ABSTRACT
Background and purpose: Protein kinase P1k3 could increase the transcriptional activity of p53. However, the effect of P1k3 on the transcriptional activity of p73 is still unknown. Our study was to investigate the effect of P1k3 on the transcriptional activity of p73. Methods: Luciferase reporter assay, RT-PCR and colony formation assay were adopted to study the effect of P1k3 and kinase-deficient P1k3 (K52R) on the transcriptional activity of p73 in p53-deficient human lung carcinoma H1299 cells. Results: Luciferase reporter assay showed that p73 increased the luciferase activities induced by p21~(WAF1) and Bax promoters (P<0.05). After co-transfection with p73 and P1k3, the luciferase activities induced by p21~(WAF1) and Bax promotors were significantly decreased in a dose-dependent manner as compared with the group that transfected p73 only (P<0.05). However, kinase-deficient Plk3 (K52R) had no significant effect on the luciferase activities induced by p21~(WAF1) and Bax promoters (P>0.05). RT-PCR showed that p73 increased the mRNA expressions of p21~(WAF1) and BAX (P<0.05). P1k3 decreased the expressions of p21~(WAF1) and Bax induced by p73 (P<0.05). Kinase-deficient P1k3 (K52R) had no significant effect on the expressions of p21~(WAF1) and Bax induced by p73 (P>0.05). Colony formation assay revealed that p73 decreased the colony formation of H1299 cells (P<0.05). P1k3 decreased the inhibitory effect of p73 on the colony formation of H1299 cells (P<0.05). Kinase-deficient P1k3 (K52R) had no significant effect on the inhibitory effect of p73 on the colony formation of H1299 cells (P>0.05).Conclusion: P1k3 can inhibit the transcriptional activity of p73, where as kinase-deficient P1k3 has no effect on the transcriptional activity of p73.
ABSTRACT
Objective To investigate the genetic association between mutation in cytochrome P450 1A2 (CYP1A2) gene and Erxian suop response. Methods Forty-eight patients with male climacteric syndrome of both Yin and Yang deficiency received a treatment of erxian suop with three months. The severity of symptoms and responses to Erxian suop were quantizated by the method of Department of Psychology in Istanbul Bosphorus University and Clinical Research Guidance Principle on New Drug of Traditional Chinese Medicine published by China. A single nucleotide polymorphism (SNP) at position 2964 (G-A) of CYP1A2 gene were identified by polymerase chain reaction (PCR) in order to analyse the relationship between Erxian response and CYP1A2 single nucleotide polymorphism. Results Statistical analysis showed there was no significant difference among genetype and genetype distribution of CYP1A2 G2964A between the patients and control group. The genotype of the 16 patients among 32 effective patients was G/G (56.25%). The genotype of the 11 patients among 16 ineffective patients was G/A (68.75%). There was no significant difference in symptoms, physical sign between the two groups at the pre-treat and post-treat. Also the two groups had no significant difference among the score of major symptoms and hyposymptoms. Conclusion The patients with genotype of G/G have higher efficiency, while with genotype of G/A have lower efficiency among patients with CYP1A2 G2964A gene when treated with erxian soup.