穿膜融合多肽TAT-N24对结肠癌细胞增殖的影响

Cell-permeable TAT-N24 Fusion Peptide Inhibiting Proliferation of Colon Cancer Cells

目的观察穿膜融合多肽TAT-24对结肠癌细胞增殖的影响。方法观察TAT-N24对HT29细胞内蛋白表达的影响,流式细胞仪检测HT29细胞周期进程,应用BrdU掺入的方法检测TAT-N24对细胞DNA合成的影响,证实TAT-N24的抗肿瘤作用。结果 Western blot结果显示TAT-N24能够有效进入细胞内,TAT-N24处理的细胞内Rb蛋白的表达无显著变化,但可见磷酸化Rb蛋白表达量显著降低。BrdU/PI双掺入法检测细胞DNA合成结果显示,对照组BrdU阳性细胞数占总细胞的比例为(52.2±1.88)%,而TAT-N24组BrdU阳性细胞数占总细胞的比例为(29.9±2.14)%,两组间比较差异有统计学意义(P<0.05)。对照组HT29细胞中G0/G1期细胞为(55.27±2.48)%,S期和G2/M期细胞数分别为(26.97±0.94)%和(17.76±1.77)%,而TAT-N24组HT29细胞G0/G1期细胞减少到(65.10±1.79)%,S期和G2/M期细胞分别为(18.49±0.68)%和(16.40±1.51)%,较对照组差异有统计学意义(P<0.05)。结论融合多肽TAT-24能有效抑制Rb蛋白磷酸化,阻滞结肠癌HT29细胞周期进程,抑制细胞DNA合成。

Objective To elucidate the role of cell-permeable TAT-N24 fusion peptide inhibiting proliferation of colon cancer cells.Methods Detect the cell cycle progression by flow cytometer,analyze the DNA synthesis by BrdU/PI method and certify the roles of TAT-N24 in the tumor.Results Western blot shows that TAT-N24 can enter HT29 cells. With the treatment of TAT-N24,the Rb phosphorylation is decreased.In the control group,the number of BrdU positive cells is （52.2± 1.88 ）%,while the TAT-N24 group is （29.9±2.14）%.In control group,cell cycle analysis shows that the number of cells in G0/G1 phases is（55.27±2.48）%,in S and G2/M phases cells are （26.97± 0.94 ）% and （17.76±1.77）%,but in TAT-N24 group,the number of cells in G0/G1 phases is （65.10 ±1.79）%,the S and G2/M phases cells are （18.49± 0.68 ）% and （16.40±1.51）%.Conclusion TAT-N24 fusion peptide can inhibit the Rb protein phosphorylation,induce the cell cycle arrest and inhibit the DNA synthesis in colon cancer cells.