曲古菌素A对人胰腺癌PaTu-8988细胞增殖和细胞周期的影响

目的 观察曲古菌素A(tfichostatin A,TSA)对人胰腺癌PaTu-8988细胞增殖及细胞周期的影响.方法 不同浓度(0.1、0.5、2.0 μmol/L)TSA处理人胰腺癌PaTu-8988细胞,并设空白对照组.采用WST-8法检测细胞存活率;流式细胞仪检测细胞周期的变化;实时定量PCR检测细胞周期相关基因p21和cyclin DI mRNA的表达.结果 对照组、TSA 0.1μmol/L、0.5μmol/L和2.0μmol/L组的细胞存活率分别为(100.0±4.2)%、(88.5±4.2)%、(79.7±5.0)%和(64.3±7.2)%,各TSA组均显著低于对照组(P<0.01).TSA 0.1μmoL/L、0.5μmol/L组的细胞以G1期居多,2.0μmol/L组(50.29±7.53)%细胞阻滞在G2/M期.TSA各组p21 mRNA表达值分别为5.29±1.16、7.79±0.41、8.61±0.73,较对照组l,00±0.08显著升高(P<0.01);cyclin DI mRNA表达值分别为1.13±0.12、0.42±0.06、0.19±0.06,较对照组1.00±0.07表达降低(P<0.05).结论 TSA通过上调p21及下调cyclin D1基因的表达,导致细胞周期阻滞.

Effects of inhibitors of trichostatin A on the growth and cell cycle of PaTu-8988 cell lines

Objective To investigate the effect of trichostatin A (TSA) on inhibition of cell proliferation and cell cycle of human pancreatic cancer cell line PaTu-8988 in vitro. Methods PaTu-8988 cells were treated with different concentration TSA (0.1,0.5,2.0μmol/L), and blank control group was used. The growth inhibition rates of cells were measured by WST-8 method. Cell cycle and apoptosis were detected by flow cytometry; the expressions of p21 mRNA and HDAC1 mRNA were measured by Real-Time PCR. Results Survival rates of TSA 0.1μmol/L,0.5μmol/Land2.0μmol/L group was (88.5±4.2)%, (79.7±5.O)% and (64.3±7.2)%, respectively, which were all significantly lower than (100.0±4.2)% of the blank control group (P<0.01). Cells in the group of TSA O.1μmol/L and 0.5μmol/L were hindered at G1 phase, while (50.29±7.53)% of the cells in the group of TSA 2.0μmol/L were hindered at G2/M phase. The expression rates of p21 mRNA in different TSA group were 5.29±1.16, 7.79±0.41, 8.61± 0.73, respectively, which were higher than that in the control group (1.00±0.08, P<0.01); the expression rates of Cyclin DI mRNA were 1.13±0.12, 0.42±0.06, 0.19±0.06, respectively, which were higher than that in the control group (1.00±0.07, P<0.05). Conclusions TSA may up-regulate the expression of p21 and down-regulate the expression of cyclin D1, and induce cell cycle blockade.