| LIU Jianxiang,SU Xu.Construction of p-Egr-p16 plasmid and its expression induced by radiation in SMMC-7721 cells[J].Chinese Journal of Radiological Medicine and Protection,2003,23(6):439-440 |
| Construction of p-Egr-p16 plasmid and its expression induced by radiation in SMMC-7721 cells |
| Received:May 25, 2003 |
| DOI： |
| KeyWords:Egr-1 gene p16 gene p16 mRNA |
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| Abstract:: |
| Objective To construct a recombinant plasmid p-Egr- p16, investigate the expression of p16 induced by radiation in SMMC-7721 cells and its role in hepatoma suppression. Methods p-Egr-p16 vector that included Egr-1 and p 16 gene was constructed by restriction enzyme digest, ligation and introduced into SMMC-7721 cells by lipofectamine-mediated transfection. Semi-quantitati ve RT -PCR was used to detect the dose-effect and time-course manners of p16 mRNA l evel after irradiation. Results The results showed that p16 mRNA level was higher tha n that of the controls after 1-6.0 Gy irradiation. There was a peak after 2-4 Gy irradiation. p16 mRNA level increased at different times after 2 Gy irradiat ion, and the increase was marked at the 24th hour. Conclusions It is shown that p16 expression levels in SMMC-7721 c ells at 2-48 h after 2 Gy irradiation are higher than those of the sham-irradi at ion group. p16 expression levels show a time-dependant increase from 2 to 24 ho urs, and then a decrease at 48 and 72 h, but it is still higher than the sham-i rradiation group. It is suggested that 60Coγ-rays could activate Eg r-1 gene promoter and induce the expression of downstream gene p16. |
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