Effect and related mechanism of decitabine on mantle cell lymphoma cell line Mino / 白血病·淋巴瘤

Objective To investigate methylation of the P15INK4B and p27kipl genes in human mantle cell lymphoma cell line Mino,to evaluate the effects of decitabine on demethylation of p15INK4B and p27kip1 genes and on apoptosis of Mino cells and its relative mechanism. Methods Mino cells were after treated with various concentration of decitabine, the cell viability, cell cycle distribution or the apoptosis of Mino cells was respectively analyzed by trypan dye-exclusion assay or flow cytometry.The mRNA and protein expression of p15INK4B 、p27kip1 and bcl-2 were studied by RT-PCR or Western blot, respectively.Methylation of the p15INK4B and p27kip1 genes in Mino cells were determined by PCR using the methylation specific primer(MSP).Results Decitabine significantly inhibit the cell growth,induced G1 arrest and promoted apoptosis of Mino cells. The expression of p15INK4B and p27kipl mRNA were both significantly increased,wheres bcl-2 mRNA was decreased, After treatment with 6.4,3.2,1.6 mmol/L decitabine for 72 h,the methylationg of p15INK4B gene were decreased to 25.2 ％ 、48.2 ％ and 65.0 ％respectively,in the meantime,the methylationg of p27kip1 gene was decreased to 20.21％ 、50.2 ％ and 70.0 ％. Conclusion The hp15INK4B and p27kip1 genes of Mino cells are methylated and down-regulated.Decitabine can inhibit the proliferation and induce the apoptosis of Mino cell lines,with the reduction of bcl-2 gene and demethylation of p15INK4B and p27kip1 genes.

Effect of Helicobacter pylon on gastric epithelial cells proliferation, apoptosis and expression of p27kipl protein in vitro / 中华微生物学和免疫学杂志

Objective To explore the influence of Helicobacter pylori (Hp) on the proliferation, apoptosis and expression of p27kipl protein of gastric epithelial cells in vitro. Methods SC, C-7901 cell pro-liferation was examined by flow cytometry after incubation with different concentration of NCTC11637. The effect of NCTC11637 on cell apoptosis was also evaluated by flow cytometryo And the expression of p27kipl of gastric epithelial cells was detected by immunohistochemical analysis and in situ hybridization, respectively. Results Cell proliferation was enhanced when co-incubated with the Hp in low concentration (3.4 x 104 to 1.9 x 105 CFU/ml) but inhibited in higher concentration (≥4.8×106CFU/ml). However, cell apoptosis was increase when co-incubated with the Hp in high concentration (≥9.6 ×105 CFU/ml) showing concen-tration dependent picture. In addition, the co-incubation of SGC-7901 with Hp led to decrease of the expres-sion of p27kipl protein but not mRNA in a Hp concentration dependent way. Conclusion Hp could effect the gastric epithelial cells apoptosis and proliferation directly and influence the expression of p27kips protein which might facilitate gastric carcinoma.

Effect of p27Kipl Inhibition on Proliferation of Bovine Corneal Endothelial Cells by RNA Interference / 华中科技大学学报（医学）（英德文版）

Three plasmids (pGenesil-P1, pGenesil-P2, pGenesil-P3) with different p27Kipl-shRNA sequences were designed and synthesized. Their effects on the proliferation of bovine corneal endo- thelial cells (bCEC) were investigated. Plasmid expressing irrelevant shRNA with a random combi- nation was used as negative control (pGenesil-HK). The recombination of four plamids was con- firmed by restrictive enzyme digestion and sequence analysis. The expression of mRNA and protein of p27Kipl was detected by RT-PCR and Western blotting after stable transfection. The expressions of p27Kipl mRNA and p27Kipl protein of pGenesil-P1 group, pGenesil-P2 group and pGenesil-P3 group were all lower than those in the pGenesil-HK group and the blank group (non-transfected group), pGenesil-P3 had the strongest inhibitory effect and was selected for the next steps. The pro- liferation rates of the pGenesil-P3 group, the pGenesii-HK group and the blank group were assessed by MTT. The influence of shRNA-p27Kipl on bCEC cell cycle was detected by flow cytometry (FCM). Compared with the control groups, the proliferation rate of the pGenesiI-P3 group was increased significantly, and the ratio of S-phase also increased. It is concluded that shRNA-p27Kipl could down-regulate the expression of p27Kipl effectively and increase the proliferation of bCEC. RNA interference (RNAi) may be an effective means to promote the proliferation of CEC.

The Clinical Meaning Of P～(27kipl)and carcinoma of the pancreas / 医学研究杂志

P～ 27 gene that has been found in recent years can inhibit cancer.It codes P～ 27 protein named CDKI.CDKI plays an important role in regulating the cell cycle.The symptom of carcinoma of the pancreas in early stage isn't distinctiveness.Once it is found,it has been in later stage.To explore the biological meaning of P～ 27kipl and carcinoma of the pancreas,we measure the expression of P～ 27kipl protein in carcinoma of the pancreas and normal pancreas tissue by using the method of immune tissue chemistry SP.

