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目的探讨β-胡萝卜素抑制人膀胱癌T24细胞的分子生物学机制。方法不同浓度的β-胡萝卜素作用于T24细胞,利用MTT法检测细胞增殖,流式细胞术检测其凋亡指数,半定量RT-PCR检测Cx43、Bcl-2、Bax mRNA的变化,Western blot检测Cx43、Bcl-2、Bax蛋白的表达。结果 10和20μmol/L组β-胡萝卜素均能明显抑制T24细胞的生长和诱导细胞凋亡,其抑制率分别为28.83%、63.02%,并且呈明显的剂量-效应关系(P<0.05),凋亡率分别为0.126±0.022和0.190±0.024(P≤0.01),随β-胡萝卜素浓度增加,细胞凋亡率增大,呈剂量依赖性关系(P=0.002)。与对照组相比,10和20μmol/L组β-胡萝卜素上调Cx43 mRNA及其蛋白的表达,增强细胞间的细胞间隙连接通讯(GJIC)功能(P<0.05),并且呈剂量依赖关系(P≤0.01);下调Bcl-2 mRNA及其蛋白的表达,并且呈剂量依赖关系(P≤0.01);Bax mRNA及其蛋白的表达有上调趋势,但与对照组比较,差异无显著性。结论β-胡萝卜素(10,20μmol/L)能通过有效下调Bcl-2 mRNA及其蛋白在T24细胞的表达,上调T24细胞Cx43转录水平的表达、增强细胞间隙连接所介导的GJIC功能,从而诱导膀胱肿瘤细胞凋亡,抑制其生长。
Objective To investigate the molecular mechanism of β-carotene inhibiting human bladder cancer T24 cells. Methods Different concentrations of β-carotene were applied to T24 cells. MTT assay was used to detect cell proliferation. Flow cytometry was used to detect apoptosis index. Cx43, Bcl-2 and Bax mRNA were detected by semi-quantitative RT-PCR. Cx43, Bcl-2, Bax protein expression. Results Both β-carotene at 10 and 20 μmol / L significantly inhibited the growth of T24 cells and induced apoptosis. The inhibitory rates were 28.83% and 63.02%, respectively, with significant dose-effect relationship (P <0.05) The apoptotic rates were 0.126 ± 0.022 and 0.190 ± 0.024, respectively (P ≤ 0.01). With the increase of β-carotene concentration, the apoptosis rate increased in a dose-dependent manner (P = 0.002). Compared with the control group, β-carotene at 10 and 20 μmol / L upregulated the expression of Cx43 mRNA and protein, increased the cell-cell gap junctional intercellular communication (GJIC) function in a dose-dependent manner ≤0.01). The expression of Bcl-2 mRNA and its protein was down-regulated in a dose-dependent manner (P≤0.01). The expression of Bax mRNA and its protein were up-regulated. However, there was no significant difference compared with the control group. Conclusion β-carotene (10 and 20 μmol / L) can up-regulate the expression of Bx-2 mRNA and its protein in T24 cells and up-regulate the expression of Cx43 in T24 cells, and enhance the GJIC function mediated by gap junctions Induce bladder cancer cell apoptosis and inhibit its growth.