PCDH10减少多发性骨髓瘤细胞逃避表阿霉素杀伤

PCDH10 prevents multiple myeloma cell escape from epirubicin-induced cytotoxicity

目的:探讨原钙黏蛋白10(protocadherin-10,PCDH10)对表阿霉素杀伤多发性骨髓瘤(multiple myeloma,MM)细胞的影响及机制。方法:采用脂质体法将PCDH10基因转染入人MM细胞株RPMI8226细胞,RT-PCR和Western blot法检测PCDH10基因和蛋白的表达;CCK-8法检测表阿霉素对RPMI8226细胞的增殖抑制作用,求得半数抑制浓度(half-maximal inhibitory concentration,IC50);Transwell法检测表阿霉素和PCDH10对RPMI8226细胞迁移的影响;流式细胞术检测CXC族趋化因子受体4(CXC chemokine receptor 4,CXCR4)蛋白表达;Western blot方法检测Ras同源家族成员A(ras homolog gene family member A,Rho A)和基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)蛋白表达情况。结果:PCDH10基因成功转染入RPMI8226细胞,并成功表达;PCDH10转染组表阿霉素IC50为(0.87±0.07)μg/ml,明显低于空质粒转染组(1.27±0.02)μg/ml(F=77.93,P=0.000);PCDH10减少表阿霉素所致的细胞迁移(F=34.943,P=0.000)并抑制CXCR4(F=117.269,P=0.000)、Rho A(F=1 818.250,P=0.000)和MMP-9(F=523.764,P=0.000)蛋白表达。结论:PCDH10一方面可增强MM细胞对表阿霉素的敏感性,另一方面可通过抑制CXCR4、Rho A、MMP-9蛋白的表达来减少MM细胞逃避表阿霉素的杀伤,从而增强了表阿霉素的抗骨髓瘤效应。

Objective：To explore the influence and mechanism of PCDH10 on the chemotherapy of epirubicin in multiple myeloma（MM）. Methods：The PCDH10 expression was regulated by transfecting pc DNA3.1（＋）PCDH10 in RPMI8226 cells. RT-PCR and Western blot were used to test the transfection effect. The CCK-8 assay was used to evaluate cell proliferation. The Boyden chamber and matrigel glue were used to construct migration and invasion model for detecting the influence of PCDH10 on epirubicin-induced cell escape. Flow cytometry was used to test the CXCR4 protein expression. Western blot was used to assess the expression of Rho A and MMP-9. Results：The m RNA and protein expressions of PCDH10 were significantly up regulated by pc DNA3.1（＋）-PCDH10.Epirubicin could inhibit cell proliferation in a dose-ependent manner but the IC50 for PCDH10-transfected cells（0.87±0.07）μg/ml was much lower than that for vector-transfected cells（1.27±0.02）μg/ml（F=77.93,P=0.000）. The effect that epirubicin could induce cell escape could be inhibited by PCDH10（F=34.943,P=0.000）. The expression of CXCR4 which could be stimulated by epirubicin was decreased by PCDH10（F=117.269,P=0.000）. PCDH10 could also reduce the expression of Rho A（F=1 818.250,P=0.000）and MMP-9（F=523.764,P=0.000）obviously. Conclusion：PCDH10 could sensitize RPMI8226 cells to epirubicin and inhibit epirubicininduced cell escape. The down-regulating of CXCR4,Rho A and MMP-9 may act as an important mechanism.