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. 2004 Jul 1;10(13):1979-83.
doi: 10.3748/wjg.v10.i13.1979.

Reversal of 5-flouroucial resistance by adenovirus-mediated transfer of wild-type p53 gene in multidrug-resistant human colon carcinoma LoVo/5-FU cells

Zhi-Wei Yu  1 , Peng Zhao, Ming Liu, Xin-Shu Dong, Ji Tao, Xue-Qin Yao, Xin-Hua Yin, Yu Li, Song-Bin Fu
Affiliations

Reversal of 5-flouroucial resistance by adenovirus-mediated transfer of wild-type p53 gene in multidrug-resistant human colon carcinoma LoVo/5-FU cells

Zhi-Wei Yu et al. World J Gastroenterol. .

Abstract

Aim: To observe the reversal effects of wide-type p53 gene on multi-drug resistance to 5-FU (LOVO/5-FU).

Methods: After treatment with Ad-p53, LOVO/5-FU sensitivity to 5-Fu was investigated using tetrazolium dye assay. Multidrug resistance gene-1 (MDR1) gene expression was assayed by semi-quantitative reverse transcription-polymerase chain reaction and the expression of p53 protein was examined by Western blotting.

Results: The reversal activity after treatment with wide-type p53 gene was increased up to 4.982 fold at 48 h. The expression of MDR1 gene decreased significantly after treatment with wide-type p53 gene, and the expression of p53 protein lasted for about 5 d, with a peak at 48 h, and began to decrease at 72 h.

Conclusion: Wide-type p53 gene has a remarkable reversal activity for the high expression of MDR1 gene in colorectal cancers. The reversal effects seem to be in a time dependent manner. It might have good prospects in clinical application.

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Figures

Figure 1
Figure 1
Effect of wild-type p53 on the growth rate of LoVo/ 5-FU cells.
Figure 2
Figure 2
Western blot analysis of wild-type p53 in LoVo/5-FU cell line and LoVo/5-FU cells transiently transfected with p53 expression vector (Ad-p53). Lane 1: LoVo/5-FU cell line. Lanes 2, 3, 4, 5, 6: LoVo/5-FU cells transiently transfected with Ad-p53 for 2, 3, 4, 5, 6 d accordingly. Levels of wild-type p53 protein (lanes 2-6) were determined by Western-blot analysis with an antibody which could detect wild-type forms of p53 protein.
Figure 3
Figure 3
Results of RT-PCR for expression of MDR1 mRNAs in p53-null and wild-type p53 expressing LoVo/5-FU cells. M: marker; Lanes 1 and 2: results of untransfected LoVo/5-FU cells; Lanes 3 and 4: results of transfected LoVo/5-FU cells.
Figure 4
Figure 4
Results of RT-PCR for expression of MDR1 mRNAs of transfected LoVo/5-FU cells at different time. M: marker; Lanes 1, 2, 3, 4, 5: the cells transfected with Ad-p53 for 2, 3, 4, 5, 6 d accordingly.
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