T-钙黏蛋白对B16F10黑素瘤细胞周期调节的研究

The research of T-cadherin on the cell cycle of B16F10 melanoma cell

目的探讨T-钙黏蛋白对B16F10黑素瘤细胞周期的影响。方法应用脂质体LipofectamineTM2000将构建的pEGFP-N1-T-钙黏蛋白以及pEGFP-N1空载体转染至B16F10细胞,OPTI-MEM培养基体外培养6 h后RPMI-1640继续培养24 h,应用碘化丙啶标记,流式细胞仪检测T-钙黏蛋白对B16F10黑素瘤细胞周期的影响。结果转染T-钙黏蛋白组G0/G1期和G2/M期细胞比率分别为(60.917±1.897)%、(8.513±0.651)%,明显高于未转染组的(53.563±1.856)%、(2.627±0.434)%和pEGFP-N1空载体组的(53.880±2.164)%、(2.770±0.466)%,差异有显著性(P<0.05);转染T-钙黏蛋白组S期细胞比率为(30.570±1.368)%,低于未转染组的(43.810±2.003)%及pEGFP-N1空载体组的(43.350±2.629)%,差异有统计学意义(P<0.05)。未转染组与转染pEGFP-N1空载体组各期差异均无统计学意义(P>0.05)。结论 T-钙黏蛋白使B16F10细胞G0/G1期和G2/M期比率增高,S期比率降低。

Objective To investigate the effect of T - cadherin on the cell cycle of B16FIO melanoma cell. Methods pEGFP - N1 - T - cadherin and Blank vector pEGFP - N1 were used to transfect the B16F10 cells with LipofectamineTM2000. These cells were cultured with RPMI - 1640 24 hour after using OPTI - MEM 6 hour in vitro. The cell cycle of B16FIO melanoma were measured by flow cytometry. Results The percentage of cells in G0/G1 and G2/M phase were higher in T- cadherintransfected ceils （60. 917 ± 1. 897 ）% and （8.513 ± 0. 651 ）% compared with the （53. 563 ±1. 856） % and （ 2. 627 ± 0. 434） % of un - transfected ceils, （53. 880 ± 2. 164 ） % and （2. 770 ±0. 466 ） % in blank vector transfectants. There was significant difference（ P 〈0.05）. The percentage of cells at S phase was lower in T - cadherin - transfected cells with （30. 570 ± 1. 368 ） % compared with the un - transfected cells with （43.810 ＋ 2. 003 ） % and blank vector transfectants with （ 43. 350 ± 2. 629 ） %. There was significant difference （ P 〈 0. 05 ）. There were no statistical difference between blank vector transfectants and un - transfected cells （ P 〉0.05）. Conclusion T - cadherin can make the percentage of B16F10 melanoma cells in G0 / G1 and G2 / M phase high than other two groups , low in S stage.