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基因Livin转染对膀胱癌细胞凋亡的影响

[Effects of gene Livin transfection affect on the apoptosis in bladder carcinoma cells].

作者信息

Liu Xian-Kui, Liu Hai-Bo, Kong Chui-Ze

机构信息

Department of Urology, First Affiliated Hospital of China Medical University, Shenyang 110001, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2008 Mar 25;88(12):853-5.

Abstract

OBJECTIVE

To investigate the effect of gene Livin transfection on the apoptosis of human bladder transitional cell carcinoma (BTCC) cells.

METHODS

Target fragment containing Livin full-length cDNA was obtained from the breast cancer cells of the line MCF7. Vector pcDNA3. 1 (+)-Livin was constructed and transfected into the bladder transitional cell carcinoma (BTCC) cells of the line T24 [T24/ Livin+ cells]. Another T24 cells were transfected with the eukaryotic expression vectors pcDNA3.1 (+) [T24/pcDNA3.1 (+) cells]. T24 cells without transfection were used as blank control group. These T24 cells underwent G418 selection so as to screen the T24/Livin+ cells stably expressing the target fragment. RT-PCR was used to detect the Livin expression level in the transfected cells. After the cells were treated with mitomycin C (MMC) for 24 h, MTT method was used to detect the inhibition rate. Acridine orange (AO) staining was used to observe the apoptotic cells. The apoptotic rate was observed by flow cytometry.

RESULTS

RT-PCR results indicated that expression of Livin was positive in the T24/Livin+ cells, while were negative in the T24/pcDNA3.1 (+) and T24 cells. After being treated with MMC for 24 h, the apoptotic rate of the T24/Livin+ cell was (8.7 +/- 1.5)%, significantly lower than those of the T24/pcDNA3.1 (+) cells and T24 cells [(21.4 +/- 2.3)% and (19.6 +/- 2.3)% respectively, both P < 0.01].

CONCLUSION

The gene Livin increases the anti-apoptosis ability of tumor cells and the cell apoptosis induced by chemotherapy.

摘要

目的

探讨Livin基因转染对人膀胱移行细胞癌(BTCC)细胞凋亡的影响。

方法

从MCF7乳腺癌细胞系中获取含Livin全长cDNA的目的片段,构建载体pcDNA3.1(+)-Livin并转染至T24膀胱移行细胞癌(BTCC)细胞系中[T24/Livin+细胞]。另将真核表达载体pcDNA3.1(+)转染至T24细胞中[T24/pcDNA3.1(+)细胞]。未转染的T24细胞作为空白对照组。对这些T24细胞进行G418筛选,以筛选出稳定表达目的片段的T24/Livin+细胞。采用RT-PCR检测转染细胞中Livin的表达水平。细胞经丝裂霉素C(MMC)处理24 h后,采用MTT法检测抑制率。采用吖啶橙(AO)染色观察凋亡细胞。通过流式细胞术观察凋亡率。

结果

RT-PCR结果显示,Livin在T24/Livin+细胞中表达呈阳性,而在T24/pcDNA3.1(+)细胞和T24细胞中呈阴性。经MMC处理24 h后,T24/Livin+细胞的凋亡率为(8.7±1.5)%,显著低于T24/pcDNA3.1(+)细胞和T24细胞[分别为(21.4±2.3)%和(19.6±2.3)%,P均<0.01]。

结论

Livin基因增强了肿瘤细胞的抗凋亡能力以及化疗诱导的细胞凋亡。

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