| Objective: The purpose of this study is to probe into the function of 14-3-3βin tumorigenesis and development of KS, and the possible mechanism in KS pathogenesis by using molecular biology and cell biology methods.Methods: We Collected tumour tissues and vicinal normal skin tissues from 21 cases of KS patients in xinjiang, and collected normal skin tissues from 12 cases of non-KS patients as control, then applied Real-time PCR and immunohistochemical technique to detect 14-3-3βmRNA and protein expression in these samples; we constructed eukaryotic expression vector of 14-3-3βby using genetic recombination technique, then transfected recombinant into Human umbilical vein endothelial cell line (ECV-304) and BC-3 cells by using liposome, the expression of 14-3-3βwas detected by western-blot after 48 hours, then observed the morphologic change, Cell Counting Kit-8(CCK-8) was used to detect the growth rate of transfected cells; Cell proliferation index(PI) was detected by immunohistochemical Envision technique, cell apoptotic index(AI) was detected by the in situ end labeling(TUNEL) method.Results: (1) The 14-3-3βmRNA expression in tumor tissues of KS patients is 3.60±2.62×10-1, the minimum and maximum of 14-3-3βmRNA expression in tumor tissues of KS patients are 0.78×10-1 and 6.83×10-1, respectively; the 14-3-3βmRNA expression in normal skin tissues of KS patients is 0.95±0.50×10-1, the minimum and maximum of 14-3-3βmRNA expression in normal skin tissues of KS patients is 0.22×10-1 and 1.82×10-1, respectively.14-3-3βmRNA expression in tumor tissues of KS patients are higher than that in normal skin tissues, the difference has statistical significance(t=2.554, P<0.05). (2) 14-3-3βprotein positive expression rate in KS was significantly higher than that in normal skin tissues(x2 =18.958, P<0.01), the difference of positive reaction degree between KS and normal skin tissues has statistical significance(t=15.136,P<0.01). (3)The eukaryotic expression vector of 14-3-3βwas successfully cloned, after the recombinant was transfected into ECV-304 and BC-3 cells for 48 hours, we detected the expression of 14-3-3βprotein, the result shows the growth rate of ECV-304 after transfected 10μg/μl, 20μg/μl, 40μg/μl, 80μg/μl recombinant were 1.24%,17.31%,27.45%,46.82%;the growth rate of BC-3 were 39.06%,45.03%,51.21%,80.35%,they have significant difference comparing to negative control group and the group that didn't transfect recombinant(p<0.05). (4) PI and AI in KS were significantly higher than that in normal skin tissues(t=6.914,3.183,P<0.01);14-3-3βwas positively correlated with PI(r=0. 671, P<0. 01) and AI(r=0.649,P<0. 05).Conclusion: (1) 14-3-3βmRNA and protein expression in tumor tissues of KS patients are higher than that in normal skin tissues, (2)the overexpression of 14-3-3βpromote the malignant proliferation of KS; (3) 14-3-3βgene promotes the proliferation function more remarkable in HHV-8 infected cells; (4) 14-3-3βgene is related with the tumorigenesis and development of KS, contribute to the proliferation and apoptosis of KS.
|
PDF Full Text Request