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目的探讨T-钙黏蛋白(T-cadherin)对黑素瘤侵袭力的影响。方法体外培养黑素瘤细胞株B16F10,将pEGFP-N1空载体和构建的pEGFP-N1-T-cadherin转染B16F10细胞,采用RT-PCR和免疫组化SP法检测T-cadherin基因及蛋白水平的表达情况;应用transwell细胞侵袭实验测定转染T-cadherin后肿瘤细胞的侵袭力。结果 RT-PCR和免疫组化SP法检测结果表明,细胞可转录和表达T-cadherin;转染T-cadherin组穿过transwell小室膜的细胞数目(2.20±0.49)明显少于未转染组(16.80±1.39)和转染pEGFP-N1空载体组(15.00±1.26),差异有统计学意义(P<0.05),未转染组与转染pEGPF-N1空载体组差异无统计学意义(P>0.05);MTT法结果显示,转染T-cadherin组穿过小室膜细胞的吸光度(OD)值(0.124±0.003)明显低于未转染组(0.263±0.006)和转染pEGFP-N1空载体组(0.247±0.012),差异有统计学意义(P<0.05),未转染组与转染pEGFP-N1空载体组的差异无统计学意义(P>0.05)。结论 T-cadherin对黑素瘤细胞的体外侵袭能力有抑制作用。
Objective To investigate the effect of T-cadherin on invasiveness of melanoma. Methods Melanoma cell line B16F10 was cultured in vitro. The empty vector pEGFP-N1 and the constructed pEGFP-N1-T-cadherin were transfected into B16F10 cells. The expression of T-cadherin gene and protein was detected by RT-PCR and immunohistochemistry The transwell cell invasion assay was used to determine the invasiveness of tumor cells after T-cadherin transfection. Results The results of RT-PCR and immunohistochemistry showed that the number of cells transfected with T-cadherin in T-cadherin group was significantly lower than that in untransfected group (2.20 ± 0.49) 16.80 ± 1.39) and pEGFP-N1 empty vector group (15.00 ± 1.26) respectively, the difference was statistically significant (P <0.05) > 0.05). The results of MTT assay showed that the OD value of transfected T-cadherin group (0.124 ± 0.003) was significantly lower than that of untransfected group (0.263 ± 0.006) and transfected pEGFP-N1 null (0.247 ± 0.012), the difference was statistically significant (P <0.05). There was no significant difference between untransfected group and pEGFP-N1 empty vector group (P> 0.05). Conclusion T-cadherin can inhibit the invasion of melanoma cells in vitro.