奥沙利铂对不同转移潜能的人结肠癌细胞株增殖和凋亡的影响

发布时间：2018-01-24 05:00

本文关键词： 奥沙利铂 SW480细胞 SW620细胞细胞周期细胞凋亡　出处：《兰州大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的探讨奥沙利铂(L-OHP)体外对人结肠癌细胞SW480和SW620增殖和凋亡的影响及其可能机制。方法1.MTT比色法检测用不同质量浓度(2、4、8、16、32和64μg/m L)L-OHP与SW480和SW620细胞作用24、48和72 h后对增殖抑制作用的影响。2.DAPI染色观察经不同质量浓度(4、16和64μg/m L)L-OHP处理24 h后细胞核形态的变化。3.流式细胞术检测用不同质量浓度(1/2IC50和IC50)L-OHP处理24 h后SW480和SW620细胞周期和凋亡率。4.Western blot法检测经不同质量浓度(4、16和64μg/mL)L-OHP处理24 h后SW480和SW620细胞中凋亡相关蛋白Bax和Caspaes-3蛋白的表达水平以及DNA损伤相关蛋白γHA2X蛋白的表达水平。结果1.L-OHP对SW480和SW620细胞的增殖有明显的抑制作用且呈时间-剂量依赖性(P0.05)。SW620细胞对L-OHP的敏感性明显高于SW480细胞(P0.05)。2.L-OHP作用于SW480和SW620细胞24 h后,随着药物浓度的增加正常细胞数目减少,出现细胞核固缩、多核、核碎裂及凋亡小体的细胞数目明显增加(P0.05)。相同浓度L-OHP作用下,SW620细胞中出现细胞核固缩、多核、核碎裂及凋亡小体的细胞的比例明显较SW480细胞中高(P0.05)。3.L-OHP对SW480和SW620细胞的有明显的促进早期凋亡的作用且呈时间-剂量依赖性(P0.05)。相同浓度L-OHP作用下对SW620细胞促进早期凋亡的作用明显大于SW480细胞(P0.05)。4.自然对数增殖周期,SW620细胞S期细胞比例低于SW480细胞,而G1期细胞比例高于SW480细胞(P0.05)。用不同质量浓度的L-OHP(1/2IC50、IC50)作用24 h,SW480细胞G1期细胞比例随浓度增高而显著降低,G2/M期细胞比例显著升高(P0.05),SW620细胞G1期细胞比例随浓度增高而显著降低,S期细胞比例显著升高(P0.05)。5.L-OHP作用于SW480和SW620细胞24 h后,随着药物浓度的增加SW480和SW620细胞中凋亡相关蛋白Bax和Caspase-3蛋白以及DNA损伤相关蛋白γHA2X蛋白的表达量明显增加(P0.05)。相同药物浓度下SW620细胞中凋亡相关蛋白Bax和Caspase-3蛋白以及DNA损伤相关蛋白γHA2X蛋白的表达量明显较SW480高(P0.05)。结论1.L-OHP对二种结肠癌细胞株都有明显的增殖抑制作用,且对SW620细胞的抑制作用明显大于SW480细胞。2.L-OHP主要通过阻滞细胞于G2/M期或S期抑制二种肿瘤细胞增殖并诱导细胞凋亡。3.L-OHP诱导二种肿瘤细胞凋亡的机制可能是通过上调促凋亡蛋白Bax和启动Caspaes级联反应来实现的。DNA损伤可能也是其促进凋亡的机制之一。但二者之间的具体联系还需进一步实验来探讨。
[Abstract]:Objective to investigate the effect of oxaliplatin L-OHPin on proliferation and apoptosis of human colon cancer cell line SW480 and SW620 in vitro and its possible mechanism. 2. (4) the cells of SW480 and SW620 were exposed to L-OHP at the level of 64 渭 g / L and 32 渭 g / m ~ (-1), 24 渭 g / m ~ (-1), respectively. Effects of 48 h and 72 h on proliferation inhibition. 2. DAPI staining was performed at different concentrations. Changes of nuclear morphology after 24 h treatment with 16 and 64 渭 g / m L LL-OHP .3. flow cytometry was used to detect nuclear morphology with different mass concentrations of 1 / 2 IC50 and IC50). After treatment with L-OHP for 24 h, the cell cycle and apoptosis rate of SW480 and SW620 were detected by Western blot assay. 16 and 64 渭 g / mL). L-OHP treatment 24. The expression levels of apoptosis-related proteins (Bax and Caspaes-3) and DNA damage associated protein 纬 HA2X (纬 HA2X) were detected in SW480 and SW620 cells for 5 h. Results 1. L-OHP inhibited the proliferation of SW480 and SW620 cells in a time-dose dependent manner. The sensitivity of P0.05N. SW620 cells to L-OHP was significantly higher than that of SW480 cells (P0.05). 2.L-OHP treated SW480 and SW620 cells for 24 h. With the increase of drug concentration, the number of normal cells decreased, the number of cells with nuclear pyknosis, polynuclear, nuclear fragmentation and apoptotic bodies increased significantly. The nuclei of SW620 cells were pyknotic and polynuclear. The percentage of cells with nuclear fragmentation and apoptotic bodies was significantly higher than that of SW480 cells (P 0.05). 3. L-OHP can promote early apoptosis in SW480 and SW620 cells in a time-dose dependent manner (P0.05). The effect of L-OHP at the same concentration on the early apoptosis of SW620 cells was significantly greater than that on SW480 cells (P 0.05N. 4). The natural logarithmic proliferative cycle was observed. The percentage of S phase cells of SW620 cells was lower than that of SW480 cells, while the proportion of G1 phase cells was higher than that of SW480 cells. The ratio of G _ 1 phase cells in G _ 2 / M phase decreased significantly with the increase of the concentration of IC50), and the percentage of G _ 1 phase cells increased significantly (P 0.05). The proportion of SW620 cells in G1 phase decreased significantly with the increase of concentration. The proportion of S phase cells increased significantly (P 0.05). 5. L-OHP treated SW480 and SW620 cells for 24 h. With the increase of drug concentration, the expression of apoptosis-related proteins (Bax and Caspase-3) and DNA damage associated protein 纬 HA2X (纬 HA2X) in SW480 and SW620 cells increased significantly. (. P0.05). The expression of apoptosis-related proteins Bax, Caspase-3 and DNA damage associated protein 纬 HA2X in SW620 cells was significantly higher than that in SW480 at the same drug concentration (P < 0.05). Conclusion 1. L-OHP can inhibit the proliferation of two colon cancer cell lines. Moreover, the inhibitory effect of L-OHP on SW620 cells was significantly greater than that on SW480 cells. 2.L-OHP inhibited the proliferation of two kinds of tumor cells and induced apoptosis mainly by blocking the cells in G _ 2 / M phase or S phase. The mechanism of apoptosis induced by L-OHP may be through up-regulation of apoptosis-promoting protein Bax and initiation of Caspaes cascade reaction. 1. But the specific connection between the two needs further experiment to explore.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.3

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