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Objective: To investigate the effects of SPAG6 silencing and decitabine on apoptosis and phosphatase and tensin homolog (PTEN) methylation in SKM-1 cells in vitro and in vivo. Methods: SKM-1 cells were transfected with a lentiviral vector to silence the expression of SPAG6. Cell survival rate was detected by CCK8 after treatment with decitabine, and cell apoptosis was detected by flow cytometry. Protein expression and methylation of PTEN were detected using Western blot and merozoite surface protein (MSP) . An non-obese diabetic/severe combined inmunodeficiency disease (NOD/SCID) mice xenograft tumor model was established, and the apoptosis and PTEN expression of tumor tissue were observed through terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and immunohistochemistry (IHC) , respectively. Results: After lentivirus transfection, SPAG6 in the interference group was silenced successfully. CCK8 results indicated that the cell survival rate of SKM-1 cells treated with decitabine decreased. Flow cytometry showed that the apoptosis rate of cells treated with decitabine [ (17.35±3.37) %] was higher than that of the untreated group (5.09%±2.06%) and the apoptosis rate of the SPAG6 silencing combined with the decitabine treatment group was the highest [ (36.34±4.00) %]. After treatment with decitabine, the expression of DNMT1 decreased, while the expression of PTEN increased, and the promoter methylation degree of PTEN also decreased. Moreover, the increased protein level caused by PTEN demethylation was the most obvious in the SPAG6 in the interference shRNA group treated with decitabine. In NOD/SCID mice, the tumor volume of the decitabine group was significantly smaller than that of the placebo group, and the tumor volume of the SPAG6 silencing combined with the decitabine treatment group was the smallest. Additionally, the apoptosis rate was the highest (the positive ratio was 3.57±0.48) . Conclusion: SPAG6 silencing may enhance the apoptosis level and the effect of PTEN demethylation in SKM-1 cells and enhance the antitumor effect of decitabine in the NOD/SCID xenograft mouse model.

Objective: To identify the therapeutic efficacy of lithium chloride (LiCl) on phosphatase and tensin homolog deleted on chromosome ten (PTEN)-deficient tumors. Methods: First, the Catalogue of Somatic Mutations in Cancer for mutation spectrum of human endometrial carcinoma samples was analyzed. Second, the relationship between PTEN abundance and LiCl inhibition of endometrial cancer cell lines using Pten(+/+) and Pten(-/-) mouse embryonic fibroblast (MEF) lines was investigated. Moreover, potential alterations of mammalian target of rapamycin (mTOR) signaling pathway after treatment with LiCl were checked.Last,LiCl's efficacy on PTEN null tumors was studied. Results: PTEN was mutated in 39% of endometrial carcinomas. LiCl preferentially inhibited the proliferation of PTEN-deficient endometrial carcinoma cells and MEFs. Furthermore, LiCl blocked PTEN-deficient tumor development. Mechanistically, LiCl down-regulated mTOR signaling. Conclusions: PTEN is the most frequently mutated gene in endometrial carcinoma.By targeting mTOR signaling pathway,LiCl is a promising regimen for the treatment of tumors with PTEN deficiency.

