T-钙黏蛋白联合顺铂对黑色素瘤顺铂耐药株的作用研究  

The effect of T-cadherin combined with cisplatin on cisplatin resistant malignant melanoma cell line

作　　者：陆海涛[1] 李雪飞[1] 刘利君[1] 何磊[1] 段昕所[1] LU Hai-tao;LI Xue-fei;LIU Li-jun;HE Lei;DUAN Xin-suo(Department of Dermatology,the Affiliated Hospital of Chengde Medical College,Chengde 067000,China)

机构地区：[1]承德医学院附属医院皮肤性病科

出　　处：《天津医药》2019年第5期459-463,I0001,共6页Tianjin Medical Journal

摘　　要：目的探讨T-钙黏蛋白联合顺铂对黑色素瘤顺铂耐药株的作用。方法采用大剂量冲击和逐步增加剂量相结合的方法诱导建立小鼠黑色素瘤B16F10顺铂耐药细胞株(CDDP-RB16F10)。 MTT法检测CDDP-RB16F10的增殖能力。将T-钙黏蛋白转染肿瘤细胞。分别采用反转录聚合酶链式反应(R T-PCR)和免疫组化SP法检测转染后T-钙黏蛋白mRNA和蛋白的表达。实验将CDDP-RB16F10细胞分为空白对照组、pEGFP-N1组、pEGFP-N1-T-cadherin组、顺铂组、pEGFP-N1联合顺铂组、pEGFP-N1-T-cadherin联合顺铂组。采用Wound-healing划痕实验和transwell侵袭实验检测T-钙黏蛋白联合顺铂对CDDP-RB16F10细胞迁移和侵袭力的影响。结果成功建立黑色素瘤顺铂耐药细胞株。MTT法结果显示,CDDP-RB16F10细胞增殖能力与B16F10细胞相比差异无统计学意义(P>0.05)。 RT-PCR和免疫组化SP法检测表明,转染后细胞可稳定转录和表达T-钙黏蛋白。pEGFP-N1-T-cadherin联合顺铂组细胞迁移率和穿膜细胞数低于pEGFP-N1-T-cadherin组(P<0 .05),而 pEGFP-N1-T-cadherin组低于空白对照组、pEGFP-N1组、顺铂组和pEGFP-N1联合顺铂组(P<0 .05)。析因分析显示,T-钙黏蛋白与顺铂联合对CDDP-R B16F10细胞迁移率及侵袭力的抑制有交互作用(P<0 .05)。结论 T-钙黏蛋白可恢复顺铂对黑色素瘤顺铂耐药细胞株迁移及侵袭力的抑制作用。Objective To investigate the effect of T-cadherin combined with cisplatin on cisplatin resistant malignant melanoma cell line. Methods CDDP resistance B16F10 (CDDP-R B16F10) was induced by using high and gradually increased dose of CDDP.MTT assay was used to test the proliferation of CDDP-R B16F10. The T-cadherin was transfected into CDDP-R B16F10 cells. The expressions of T-cadherin mRNA and protein were measured by reverse transcription polymerase chain reaction (RT-PCR) and SP immunohistochemistry method. There were six groups in this study including control group, pEGFP-N1 group, pEGFP-N1-T-cadherin group, cisplatin group, pEGFP-N1 combined with cisplatin group and pEGFP-N1-T-cadherin combined with cisplatin group. The effects of T-cadherin combined with cisplatin on migration and invasion of CDDP-R B16F10 were determined by Wound-healing assay and Transwell invasion assay. Factor analysis method was used to evaluate the interactions of T-cadherin and CDDP on migration and invasion of CDDP-R B16F10. Results The CDDP-R B16F10 cell line was successfully established. There was no statistical difference in proliferation between CDDP-R B16F10 cells and B16F10 cells (P>0.05). RT-PCR and SP immunohistochemistry assay showed that T- cadherin could be transcribed and expressed in cells. The cell migration rate and transmembrane number were significantly lower in pEGFP-N1-T-cadherin combined with cisplatin group than those of pEGFP-N1-T-cadherin group (P<0.05). The cell migration rate and transmembrane number were significantly lower in pEGFP-N1-T-cadherin group than those of control group, pEGFP-N1 group, cisplatin group and pEGFP-N1 combined with cisplatin group (P<0.05). There were interaction between T-cadherin and cisplatin in inhibiting the migration and invasiveness of CDDP-R B16F10 (P<0.05).Conclusion T-cadherin gene can restore the inhibitory effect of cisplatin on the migration and invasiveness of cisplatin resistant melanoma cell line.

关 键 词：顺铂 黑色素瘤 实验性 细胞运动 T-钙黏蛋白 耐药 侵袭力 

分 类 号：R739.5[医药卫生—肿瘤]