顺铂耐药对食管癌细胞增殖、凋亡、迁移和血管生成的影响

发布时间：2019-03-22 20:42

【摘要】：目的:探讨顺铂(cis-dichlorodiamine platinum,cDDP)耐药对人食管癌KYSE150细胞增殖、凋亡、迁移和血管生成的影响。方法:首先采用顺铂浓度递增的方法,历经10个月左右的时间成功建立了人食管癌顺铂耐药细胞系KYSE150/cDDP。采用MTT法测定其药物敏感性,并通过形态学观察、MTT实验、集落形成实验、DAPI染色、划痕愈合实验及小管形成实验比较顺铂耐药后食管癌细胞生物学行为的改变。结果:KYSE150细胞和KYSE150/cDDP细胞在形态上无明显差异;MTT实验结果显示,与KYSE150细胞相比,KYSE150/cDDP细胞对cDDP的耐药指数为6.35,细胞活力下降;集落形成实验结果显示,KYSE150/cDDP细胞的集落形成率为(15.00±3.05)%,而KYSE150细胞的集落形成率为(86.70±6.57)%;DAPI染色显示KYSE150/cDDP细胞的凋亡率为(0.63±0.09)%,而KYSE150细胞的凋亡率为(8.46±1.33)%;划痕愈合实验显示,与KYSE150细胞相比,KYSE150/cDDP细胞的划痕愈合能力更强,其迁移率较KYSE150细胞的迁移率高;小管形成实验显示,KYSE150/cDDP细胞的血管生成数为76.20±3.18,而KYSE150细胞的血管生成数为50.60±1.33;Western blot实验结果显示KYSE150/cDDP细胞中MMP-2和VEGFR2的表达均高于KYSE150细胞。结论:KYSE150/cDDP细胞具有耐药表型,耐药后生长缓慢,凋亡能力降低,迁移和血管生成能力增加,这可能是临床化疗失败的重要原因。
[Abstract]:Aim: to investigate the effects of cisplatin (cis-dichlorodiamine platinum,cDDP) resistance on proliferation, apoptosis, migration and angiogenesis of human esophageal carcinoma KYSE150 cells. Methods: the cisplatin resistant human esophageal cancer cell line KYSE150/cDDP. was successfully established after 10 months by increasing the concentration of cisplatin. The drug sensitivity was measured by MTT method. The changes of biological behavior of esophageal cancer cells after cisplatin resistance were compared by morphological observation, MTT test, colony formation test, DAPI staining, scratch healing test and tubule formation test. Results: there was no significant difference between KYSE150 cells and KYSE150/cDDP cells in morphology, MTT assay showed that compared with KYSE150 cells, the drug resistance index of KYSE150/cDDP cells to cDDP was 6.35, and the viability of KYSE150/cDDP cells decreased. The colony forming rate of KYSE150/cDDP cells was (15.00 卤3.05)%, while that of KYSE150 cells was (86.70 卤6.57)%. The apoptosis rate of KYSE150/cDDP cells was (0.63 卤0.09)% and that of KYSE150 cells was (8.46 卤1.33)% by DAPI staining. Scratch healing test showed that the ability of scratch healing of KYSE150/cDDP cells was stronger than that of KYSE150 cells, and the migration rate of KYSE150/cDDP cells was higher than that of KYSE150 cells. Tubule formation assay showed that the angiogenesis number of KYSE150/cDDP cells was 76.20 卤3.18, while that of KYSE150 cells was 50.60 卤1.33%. Western blot assay showed that the expression of MMP-2 and VEGFR2 in KYSE150/cDDP cells was higher than that in KYSE150 cells. Conclusion: KYSE150/cDDP cells have drug-resistant phenotype, slow growth, decreased apoptosis, and increased migration and angiogenesis ability after drug resistance, which may be an important reason for the failure of clinical chemotherapy.
【作者单位】：山西医科大学生物化学与分子生物学教研室;
【基金】：国家自然科学基金资助项目(No.81372676) 中国博士后基金资助项目(No.2014M551058) 山西省自然科学基金资助项目(No.201601D011130)
【分类号】：R735.1

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