甲磺酸阿帕替尼对人乳腺癌紫杉醇耐药性的逆转作用

Reversal Effect of Apatinib Mesylate on Paclitaxel Resistance of Human Breast Cancer Cells

目的探讨甲磺酸阿帕替尼对人乳腺癌紫杉醇耐药性的逆转作用和机制。方法体外诱导建立人乳腺癌MCF-7/Taxol耐药细胞株,CCK-8法检测阿帕替尼对MCF-7和MCF-7/Taxol细胞株的毒性作用,低毒浓度阿帕替尼联合紫杉醇对MCF-7/Taxol细胞株耐药的逆转作用。实验设置空白对照组(A组),紫杉醇无毒剂量组(B组),阿帕替尼低毒浓度剂量组(C组),紫杉醇无毒剂量联合阿帕替尼低毒浓度剂量组(D组),采用流式细胞术检测MCF-7/Taxol细胞凋亡及细胞周期分布的变化;采用Western Blot法检测P-gp、BCRP、E-cadherin、N-cadherin蛋白表达的变化情况。结果成功构建人乳腺癌MCF-7/Taxol耐药细胞株,阿帕替尼对人乳腺癌MCF-7/Taxol细胞株具有逆转作用。阿帕替尼联合紫杉醇作用于MCF-7/Taxol细胞后与A组、B组、C组比较,细胞凋亡率增加(P<0.05);与A组、B组比较,细胞周期G2/M期分布比例升高(P<0.05);阿帕替尼不影响P-gp、BCRP、E-cadherin和N-cadherin蛋白的表达。结论甲磺酸阿帕替尼具有抑制乳腺癌紫杉醇耐药株的作用,与紫杉醇联合应用协同抑制耐药株细胞并诱导凋亡和细胞周期分布于G2/M期,增强紫杉醇药物敏感性。

Objective To investigate the reversal effect of apatinib mesylate on the paclitaxel resistance of human breast cancer cells(MCF-7/Taxol)and its mechanism.Methods The human breast cancer MCF-7/Taxol resistant cell line was established in vitro.CCK-8 assay was used to detect the toxic effect of apatinib mesylate on MCF-7 and MCF-7/Taxol cell lines,and the reversal effect of low toxic concentration of apatinib mesylate combined with paclitaxel on MCF-7/Taxol cell line drug resistance was detected.The experiment was set up blank control group(group A),paclitaxel non-toxic dose group(group B),apatinib mesylate low toxic dose group(group C),and paclitaxel non-toxic dose combined with apatinib mesylate low toxic dose group(group D).The changes of apoptosis and cell cycle distribution of MCF-7/Taxol cells were detected by flow cytometry.The protein expressions of P-gp,BCRP,E-cadherin and N-cadherin were detected by Western Blot.Results The human breast cancer MCF-7/Taxol resistant cell line was successfully constructed,and apatinib mesylate has a reversal effect on the human breast cancer MCF-7/Taxol cell line.Compared with group A,group B and group C,the apoptosis rate of MCF-7/Taxol cells treated with apatinib mesylate combined with paclitaxel was increased(P<0.05).compared with group A and group B,the proportion of G2/M phase distribution of cell cycle was increased(P<0.05);apatinib mesylate did not affect the protein expression of P-gp,BCRP,E-cadherin and N-cadherin.Conclusion Apatinib mesylate has an inhibitory effect on paclitaxel-resistant breast cancer cells.Apatinib mesylate combined with paclitaxel can synergistically inhibit paclitaxel-resistant breast cancer cells,induce apoptosis and G2/M phase distribution,and enhance the sensitivity to paclitaxel.