EGFR单抗偶联微管蛋白抑制剂对人肺癌抑制作用的体外研究

发布时间：2018-05-14 22:08

本文选题：抗体药物偶联物(ADC) + 西妥昔单抗　；参考：《南昌大学》2016年硕士论文

【摘要】：目的:本实验拟从体外研究EGFR单抗偶联微管蛋白抑制剂(简称抗体药物偶联物、ADC)对人肺癌A549细胞的抑制作用及作用的细胞周期。方法:1、细胞抑制率:运用CCK-8法检测EGFR单抗偶联微管蛋白抑制剂(ADC)、西妥昔单抗、顺铂在不同浓度不同时间对人肺癌A549细胞的体外抑制作用。2、细胞周期检测:A549细胞在EGFR单抗偶联微管蛋白抑制剂(ADC)、西妥昔单抗、顺铂不同浓度药物作用24小时后,使用流式细胞仪检测细胞周期。结果:1、细胞抑制率结果:ADC组在低、中、高浓度对A549细胞均有明显抑制作用,西妥昔单抗组对A549细胞有一定抑制作用。ADC与西妥昔单抗均为不同浓度时24小时细胞抑制率随着浓度增加而增加,P0.05,有统计学意义。48小时低、中浓度抑制率随着浓度增加而增加,P0.05。到高浓度时不随时间和剂量的增加而增加,P0.05,无统计学意义。ADC与西妥昔单抗两药摩尔剂量接近时,两组在24、48小时比较,均为P0.01,有显著差异。顺铂组低、中、高浓度对A549细胞均有明显抑制作用。低、中浓度时随着浓度和时间增加而增加,P0.05;中浓度与高浓度相同时间比,随着浓度增加,P0.05。高浓度时随着时间延长抑制率无明显增加,P0.05。当ADC摩尔剂量约为顺铂的0.01时,24h抑制率P0.05,无差异;48h抑制率P0.05,ADC作用较顺铂强。提示ADC,西妥昔单抗,顺铂与对照组比较对A549细胞均有抑制作用。在一定剂量范围内ADC与西妥昔单抗、顺铂对A549细胞抑制率均随着浓度和时间的增加而增加,ADC作用明显强于西妥昔单抗。在相同摩尔浓度下ADC作用明显强于顺铂。2、细胞周期结果:ADC在不同浓度组与对照组比较,G2/M期细胞均明显增多,G1和S期细胞均较对照组减少,P0.01,有显著差异,提示ADC为G2/M期阻滞。西妥昔单抗不同浓度以G1期细胞为主,G1、S、G2期与对照组比较均无明显差异P0.05。顺铂组不同浓度细胞各期与对照组比较,G1期细胞明显增多,S、G2期细胞减少,P0.05,有统计学意义,提示为G1期阻滞。相同药物不同浓度之间各期细胞比较P0.05,提示细胞周期的阻滞时期与药物浓度无关。ADC与西妥昔单抗(两药分子量接近)相同摩尔浓度0.0134mmol/L各期比较,P0.01,提示ADC与西妥昔单抗阻滞周期不同。ADC 0.0134mmol/L与顺铂1.33 mmol/L(ADC摩尔数约为顺铂的0.01)各期比较,P0.01为显著差异,提示ADC与顺铂阻滞周期不同。结论:本实验的抗体药物偶联物(EGFR单抗偶联微管蛋白抑制剂)对人肺癌A549细胞生长有明显抑制作用,抑制作用要强于西妥昔单抗与顺铂。EGFR单抗偶联微管蛋白抑制剂阻滞的细胞周期为G2/M期,与西妥昔单抗与顺铂阻滞周期不同。
[Abstract]:Aim: to investigate the inhibitory effect and cell cycle of EGFR monoclonal antibody coupled tubulin inhibitor (EGFR) on human lung cancer A549 cells in vitro. Methods: CCK-8 assay was used to detect EGFR McAb coupled tubulin inhibitor ADCA and cetuximab. The inhibitory effect of cisplatin on human lung cancer A549 cells in vitro was observed at different concentrations and at different times. Cell cycle analysis showed that the cells were treated with EGFR monoclonal antibody coupled with tubulin inhibitor, cetuximab and cisplatin for 24 hours. Cell cycle was measured by flow cytometry. Results the cell inhibition rate of the A549 cells was significantly inhibited in the low, middle and high concentration of the A549 cells in the small, middle and high concentration groups. Cetuximab group had a certain inhibitory effect on A549 cells. When the concentration of ADC and cetuximab were different, the inhibition rate of A549 cells increased with the increase of the concentration of cetuximab. The inhibition rate of A549 cells increased with the increase of concentration, which was significantly lower than that of the control group for 48 hours. The inhibition rate of medium concentration increased with the increase of concentration of A549 cells. At high concentration, there was no increase of P0.05 with the increase of time and dose. When the molar dose of ADC and cetuximab were close to each other, there was a significant difference between the two groups at 24: 48 hours. Low, medium and high concentrations of cisplatin inhibited A549 cells significantly. When the concentration is low and medium concentration increases with the increase of concentration and time, the P0.05 increases with the increase of the concentration and the ratio of the middle concentration to the high concentration is the same as that of the high concentration. At high concentration, the inhibition rate did not increase significantly with time prolongation. When the molar dose of ADC was about 0. 01% of cisplatin, the inhibition rate of P0. 05% was stronger than that of cisplatin at 48 h. These results suggest that ADC, cetuximab and cisplatin can inhibit A549 cells. The inhibitory rates of ADC and cetuximab and cisplatin on A549 cells increased with the increase of concentration and time. At the same molar concentration, the effect of ADC was significantly stronger than that of cisplatin. The cell cycle results showed that the G _ 2 / M phase cells in G _ 2 / M phase were significantly increased in different concentration groups compared with the control group, and the cells in G _ 1 and S phase were significantly lower than those in the control group, indicating that ADC was blocked in G _ 2 / M phase. The concentration of cetuximab in G 1 phase cells was not significantly different from that in control group (P 0.05). Compared with the control group, the cells of different concentrations in cisplatin group increased significantly in G _ 1 phase and decreased P _ (0.05) in S _ (2) G _ 2 phase, indicating that G _ 1 phase was blocked. P0.05 cell comparison between different concentrations of the same drug suggests that the cell cycle arrest period is not related to the concentration of the drug .ADCs are compared with cetuximab (the molecular weight of the two drugs are close) at the same molar concentration of P0.01, suggesting that ADC and cetuximab have the same molecular weight, indicating that the cell cycle arrest period is not related to the concentration of the drug. Compared with cisplatin (0.01 mol number of cisplatin 1.33 mmol/L(ADC), there were significant differences in the block cycles of McAb and cisplatin (P0.01). The results suggest that ADC is different from cisplatin in blocking cycle. Conclusion: the anti-EGFR monoclonal antibody coupled tubulin inhibitor in this study has a significant inhibitory effect on the growth of human lung cancer cell line A549. The inhibitory effect was stronger than that of cetuximab and cisplatin. EGFR monoclonal antibody blocked the cell cycle of G2 / M phase, which was different from that of cetuximab and cisplatin.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R734.2

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