紫杉醇耐药对食管癌细胞增殖、迁移及血管形成的影响

Effects of paclitaxel resistance on proliferation,migration and angiogenesis of esophageal cancer cells

目的研究紫杉醇(paclitaxel,PTX)耐药对食管癌细胞增殖、迁移及血管形成的影响。方法采用体外间歇诱导加逐步递增PTX浓度的方法建立PTX耐药细胞系KYSE150/PTX。倒置显微镜下观察PTX耐药细胞形态变化,MTT法检测PTX耐药对食管癌细胞细胞增殖能力的影响,克隆形成试验观察PTX耐药对食管癌细胞克隆形成能力的影响,划痕愈合试验检测PTX耐药对食管癌细胞迁移能力的影响,体外小管形成试验检测PTX耐药对食管癌细胞体外小管形成能力的影响。结果KYSE150细胞最终能在含0.05 mg/L PTX的培养基中正常生长,此细胞即为PTX耐药细胞系KYSE150/PTX。KYSE150细胞对PTX产生稳定耐药后,显微镜下未见细胞形态上的改变;与KYSE150细胞相比,KYSE150/PTX细胞的增殖能力及克隆形成能力明显减弱(P<0.05),划痕愈合能力增强(P<0.001),体外小管形成数目增加(P<0.001)。结论经过10个月成功构建了PTX耐药细胞。PTX耐药导致食管癌细胞增殖能力下降、迁移及体外血管形成能力增加,可能是PTX耐药影响临床治疗效果的重要因素。此外,耐药模型的建立为深入探讨食管癌化疗耐药作用及其机制奠定了良好的实验基础。

Objective To investigate the effect of paclitaxel(PTX)resistance to the proliferation,migration and angiogenesis of esophageal cancer cells.Methods PTX-resistant KYSE150/PTX cell line was established by low-dose intermittent induction and gradually increasing PTX concentration.The morphology of KYSE150/PTX cells was observed under inverted microscope.The effect of PTX resistance on the proliferation of esophageal cancer cells was observed by MTT assay,while that on colony formation by colony formation assay,that on migration by wound healing assay,and that on angiogenesis by in vitro tubule formation assay.Results The KYSE150/PTX cells growing normally in the medium containing 0.05 mg/L PTX was served as PTX-resistant KYSE150/PTX,which showed no change in morphology under microscope.Compared with those of KYSE150 cells,the abilities of KYSE150/PTX cells in proliferation and colony formation were reduced(P<0.05),while that in wound healing and tubule formation ability were enhanced(each P<0.001).Conclusion PTX-resistant KYSE150/PTX cell line was successfully established within 10 months.PTX resistance might influence the therapeutic effect by inhibiting the proliferation ability and enhancing the migration and angiogenesis abilities of human esophageal cancer cells.In addition,the establishment of a drug resistance model provides a good experimental basis for further exploring the mechanism of chemotherapy resistance of esophageal cancer.