Tianjin Med J ›› 2019, Vol. 47 ›› Issue (5): 459-463.doi: 10.11958/20181641

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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

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