Effects Of Qilan Granulates On Expression Of Survivin,Bcl-2 And P53 In 4NQO-induced Rat Tongue Carcinogenesis And Their Relationship

Objective: In this study, the effects of Chinese herbal medicine Qilan Granulates on expression of Survivin,Bcl-2 and p53 was examined in 4-nitro-quinoline 1-oxide(4NQO)-induced rat tongue carcinogenesis. The study was designed to investigate these apoptotic genes in relation to cancer progression in rat and to assess the effect of Qilan Granulates.Methods: A total of 150 Sprague-Dawley (SD) rats were divided into five groups: one carcinogenesis model group (group A), three administration subgroups (group B, C, D) and one normal control group (group E). Rats in group A, B, C, D were fed with 0.002% 4NQO dissolved in drinking water to induce tongue carcinogenesis in rats. Different concentration of the herb Qilan Granulates was given to the rats during oral carcinogenesis. Rats were sacrificed at 9, 18,27and 36 weeks respectively from the beginning of the experiment. The tongues of the rats were dissected for gross and histological assessment. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was employed to detect the expression of Survivin mRNA and two-step immunohistochemistry was also employed to examine the expression of Survivin protein,Bcl-2 protein and p53 protein in the specimens.Results:1. Rats in group B, C, D had white shiny fur, lively and active. While rats in group A had yellow rough fur, sleepy and tired. They became thinner and thinner, and their weights were lighter than that of other groups of rats in the same period(P <0.01).2. Compared to the model group, the occurrences of dysplasia in three administration subgroups at 27,36 weeks were suppressed by Qilan Granulates(P <0.05),but the occurrences of dysplasia at 9,18 weeks has no significant difference (P > 0.05).3.(1)In group A the expression of Survivin mRNA:tongue cancer tissue(0.684±0.311)> dysplasia tissue (0.341±0.306)> normal tissue (0.034±0.073)(P <0.05).(2) In group A the expression of Survivin protein:tongue cancer tissue>dysplasia tissue> normal tissue(P <0.05).(3) In group A the expression of Bcl-2 protein and p53 protein:tongue cancer tissue>dysplasia tissue> normal tissue(P <0.05).4. The expression of Survivin protein in group A was positively correlated with that of Bcl-2 protein (r = 0.699, P <0.001) and that of p53 protein (P = 0.001, r = 0.419).5.(1)The expression of Survivin mRNA in tongue dysplasia (0.341±0.306) and cancer (0.684±0.311) tissue from rats in group A was significantly higher than that of from group B (0.132±0.174), group C (0.116±0.167) and group D (0.133±0.181) (P <0.05).(2) The expression of Survivin protein in tongue dysplasia and cancer tissue from group A was significantly higher than that of from group B, group C and group D (P <0.05). (3) Compared to group A, the expression of Bcl-2 protein and p53 protein in tongue dysplasia tissue of three administration subgroups was significantly lower (P <0.05), but in tongue cancer tissue, there was no significant difference between group A and three administration subgroups (P> 0.05).Conclusion:1. The weights of rats in group B, C, D were heavier than that in group A. It showed that Qilan Granulates could keep fit.2. High expression of Survivin mRNA and Survivin protein in tongue dysplasia and cancer tissue indicated that Survivin may be associated with oral carcinogenesis. High expression of Bcl-2 protein and p53 protein in tongue dysplasia and cancer tissue implied that both Bcl-2 and p53 may play an important role in oral carcinogenesis. 3. Both two-step immunohistochemical assay and RT-PCR can be used to detect the targeted genes. Compared to RT-PCR, the immunohistochemical assay was easier to perform and more cost-effective, therefore immunohistochemistry can be used in the initial detection of the targeted genes.4. The over expression of Survivin, Bcl-2 and p53 proteins found in this study indicated that a synergistic effect among these three proteins in oral carcinogenesis might be presumed.5. Qilan Granulates significantly reduced the incidence of tongue dysplasia and carcinogenesis; this suggested the potential of Qilan Granulates against malignant tumors on oral experimental carcinogenesis.6. Qilan Granulates may induce down-regulation of Survivin, Bcl-2 and p53 proteins to block the progression of carcinogenesis in dysplasia tissue, and regulated the expression of Survivin to inhibit the carcinogenesis.