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目的探讨苯乙酸(PA)对结直肠癌HCT-8细胞株细胞增殖及同源盒(HOX)基因表达的影响。方法应用噻唑蓝(MTT)比色法,以1.0、2.0、3.0、4.0、5.0 mmol/L的PA作用于HCT-8细胞,分别于244、8、72 h对细胞增殖进行检测。根据引物的不同,将已知的22种HOX基因分为3组(P1、P2、P3),应用半定量逆转录-聚合酶链反应(RT-PCR)检测应用PA前后3组HOX基因mRNA的表达水平。HOX基因(组)表达水平用基因(组)与β肌动蛋白(-βactin)灰度比值表示。结果1.0~5.0 mmol/L PA作用HCT-8细胞24~72 h,随着PA浓度的增加或作用时间的延长,细胞生长抑制率明显增加,PA作用24 h为5.1%~24.3%,48 h为16.7%~72.3%,72 h为30.2%~93.4%。RT-PCR法检测,结直肠癌HCT-8细胞中P1组HOX基因表达(0.770 1±0.088 3)明显强于P2组(0.122 1±0.078 2)、P3组(0.180 6±0.081 1,P<0.01)。应用2.0 mmol/L PA作用72 h后,P1组HOX基因表达(0.578 1±0.083 6)显著减弱,P2、P3组HOX基因表达(0.394 1±0.081 9)(、0.560 1±0.073 6)显著增强,与用药前比较,差异有统计学意义(P<0.01)。结论PA可有效抑制结直肠癌HCT-8细胞的增殖;PA对HOX基因具有明显的调控作用,HOX基因有望成为结直肠癌基因治疗靶点。
Objective To investigate the effects of phenylacetic acid (PA) on cell proliferation and homeobox (HOX) gene expression in colorectal cancer HCT-8 cell line. Methods HCT-8 cells were treated with 1.0, 2.0, 3.0, 4.0 and 5.0 mmol / L PA by MTT assay. The cell proliferation was detected at 244, 8 and 72 hours respectively. According to the different primers, 22 known HOX genes were divided into three groups (P1, P2 and P3). HOX mRNA was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) The expression level. The expression level of HOX gene (group) is expressed as the ratio of gray scale of gene (group) and β-actin (-βactin). RESULTS: HCT-8 cells were treated with 1.0-5.0 mmol / L PA for 24-72 h. With the increase of concentration of PA or the prolongation of action time, the growth inhibition rate of HCT-8 cells was significantly increased. The effect of PA for 24 h was 5.1% -24.3% For 16.7% -72.3%, 72.2% for 30.2% -93.4%. The expression of HOX gene in P1 group was significantly higher than that in P2 group (0.122 1 ± 0.078 2), P3 group (0.180 6 ± 0.081 1, P < 0.01). HOX gene expression in P1 group was significantly decreased (0.578 1 ± 0.083 6) after treated with 2.0 mmol / L PA for 72 h, but HOX gene expression in group P2 and P3 was significantly increased (0.394 ± 0.081 9, 0.560 1 ± 0.073 6) , Compared with before treatment, the difference was statistically significant (P <0.01). Conclusion PA can effectively inhibit the proliferation of colorectal cancer HCT-8 cells; PA has obvious regulation on HOX gene, HOX gene is expected to become a target of colorectal cancer gene therapy.