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姜黄素诱导人膀胱癌UMUC2细胞株凋亡的作用

Effects of Curcumin on Inducing Apoptosis in Human Bladder Cancer UMUC2 Cells

  • 摘要: 目的为寻找有效无毒的膀胱灌注药物用于膀胱肿瘤术后预防肿瘤种植与复发,我们观察了姜黄素对人膀胱癌细胞UMUC2的细胞毒效应及诱导凋亡的体外作用。方法以0~160 μmol/L姜黄素作用于UMUC2细胞1~48 h,应用MTT法、平板克隆实验观察姜黄素的细胞毒效应,荧光染色观察凋亡形态学改变,流式细胞仪进行细胞周期分析及凋亡定量测定,免疫细胞化学染色观察凋亡相关蛋白p53和Survivin的表达。结果所有浓度组姜黄素均对膀胱癌UMUC2细胞有明显的生长抑制作用,160 μmol/L姜黄素作用1 h对全部癌细胞是致命的。荧光染色观察到典型的核凝集、碎裂凋亡形态学改变。流式细胞术定量分析了亚G1峰,后者同时表明姜黄素导致的细胞周期阻滞以G2/M期为主,使S期细胞比例显著减少。免疫细胞化学染色显示姜黄素下调p53、Survivin蛋白的表达。结论姜黄素明显抑制膀胱癌细胞系UMUC2的生长并诱导凋亡,导致细胞G2/M期阻滞,其作用机制与其下调p53、Survivin的表达相关,克隆形成实验证实大剂量姜黄素、短期作用对膀胱癌细胞系UMUC2有致命的细胞毒效应,显示了姜黄素用于膀胱癌患者的化学治疗,尤其是腔内灌注化疗的可能性。

     

    Abstract: ObjectiveTo investigate the cytotoxic effects and the induction of apoptois of curcumin on purinary bladder UMUC2 cancer cell lines in vitro;and develop an effective nontoxic intravesical agent that may be used immediately after bladder tumor resection to prevent the implantation of tumor cells and recurrence. significant clinical advancement. Methods UMUC2 cells were incubated with 0 to 160 μmol/L curcumin for 1 to 48 h. The cell viability was determined by MTT and clonal assay.The morphology of apoptotic cells was assessed by fluorescence microscopy after acridine orange/ethidium bromide (AO/EB) staining.Quantification of apoptosis and detection of cell cycle redistribution were evaluated by flow cytometry.The expression of apoptosis-associated protein p53 and Survivin was detected by immunocytochemical staining. Results Curcumin with different concentration could significantly result in the cell growth suppression.At the 160 μmol/L dose of curcumin was completely lethal to the UMUC2 cell lines after 1h treatment on clonal growth assay.Fluorescence microscopy and FCM revealed that the condensation and fragmentation of their nuclei and sub-G1 region after curcumin treatment respectively.The later also revealed cell phase arrest induced by curcumin was G2/M phase arrest mainly with significant decrease of S phase.Immunocytochemical staining revealed that the anti-apoptotic p53 and Survivin protein was down-regulated by the curcumin treatment. Conclusion Curcumin can suppress the growth,induce apoptosis of bladder cancer UMUC2 cell in vitro.These results suggested that the inhibition of p53 and survivin expression in UMUC2 cells might account for the mechanism of curcumin-induced apoptosis.At the 160 μmol/L concentration of curcumin is a potent cytotoxic agent against the UMUC2 bladder tumor cell lines.Taken together,our present findings showed the promising anti-cancer efficacy of curcumin against human bladder cancer cells,especially,as an intravesical agent.

     

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