摘要
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 the invasiveness of malignant melanoma. Methods B16F10 cells were cultured in vitro. Blank vector pEGFP-N1 and pEGFP-N1-T-cadherin were used to transfeet the B16F10 cells. The expression of T-cadherin mRNA and protein were measured by reverse transcription polymerase chain reaction （RT-PCR） and SP immunohistochemistry method. Cell invasiveness was determined by Matrigel Transwell in vitro invasion assay. Results The mRNA and protein of T-cadherin could be detected in T-cadherin-transfected cells; The number of invasion cells was significantly lower in T-cadherin-transfected cells （2.20 ± 0.49） compared with the un-transfected cells （16.80± 1.39）and blank vector transfectants （15.00± 1.26）,with a significant difference（ P 〈 0.05）. There were no statistical difference between untransfected cells and blank vector transfectauts（P 〉 0.05 ） . MTT assay showed that the OD value of invasion cells was obviously lower in T-cadherin-transfected cells （0. 124 ± 0. 003 ） than that of un-transfected cells （0.263 ± 0.006） and blank vector transfectants （0.247± 0.012） , with a significant difference （ P 〈 0.05 ）. There were no statistical difference between un-transfected cells and blank vector transfectants （P 〉 0.05 ）. Conclusion T-cadherin suppresses the invasive ability of malignant melanoma cells in vitro.
出处
《中国皮肤性病学杂志》
CAS
北大核心
2011年第5期329-332,共4页
The Chinese Journal of Dermatovenereology
基金
河北省科技支撑计划项目
