抑制剂Zp-20f在口腔鳞状细胞癌中的作用机制。方法:1、口腔鳞癌SCC-9、CAL-27细胞的细胞培养。2、MTT法检测Zp-20f对口腔鳞癌细胞SCC-9、CAL-27细胞的毒性作用,并计算最实试验浓度IC50。3、流式细胞术检测口腔鳞癌细胞SCC-9、CAL-27细胞的周期及凋亡率。4、应用平板克隆形成实验分析Zp-20f对口腔鳞状细胞癌细胞SCC-9、CAL-27的毒性效应。5、Wetern Blot检测SCC-9、CAL-27细胞中相关蛋白的表达变化。结果:1、ZP-20f可显著抑制口腔鳞癌细胞增殖ZP-20f在一定浓度范围内能显著抑制口腔鳞癌的增殖作用,且其抑制活性具有浓度依赖作用。平板克隆实验可以看出ZP-20f能浓度依赖的抑制肿瘤细胞克隆的形成。2、ZP-20f可在G2/M期阻滞细胞周期ZP-20f在一定浓度范围内能显著诱导口腔鳞癌细胞G2/M期阻滞,并具随着化合物浓度的增加,G2/M期细胞比例随之增加。3、ZP-20f可在G2/M期阻滞细胞周期ZP-20f在一定浓度范围内能显著诱导口腔鳞癌细胞G2/M期阻滞,并具随着化合物浓度的增加,G2/M期细胞比例随之增加。4、细胞周期相关蛋白量发生变化化合物在一定浓度范围内能引起细胞内周期和凋亡相关蛋白含量的变化,使细胞中周期与凋亡相关蛋白Cycling B1、p-H3和cleaved-PARP的量显著增加,且具有浓度依赖性(图5)。结论:1、ZP-20f对口腔鳞癌细胞SCC-9和Cal-27的体外增殖抑制活性较强,能明显将细胞周期阻滞在G2/M期,并能诱导细胞凋亡;2、ZP-20f在体外有较好的抗肿瘤作用。其作用机制可能与p-H3或cleaved-PARP的上调及对微管蛋白的抑制作用相关,是一个一个非常有前景的抗肿瘤药物。
[Abstract]:Objective: to investigate the effects of tubulin inhibitor (ZP-20f) on the proliferation, apoptosis and cell cycle of oral squamous carcinoma cells cultured in vitro by MTT, Western Blot assay, plate clone assay and flow cytometry. And carries on the correlation data analysis. To investigate the mechanism of tubulin inhibitor Zp-20f in oral squamous cell carcinoma. Methods: 1. The cell culture of oral squamous cell carcinoma (SCC-9,CAL-27) cells. 2MTT assay was used to detect the toxic effect of Zp-20f on oral squamous cell carcinoma SCC-9,CAL-27 cells, and the concentration of IC50.3, was calculated. Flow cytometry was used to detect the cell cycle and apoptosis rate of oral squamous cell SCC-9,CAL-27 cells. 4. The toxic effect of Zp-20f on SCC-9,CAL-27 of oral squamous cell carcinoma cells was analyzed by plate clone formation assay. The expression of related proteins in SCC-9,CAL-27 cells was detected by Wetern Blot. Results: 1ZP-20f could significantly inhibit the proliferation of oral squamous cell carcinoma (OSCC) cell line ZP-20f in a certain concentration range, and its inhibitory activity was concentration-dependent. Plate cloning assay showed that ZP-20f could inhibit the formation of tumor cell clone in a concentration-dependent manner. 2ZP-20f could significantly induce G _ 2 / M phase arrest of oral squamous carcinoma cells in a certain concentration range by blocking cell cycle ZP-20f in G _ 2 / M phase. With the increase of the concentration of compounds, the proportion of G2 / M phase cells increased. 3ZP-20f could significantly induce the G _ 2 / M phase arrest of oral squamous carcinoma cells in a certain concentration range by blocking cell cycle ZP-20f in G _ 2 / M phase. With the increase of the concentration of compounds, the proportion of cells in G _ 2 / M phase increased. 4. The change of cell cycle associated protein content in a certain concentration range could cause changes in cell cycle and apoptosis-related protein content. Cell cycle and apoptosis-related protein Cycling B1, p-H3 and cleaved-PARP were significantly increased in a concentration-dependent manner (Fig. 5). Conclusion: (1) ZP-20f can inhibit the proliferation of oral squamous cell SCC-9 and Cal-27 in vitro, block the cell cycle at G _ 2 / M phase and induce apoptosis, and ZP-20f has a good anti-tumor effect in vitro. The mechanism may be related to the upregulation of p-H3 or cleaved-PARP and the inhibition of tubulin. It is a promising antitumor drug.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.8
本文编号：2426827
[Abstract]:Objective: to investigate the effects of tubulin inhibitor (ZP-20f) on the proliferation, apoptosis and cell cycle of oral squamous carcinoma cells cultured in vitro by MTT, Western Blot assay, plate clone assay and flow cytometry. And carries on the correlation data analysis. To investigate the mechanism of tubulin inhibitor Zp-20f in oral squamous cell carcinoma. Methods: 1. The cell culture of oral squamous cell carcinoma (SCC-9,CAL-27) cells. 2MTT assay was used to detect the toxic effect of Zp-20f on oral squamous cell carcinoma SCC-9,CAL-27 cells, and the concentration of IC50.3, was calculated. Flow cytometry was used to detect the cell cycle and apoptosis rate of oral squamous cell SCC-9,CAL-27 cells. 4. The toxic effect of Zp-20f on SCC-9,CAL-27 of oral squamous cell carcinoma cells was analyzed by plate clone formation assay. The expression of related proteins in SCC-9,CAL-27 cells was detected by Wetern Blot. Results: 1ZP-20f could significantly inhibit the proliferation of oral squamous cell carcinoma (OSCC) cell line ZP-20f in a certain concentration range, and its inhibitory activity was concentration-dependent. Plate cloning assay showed that ZP-20f could inhibit the formation of tumor cell clone in a concentration-dependent manner. 2ZP-20f could significantly induce G _ 2 / M phase arrest of oral squamous carcinoma cells in a certain concentration range by blocking cell cycle ZP-20f in G _ 2 / M phase. With the increase of the concentration of compounds, the proportion of G2 / M phase cells increased. 3ZP-20f could significantly induce the G _ 2 / M phase arrest of oral squamous carcinoma cells in a certain concentration range by blocking cell cycle ZP-20f in G _ 2 / M phase. With the increase of the concentration of compounds, the proportion of cells in G _ 2 / M phase increased. 4. The change of cell cycle associated protein content in a certain concentration range could cause changes in cell cycle and apoptosis-related protein content. Cell cycle and apoptosis-related protein Cycling B1, p-H3 and cleaved-PARP were significantly increased in a concentration-dependent manner (Fig. 5). Conclusion: (1) ZP-20f can inhibit the proliferation of oral squamous cell SCC-9 and Cal-27 in vitro, block the cell cycle at G _ 2 / M phase and induce apoptosis, and ZP-20f has a good anti-tumor effect in vitro. The mechanism may be related to the upregulation of p-H3 or cleaved-PARP and the inhibition of tubulin. It is a promising antitumor drug.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.8
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