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T-钙黏蛋白基因逆转黑素瘤细胞顺铂耐药性的体外研究

Effects of T-cadherin on Reversing Drug-resistance of Cisplatin to Melanoma in Vitro

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【作者】 陆海涛赵运华李雪飞于淑萍刘利君段昕所

【Author】 LU Haitao;ZHAO Yunhua;LI Xuefei;YU Shuping;LIU Lijun;DUAN Xinsuo;Department of Dermatology,Affiliated Hospital of Chengde Medical College;Chengde Central Hospital;

【机构】 承德医学院附属医院皮肤科承德市中心医院

【摘要】 目的探讨T-钙黏蛋白(T-cadherin)基因对黑素瘤顺铂(cisplatin,CDDP)耐药细胞株的逆转作用。方法采用大剂量冲击和逐步增加剂量相结合的方法诱导B16F10顺铂耐药细胞株(CDDP-R B16F10)。将T-cadherin转染肿瘤细胞。分别采用RT-PCR和Western blot法检测转染后T-cadherin mRNA和蛋白的表达。将T-cadherin与CDDP联合作用于CDDP-R B16F10细胞,采用MTT法、流式细胞术和Transwell侵袭实验检测细胞的增殖、早期凋亡和侵袭力。结果 RT-PCR和Western blot法检测表明,细胞可转录和表达T-cadherin。MTT法显示,T-cadherin联合CDDP可抑制CDDP-R B16F10细胞的增殖(P<0.05)。T-cadherin与CDDP联合对CDDP-R B16F10增殖的抑制有交互作用(P<0.05)。流式细胞术显示,T-cadherin联合CDDP可促进CDDP-R B16F10细胞的早期凋亡(P<0.05)。T-cadherin与CDDP联合对CDDP-R B16F10早期凋亡的促进有交互作用(P<0.05)。transwell侵袭实验显示,T-cadherin联合CDDP可抑制CDDP-R B16F10细胞的侵袭能力(P<0.05)。T-cadherin与CDDP联合对CDDP-R B16F10侵袭能力的抑制有交互作用(P<0.05)。结论 T-cadherin可逆转CDDP-R B16F10对CDDP的耐药性。

【Abstract】 Objective To investigate the reversal effect of T-cadherin gene on cisplatin resistance of melanoma cells. Methods CDDP resistance B16 F10(CDDP-R B16 F10) was induced by exposure to high and gradually increased dose of CDDP.The T-cadherin was transfected into B16 F10 cells. The expression of T-cadherin mRNA and protein were measured by RT-PCR and Western blot. The effects of T-cadherin combined with cisplatin on proliferation,early apoptosis and invasion of CDDP-R B16 F10 were determined by MTT assay,flow cytometry and Transwell invasion assay. Results The mRNA and protein of T-cadherin could be detected in T-cadherin-transfected cells. MTT assay showed that T-cadherin combined with CDDP could inhibit the proliferation of CDDP-R B16 F10 cells(P<0.05). There was interaction between T-cadherin and cisplatin in inhibiting the proliferation of CDDP-R B16 F10(P<0.05). Flow cytometry showed that T-cadherin combined with CDDP could induce early apoptosis of CDDP-R B16 F10 cells(P<0.05). There was interaction between T-cadherin and cisplatin in inducing early apoptosis of CDDP-R B16 F10(P<0.05). Transwell invasion assay showed that T-cadherin combined with CDDP could inhibit the invasive ability of CDDP-R B16 F10 cells(P<0.05). There was interaction between T-cadherin and cisplatin in inhibiting the invasiveness of CDDP-R B16 F10(P<0.05). Conclusion T-cadherin can reverse the cisplatin resistance of CDDP-R B16 F10.

  • 【文献出处】 中国皮肤性病学杂志 ,The Chinese Journal of Dermatovenereology , 编辑部邮箱 ,2019年01期
  • 【分类号】R734.2
  • 【被引频次】2
  • 【下载频次】79
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