Radiation-inducible promoters mediated PUMA gene in the treatment of xenograft of human tongue squamous in naked mice

doi:10.3877/cma.j.issn.1674-1366.2012.05.006

Abstract

Abstract:

Objective

To evluate the therapeutic effect of PUMA gene mediated by radiationinducible promoters in the treatment of xenograft of human tongue squamous carcinoma in nude mice.

Methods

The recombinant pcDNA3.1 （+）/E-PUMA was constructed， and the model of transplanted Tca8113 cell line in naked mice was founded. The plasmids pcDNA（+）3.1/E-PUMA were transfected into tumors by lipofectamine. 24 h later， the tumors were treated by 3 Gy irradiations. Animals were monitored for volume of tumors at regular intervals. PUMA mRNA was detected by RT-PCR. PCNA and Apoptosis were detected by immuno-histiochemical method and in situ end-labeling respectively. The data was analyzed by SPSS 11.0 software package for chi-square test.

Results

The tumors were suppressed obviously by radiation-inducible promoters mediated PUMA gene. The 3 Gy radiation could up-regulated the PUMA expression in xenograft of human tongue squamous in naked mice. There was significant difference between the apoptosis proliferation index （PI） or index （AI） in tumors between therapy group and control groups （P＜0.01）.

Conclusions

The radiation-inducible promoters can be served as a molecular switch to upgrade PUMA expression in transplanted human tongue squamous carcinoma in naked mice， and low dose induction-radiation can significantly improve the therapeutic efficiency. It is possible to validate this strategy in the targeted treatment of tongue suqmous cell carcinoma.

Key words: Radiation-inducible promoter, p53 up-regulated modulator of apoptosis, Tongue, Carcinoma， squamous cell, Gene therapy

Xiao-hua CHEN, Fang-fang JIANG, Bin LIU, Xiao-lin WU, Dong-sheng YU. Radiation-inducible promoters mediated PUMA gene in the treatment of xenograft of human tongue squamous in naked mice[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2012, 6(05): 421-425.

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Figures/Tables 6

References 11

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基因	大小	引物序列
PUMA	1900	上游：5'-CGGAAGGTCCTCAGCCCTCGTTCT -3'
		下游：5'-AGGAGTCTCATGATGAGATGGTACA-3'
β-actin	780	上游：5'-GGTCGGAGTCAACGGATTTGGTCG-3'
		下游：5'-CCTCCGACGCCTGCTTCACCAC-3'

基因	大小	引物序列
PUMA	1900	上游：5'-CGGAAGGTCCTCAGCCCTCGTTCT -3'
		下游：5'-AGGAGTCTCATGATGAGATGGTACA-3'
β-actin	780	上游：5'-GGTCGGAGTCAACGGATTTGGTCG-3'
		下游：5'-CCTCCGACGCCTGCTTCACCAC-3'

组别	灰度值
单纯转染组	0.325±0.075
转染加放疗组	0.371±0.096

组别	灰度值
单纯转染组	0.325±0.075
转染加放疗组	0.371±0.096

组别	PI	AI
对照组	37.26±5.28	9.73±1.62
单纯放疗组	32.12±4.31	13.02±2.81
单纯转染组	32.27±5.11	14.43±4.32
放疗加转染组	22.77±3.48	27.98±5.07

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