T-钙黏蛋白对小鼠皮下氮烯咪胺耐药黑素瘤B16F10细胞株的影响

The Effects of T-cadherin on Dacarbazine-resistant Subcutaneous B16F10 Melanoma in Mice

目的观察T-钙黏蛋白(cadherin)对小鼠皮下氮烯咪胺(dacarbazine,DTIC)耐药黑素瘤B16F10细胞株的影响。方法通过体外分步诱导法诱导B16F10氮烯咪胺耐药细胞株(DTIC-R B16F10)。采用CCK-8法检测DTIC-R B16F10细胞的增殖情况。将T-cadherin基因转染至DTIC-R B16F10细胞。免疫组织化学检测转染后T-cadherin的表达情况。培养DTIC-R B16F10细胞,分为6组:对照组、pEGFP-N1组、T-cadherin组、DTIC组、pEGFP-N1联合DTIC组和T-cadherin联合DTIC组。通过CCK-8法和Transwell小室实验检测T-cadherin与氮烯咪胺联合对DTIC-R B16F10细胞活性和迁移的影响。建立荷瘤小鼠模型,观察T-cadherin对小鼠皮下注射DTIC-R B16F10细胞后肿瘤生长的影响。采用析因分析法评价T-cadherin联合氮烯咪胺对细胞活性、迁移和小鼠皮下瘤生长的影响。结果DTIC-R B16F10组、B16F10组与DTIC-R B16F10+DTIC组A值差异无统计学意义(P>0.05)。免疫组织化学检测表明细胞表达T-cadherin。T-cadherin联合DTIC组细胞活性、迁移细胞数及肿瘤重量均显著低于其他组(P<0.05)。析因分析显示,T-cadherin联合氮烯咪胺对DTIC-R B16F10的细胞活性、迁移和小鼠皮下瘤生长的抑制有协同作用。结论T-cadherin可抑制黑素瘤小鼠皮下瘤的生长,增加黑素瘤对氮烯咪胺的敏感性。

Objective To observe the effects of T-cadherin on dacarbazine-resistant subcutaneous B16 F10 melanoma in mice.Methods Dacarbazine(DTIC)resistance in B16 F10(DTIC-R B16 F10)was achieved by in vitro stepwise induction.Cell counting kit-8(CCK-8)was used to test the proliferation of DTIC-R B16 F10 cells.T-cadherin gene was transfected into DTIC-R B16 F10 cells.The expression of T-cadherin was measured by immunohistochemistry.DTIC-R B16 F10 cells were cultured and divided into six groups:control group,pEGFP-N1 group,T-cadherin group,DTIC group,pEGFP-N1 combined with DTIC group and T-cadherin combined with DTIC group.The effects of T-cadherin combined with DTIC on cell viability and migration of DTIC-R B16 F10 were determined by CCK8 and transwell assay,respectively.Tumor-bearing mice were established.The effect of T-cadherin on the kinetics of tumor growth after subcutaneous injection of DTIC-R B16 F10 cells in mice was observed.Factorial analysis was used to evaluate the effects of T-cadherin combined with DTIC on cell viability,migration and growth of subcutaneous tumors in mice.Results There were no statistical differences in A value among DTIC-R B16 F10 group,B16 F10 group and DTIC-R B16 F10+DTIC group.The expression of T-cadherin was measured by immunohistochemistry.The cell viability,cell migration and tumor weight were significantly lower in T-cadherin combined with DTIC group than in other groups(P<0.05).The factorial analysis showed that there were synergistic effects of T-cadherin and DTIC in inhibiting the cell viability,migration and growth of subcutaneous tumors in mice(P<0.05).Conclusion T-cadherin shows a significant inhibitory effect on the growth of subcutaneous melanoma in mice.T-cadherin can promote the sensitivity of melanoma to DTIC.