摘要
目的：观察三氧化二砷（arsenic trioxide，As，O，）对人膀胱癌T24细胞增殖及凋亡蛋白酶活化因子一1（apoptotic protease activating factor-1，Ap．f-，）基因表达的影响。方法：取处于对数生长期的T24细胞，用0．25％胰酶消化成细胞密度为1×10^5／mL的单细胞悬液，再用含有不同浓度（0、1、2、4、8μmol／L）As2O，的培养基作为药物组，并设置空白对照组，置于培养箱中分别培养24、48和72h后，用二甲基四氮唑蓝（M1v11法检~lJAs，O，对T24细胞的增殖抑制率；取T242~I胞在不同浓度（0、1、2、4、8μmot／L）的As，O，培养液中培养72h后用脱氧核糖核酸原位末端转移酶标记技术（TUNEL）法检测细胞凋亡，用RT—PCR及Westernblouing检测As2O，对Apaf-1mRNA和蛋白表达的影响。结果：与对照组相比，2、4、8μmol／LAs2O，剂量组能有效抑制T24细胞增殖，且具有剂量一反应关系（24h时iv=-0．962，P=0．038；48h时r=0．959，P=0．041；72h时r=-0．973，P=0．027）。As，O，于1～8μmol／L作用72h时细胞出现典型的凋亡变化，具有剂量-反应关系（r=0．993，P=0．007）；同时Apaf-1mRNA和蛋白的表达均明显增强，且具有剂量一反应关系（mRNA：r=0．986，P=O．014；蛋白：r=0．989，P=O．ooo）。结论：As2O3能够有效抑制膀胱癌T24细胞生长、增殖并诱导其凋亡，机制可能与其Y~NApaf-，基因的表达有关。
OBJECTIVE： To observe the effects of arsenic trioxide （As203） on human bladder cancer T24 cell proliferation and apoptosis protease activating factor-1 （Apaf-1） gene expression. METHODS： T24 cells in the logarithmic growth phase, were digested with 0.25% trypsin into a cell density of 1 ~ 105/mL single cell suspension. Then cells were inculated with different concentrations （0, 1, 2, 4, 8 tx mol/L） of As203, as well as blank control group for 24, 48 and 72 h. Cell proliferation inhibition rate was evaluated by MTT. After cell treatment with As203 （0, 1, 2, 4, 8 Ix mol/L） for 72 h, deoxyribose situ terminal transferase labeling （TUNEL） method was used to measure apoptosis ; RT- PCR and Western blotting to assess the impact of As203 on Apaf-1 mRNA and protein expression. RESULTS ： Compared with the control group, the 2, 4, 8 ~ mol/L As203 dose groups could effectively inhibit the proliferation of T24 cells in a concentration-dependent manner （24 h, r=0.962, P=0.038; 48 h, r=0.959, P=0.041; 72 h, r=0.973, P=0.027）. Cells treated with As203 at 1-8 tx mol/L for 72 h, showed typical apoptotic changes in a concentration-dependent manner （r= 0.993, P=0.007）; whilst Apaf-1 mRNA and protein expressions were also significantly increased in a concentration- dependent（mRNA, r=0.986, P=0.014; protein, r=0.989, P=0.000）. CONCLUSION： As203 could effectively inhibit bladder cancer T24 cell growth, proliferation and induce apoptosis. The possible mechanism may be related to increased Apaf-1 gene expression.
出处
《癌变·畸变·突变》
CAS
CSCD
2013年第5期338-342,共5页
Carcinogenesis,Teratogenesis & Mutagenesis
关键词
三氧化二砷
APAF-1
细胞凋亡
膀胱肿瘤
arsenic trioxide
apoptotic protease activating factor-1
apoptosis
bladder tumor