Expression and implication of Survivin and its correlation with p27～(kipl) in gallbladder cancer / 中国现代普通外科进展

Objective:To study the expression of Survivin and p27～(kipl)and their relationship with the clinical features of gallbladder cancer.Methods:Fifty specimens of gallbladder cancer,20 specimens of chronic cholecystitis,20 specimens of polypoid lesions of gallbladder(PLG)were included,the expression of Survivin and p27～(kipl)were detected using immunohistochemistry assay.Results:In 50 cases of gallbladder cancer,39 cases expressed Survivin and 22 cases expressed p27～(kipl).There was a negative correlation between the expression of p27 and survivin(P0.05),but the expression of p27～(kipl)was related to the pathological grade,clinical stage and lymph node metastasis of gallbladder cancer(P

Expression of P27~kipl and mutant P53 protein in thyroid tumors / 第二军医大学学报

Objective:To study the expression of P27kipl and mutant P53 protein in thyroid tumors, and to understand the relationship between the biological behavior and prognosis of benign and malignant thyroid tumors. Methods: We used S-P immunohistochemical technique to study the expression of P27kipl and mutant P53 protein in 60 cases of thyroid tumors, including 40 cases of malignant tumors and 20 cases of benign tumors. Categorical data were analyzed by using X2 tests. All reported significance levels were two-tails. The software system used for statistical analysis was SPSS10. 0. Results: In benign and malignant tumors, P27kipl protein positive expression was detected in 18 of 20 benign cases (90. 0%) and 24 of 40 malignant cases (60. 0% ) respectively(P P

Reduced Expression of p27 Correlates with High Grade Malignancy and is Associated with Poor Clinical Outcome in Human Greast Cancer / 대한암학회지

PURPOSE: The cyclin-dependent kinase inhibitor p27Kipl is a negative regulator of cell proliferation. Its expression is known to be altered in proteasome-dependent manner without changes in DNA level. Reduced expression of p27Kipl is associated with aggressive behavior in a variety of human cancers. We investigated expression of p27Kipl protein in human breast cancer using immunohistochemistry to assess its biologic implication along with cell cycle analysis by flow cytometry. MATERIALS AND METHODS: We peformed immunohistochemical assay for expression of p27 in total 68 patients with invasive ductal cancer along with 27 benign ductal hyperplasia and 10 ductal carcinoma in situ. The data were analyzed in association with proliferative activity of the same tissues and biologic parameters. RESULTS: In epithelial cells of normal and benign breast disease, expression of p27Kipl was well preserved while its expression markedly decreased in breast cancer (45 of 68). Expression of p27Kipl was significantly reduced in poorly differentiated cancers and in advanced stage of the disease. Levels of p27Kipl expression correlated with cell populations in GO/Gl phase of the cell cycle. In survival analysis, p27Kipl was useful to predict disease-free survival but not overall survival of the patients after adjuvant chemotherapy. CONCLUSION: p27Kipl seems to have a role in cell proliferation and differentiation process during carcinogenesis of breast cancer. The results of present study suggest that p27Kipl can be used in predicting response to systemic chemotherapy in a subset of patients with breast cancer.

Adenovirue-mediated human p27kip1 gene transfection inhibits neointimal proliferation of rabbit carotid artery after balloon injury / 中国介入心脏病学杂志

Objective To study the effects of adenovirue-mediated p27kipl gene and its protein product overexpression on neointimal proliferation of rabbit carotid artery after balloon injury. Methods After rabbit arterial carotid injuried, injuried arterial segments were immediately infected by LacZ recombinant adenovirues (AdLacZ) and human p27kipl recombinant adenovirues (Adhp27kip1) in vivo, respectively. Western blot, x-gal stainning, HE stainning, immunochemistry and compute image system were used to analyze the expression of exogenous p27kipl gene and its protein product and the effects on neointimal proliferation. Results Comparison to AdLacZ infected or uninfected arterial segments, Adhp27kip1 infected segments overexpressed p27kipl protein, the peak of expression was in 7-14 d, expression lasted more than 4 weeks. After 4 weeks, in uninfected, AdLacZ infected or Adhp27kipl infected segments, the neointimal area was 1.106 mm2, 0.988 mm2 and 0.278 mm2, respecively; the rate of luminal stenosis was 87.07%, 65.40% and 32.14%, respectively. Conclusion Exogenous p27kipl gene and its protein product overexpression in injuried arterial segments could inhibit neointimal proliferation and luminal stenosis significantly after artery injury.

Expression of P27kipl protein and cyclin E protein in renal cell carcinoma / 重庆医科大学学报

Objective:To investigate the expressions of P27kipl (P27) protein and cyclin E protein in human renal cell carcinoma and in paracarcinoma kidney cortex tissue,and to explore the correlation of these protein expressions with biological behavior of tumors.Methods:P27 and cyclin E expressions were studied in formalin-fixed,paraffin-embedded sections of renal cell carcinoma and in paracarcinoma kidney cortex tissue by means of a specific and sensitive immunohistochemical S-P assay.Results:The positive rates of P27 protein and cyclin E protein in renal cell carcinoma were significantly higher than those in paracarcinoma kidney cortex tissues ( P