中文摘要：

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。

英文摘要：

AIM：To investigate the effect of cis-dichlorodiamine platinun（cDDP）resistance on proliferation,apoptosis,migration and angiogenesis of esophageal cancer cell line KYSE150. METHODS：Using the method of increasing concentration of cDDP in culture for 10 months,the human esophageal carcinoma cDDP-resistant cell line named KYSE150/cDDP was established successfully. The drug sensitivity was measured by MTT assay. The changes of the biological behaviors between the parental cell line and resistant cell line were determined by morphological observation assay,MTT assay,colony formation assay,DAPI staining,wound healing assay and tube formation experiment. RESULTS：No significant morphological difference between KYSE150 cells and KYSE150/cDDP cells was observed. Compared with KYSE150 cells,the drug resistance index of KYSE150/cDDP cells was 6. 35,and the viability of KYSE150/cDDP cells was decreased. The colony formation rate of KYSE150/cDDP cells was（15. 00 ± 3. 05）%,while the colony formation rate of KYSE150 cells was（86. 70 ± 6. 57）%. The apoptotic rate of KYSE150/cDDP cells was（0. 63 ± 0. 09）%,and that of KYSE150 cells was（8. 46 ± 1. 33）%. Compared with KYSE150 cells,KYSE150/cDDP cells showed a stronger healing ability of scratch,and the migration rate was higher than that of KYSE150 cells. The results of tube formation experiment showed that the vessel number in KYSE150/cDDP group was 76. 20 ± 3. 18,while the vessel number in KYSE150 group was 50. 60 ± 1. 33. The protein expression of MMP-2 and VEGFR2 in KYSE150/cDDP cells was higher than that in KYSE150 cells. CONCLUSION：KYSE150/cDDP cells present drug-resistant phenotype and show a slow growth rate.The ability of apoptosis is decreased,and the abilities of cell migration and angiogenesis are increased. This may be an important reason for the failure of clinical chemotherapy for esophageal cancer.