Objective: To observe the changes of phosphatase and tensin homolog deleted on chromosome ten(PTEN)/ phosphatidyl Inositol 3-kinase(PI3K)/ vascular endothelial growth factor(VEGF)signaling pathway in a rabbit Kawasaki disease model., Methods: Model of Kawasaki disease was established in weanling Japanese big-eared rabbits with 10% bovine serum venous injection (2.5 ml/kg, 2 times, and 2 week's interval) through the ear. Twenty four rabbits were divided into 4 groups: control group (without injection of 10% bovine serum albumin, six rabbits); 1 day group (sacrificed a the second day after the establishment of Kawasaki disease models, six rabbits); 7 day group (sacrificed at the seventh day after establishment of Kawasaki disease model, six rabbits); 30 day group (sacrificed at the thirtieth day after establishment of Kawasaki disease model, six rabbits). Pathological analysis was performed on coronary artery tissue samples. The express of PTEN and PI3K were detected by immunohistochemistry. The levels of VEGF and CK were also examined with ELISA and white blood cells were counted., Results: (1) Coronary artery of model groups was thinner, distorted and had enlarged lumen. (2) PTEN expression in 1 d group, 7 d group and 30 d group were 58.5 ± 12.9, 73.2±9.9 and 109.6 ± 24.4, respectively, significantly higher than in the control group (25.5 ± 6.9, P<0.01 or 0.05). (3) The express of PI3K was significantly upregulated in 1 d group(57.2±11.1)and 7 d group(39.9±4.8) compared to control group(19.1±3.5, P<0.01 or 0.05). The expression level of PI3K in 30 d group was 18.8 ± 7.5, which was similar as control group (P>0.05) and significantly lower than 1 d and 7 d group (both P<0.05). (4) Similarly, the level of VEGF in 1 d group, 7 d group ((89.1 ± 15.5) ng/L, (76.9±9.9) ng/L) were significantly higher while it was significantly lower in 30 d group ((19.8 ± 4.4)ng/L) compared with the control group ((33.9 ± 6.7) ng/L, P<0.01 or 0.05). The level of VEGF in 7 d group was significantly lower than in 1 d group (P<0.05) and the level of VEGF was significantly lower in 30 d group than in 7 d group (P<0.01). (5)Creatine kinase levels were significantly higher in 30 d group than in control group (P<0.05) and there were no significantly different between control group, 1 d group and 7 d group(all P>0.05). (6)White blood cell count were significantly higher in 1 d group, 7 d group and 30 d group than in control group (all P<0.01)., Conclusion: The level of PTEN/PI3K/VEGF signaling pathway change after establishment of rabbit Kawasaki disease model and the signaling pathway might be involved in this model.

Objective: To explore the correlation between the expression of C-terminal tensin-like protein (CTEN) and the prognosis of hepatocellular carcinoma (HCC)., Methods: Using immunohistochemistry, we detected CTEN protein level in samples of primary lesion and adjacent non-tumor lesion collected from 240 patients with HCC. The relationship between CTEN expression and clinicopathology, 5 year recurrent-free survival, or overall survival was evaluated by Chi-square test, Kaplan-Meier, or Cox regression analysis., Results: High CTEN expression was detected in 55% of hepatocellular carcinoma tissues and 20% of adjacent carcinoma tissues (P< 0.001). CTEN expression was positively correlated with tumor diameter (P=0.022), venous invasion (P=0.007) or TNM stages (P=0.022). Five-year recurrence-free survival time (P< 0.001) and overall survival time (P< 0.001) in patients with high CTEN expression were significantly less than those in patients with low CTEN expression. Multivariate Cox regression analysis revealed that the CTEN expression was an independent prognostic marker for HCC (all P< 0.05)., Conclusion: CTEN protein may play a role in the genesis and development of HCC, and it can function as a prognostic marker.

Objective: To investigate the role of mutation and mRNA expression of tumor suppressor gene phosphatase and tensin homolog deleted in chromosome 10 (PTEN) in tumorigenesis and progression of oral squamous cell carcinoma (OSCC)., Methods: The mutation of exon 3, 5, 6 and exon 8 of PTEN gene in 42 oral squamous cell carcinoma tissue and paired normal tissue were detected by polymerase chain reaction (PCR) and DNA sequencing methods. The levels of PTEN mRNA expression in these tissues were assayed using semi-quantitative RT-PCR. Data was analyzed with SPSS 14.0 software package., Results: Mutated exon 5 of PTEN gene was found in 2 cases of advanced OSCC. The expression of PTEN mRNA was detected in all OSCC and paired normal tissue. The level of PTEN mRNA in OSCC tissue (0.36 +/- 0.12) was significantly lower than that of paired normal tissue (0.64 +/- 0.09, P < 0.01)., Conclusions: The decreased expression of PTEN mRNA contributes to tumorigenesis of OSCC.

Aim: To investigate the expression changes of tensin in human kidney mesangial cells with stimulating of high concentrations of glucose (HG)., Methods: Human mesangial cells were cultured and stimulated with different concentrations (5 mmol/L, 30 mmol/L) of glucose for 48 h, 72 h and 5 d . The expression changes of tensin was detected by immunofluorescence technique, and fibronectin of supernatant were detected by ELISA., Results: The expression of tensin in mesangial cells was more increased significantly with stimulating of high concentrations of glucose with different culturing time, and fibronectin was secreted into the supernatant and the concentrations were increased with culturing time., Conclusion: The expression of tensin is increased in mesangial cells with stimulating of high concentrations of glucose, and tensin plays an important role in pathogenesis of accumulation of extracellular matrix.