天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (5): 459-463.doi: 10.11958/20181641

• 细胞与分子生物学 • 上一篇    下一篇

T-钙黏蛋白联合顺铂对黑色素瘤顺铂 耐药株的作用研究

陆海涛△，李雪飞，刘利君，何磊，段昕所   

  1. 作者单位：承德医学院附属医院皮肤性病科（邮编067000） 作者简介：陆海涛（1984），男，硕士研究生，主治医师，主要从事变态反应性皮肤病及皮肤良恶性肿瘤的诊治研究 △通讯作者 E-mail: PFKLHT@163.com
  • 收稿日期:2018-10-30 修回日期:2019-02-27 出版日期:2019-05-15 发布日期:2019-05-15
  • 通讯作者: 陆海涛 E-mail:PFKLHT@163.com

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

LU Hai-tao△, LI Xue-fei, LIU Li-jun, HE Lei, DUAN Xin-suo   

  1. Department of Dermatology, the Affiliated Hospital of Chengde Medical College, Chengde 067000, China △Corresponding Author E-mail: PFKLHT@163.com
  • Received:2018-10-30 Revised:2019-02-27 Published:2019-05-15 Online:2019-05-15

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摘要： 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-R B16F10）。MTT法检测CDDP-R B16F10 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（RT-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-R B16F10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-R B16F10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-R B16F10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。

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

Abstract: Abstract: 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.

Key words: cisplatin, melanoma, experimental, cell movement, T-cadherin, drug resistance；invasiveness