G6PD Regulates Cyclin E1 and CDK2 to Promote Cell Proliferation and Its Prognostic Value in Clear Cell Renal Cell Carcinoma / 中国生物化学与分子生物学报

Clear cell renal cell carcinoma (ccRCC) has been proved to be a metabolic disease with high

Homoharringtonine activates ATM/p53 pathway to inhibit proliferation of human liver cancer cell PLCS / 中国药理学通报

Aim To explore the effect of homoharringtonine (HHT) on the prohferation of liver cancer cell PLCS and its possible mechanism. Methods CCK-8 and EdU were used to detect the effect of HHT on the proliferation of PLCS cells; flow cytometry was employed to assess the effect of HHT on cell cycle of PLCS; Western blot was applied to measure the expression levels of cycle-related proteins cyclinA, CDK 2, p 2 1, p53 and A T M. Results Treated with HHT (0, 5, 10, 20, 40, 80 • L

Mechanism of Fluspirilene suppressing HCC through decreasing Akt expression / 中国药理学通报

Aim To explore the mechanism of Fluspirilene inhibiting HCC through decreasing the expression of Akt.Methods The difference of mRNA was verified by the test of protein expression between Fluspirilenc treatment group and control group by HCC experiment in vivo and vitro, including Western blot, IHC after mRNA array.Results Akt expression was lower in Fluspirilene treatment group than that in control group by mRNA array.Protein expression of Akt, phosphorylate-CDK2 and phos- phorylate-Rb decreased massively in Fluspirilene treatment group in a concentration-dependent manner in HepG2 and Huh7 cells by Western blotting compared with those in control group.Declined expression of phosphorylate-Akt was proved in a concen- tration-dependent manner in xenograft tumor tissues in Fluspirilene treatment group compared with that in control group in IHC test.Conclusions Fluspirilene inhibits HCC by decreasing significantly the protein expression of Akt, phosphorylat-Akt, phos- phorylate-CDK2 and phosphorylate-Rb.

Synthesis and biological evaluation of new 2-(4-fluorophenyl) imidazol-5-ones as anticancer agents

Two series of imidazolones were designed, synthesized, and evaluated for their anticancer activity against four cancercell lines: Hela, MCF-7, PC3, and HCT-116, where four compounds 6, 25, 26, and 29 showed good potency againstthe whole panel. Compound 30 showed a cytotoxic effect against PC3 cell lines compared to that of the standarddoxorubicin with IC50 = 8.15µM, while compounds 4 and 18 showed moderate activity with IC50 range of 10.58–11.45µM. Enzyme inhibition assay was implemented against CDK2A and VEGFR-2; where varied activities were obtained.Compound 6 exhibited the highest inhibitory activity against VEGFR-2 with an IC50 value of 67 nM and moderateinhibition against CDK2A, while compound 26 achieved the best result against CDK2A with an IC50 value of 0.66 µM

Novel small-molecule CDK2-cyclinA2 inhibitors: design, synthesis, and biological evaluation / 上海交通大学学报（医学版）

Objective · To design and synthesize a series of benzenesulfonamide derivatives, test their inhibitory activity to CDK2-cyclinA2 kinase, and investigate the structure-activity relationship. Methods · Virtual screening was executed via computer-aided drug design according to the ATP binding site in CDK2-cyclinA2 protein crystal. A series of benzenesulfonamide derivatives were designed and synthesized on the basis of the interaction modes between the lead compound and the CDK2-cyclinA2. The biological evaluation of compounds was made through the CDK2-cyclinA2 in-vitro kinase activity detection system. Results · Twenty-nine new benzenesulfonamide compounds were prepared, and their inhibitory activity to CDK2-cyclinA2 was elicited. WZ-026 had the highest inhibitory parameter, which half maximal inhibitory concentration (IC50) was 3.81 μmol/L. Conclusion · By multipurpose utilization of virtual screening, chemical synthesis, and biological activity test, a benzenesulfonamide compound WZ-026 was found, which has great inhibitory activity towards CDK2-cyclinA2. Preliminary structure-activity relationship of compounds was obtained.

miR-205 promotes proliferation and invasion of laryngeal squamous cell carcinoma by suppressing CDK2AP1 expression

BACKGROUND: The aberrant expression of microRNAs (miRNAs) has been found in various types of cancer. miR-205 was reported to be upregulated in laryngeal squamous cell carcinoma (LSCC) tissues, however, the mechanisms by which miR-205 functions as a regulator of LSCC are largely unknown. RESULTS: In this study, Real-time qPCR and Western blot assay showed that expression of miR-205 was upregulated and expression of cyclin-dependent kinase 2-associated protein 1 (CDK2AP1) was downregulated in LSCC tissues. The expression levels of miR-205 were negatively related to those of CDK2AP1 in LSCC tissues and cell lines. Moreover, we found that miR-205 was the upstream regulator of CDK2AP1 and could suppress the CDK2AP1 expression in LSCC cells. 3-(4,5-dimethylthiazal-2-yl)-2,5-diphenyl-tetrazolium bromide assays and transwell invasion assay were performed to test the proliferation and invasion of LSCC cells. Gelatin zymography was used to detect the activity of MMP2 and MMP9. CDK2AP1, c-Myc and CyclinD1 expression in cells was assessed with Western blotting. We found that miR-205 was the upstream regulator of CDK2AP1 and could suppress the expression of CDK2AP1 in LSCC cells. In addition, miR-205 significantly induced cell proliferation and invasion by suppressing CDK2AP1 expression. Consistent with miR-205 inhibitors, overexpressed CDK2AP1 suppressed the activity of MMP2 and MMP9 and c-Myc and CyclinD1 expression in LSCC cells. CONCLUSION: These findings help us to better elucidate the molecular mechanisms of LSCC progression and provide a new theoretical basis to further investigate miR-205 as a potential biomarker and a promising approach for LSCC treatment.

Expressions and Clinical Significance of Runx3,Smad4,Cdk2 and p21 in Gastric Cancer / 胃肠病学

Background:Tissue microarray has been increasingly used in research of malignancies. It has been revealed that TGF-β signaling pathway contributes to the tumorigenesis and progress of malignancies. Aims:To determine the expressions of Runx3,Smad4,Cdk2 and p21,the key molecules in TGF-β signaling pathway by tissue microarray,and investigate their correlations with clinicopathological features and prognosis of gastric cancer. Methods:A total of 378 paraffin embedded tissue blocks,including 130 gastric cancer tissue and 248 para-cancer tissue from 130 patients undergoing radical resection of gastric cancer were obtained. Tissue microarray and immunohistochemistry were used to determine the expressions of Runx3,Smad4,Cdk2 and p21. Results:The aberrant expression rates of Runx3,Smad4, Cdk21 and p21 in gastric cancer tissue were significantly higher than those in para-cancer tissue(67. 7% ,35. 4% , 63. 8% and 70. 0% vs. 14. 1% ,12. 5% ,18. 1% and 37. 1% ,P < 0. 05,respectively). Aberrant expression of Runx3 was closely correlated with histological grade and lymph node metastasis of gastric cancer( P < 0. 05),while aberrant expressions of Smad4 and p21 were correlated with histological grade only(P < 0. 05);aberrant expression of Cdk2 was correlated with histological grade,lymph node metastasis and TNM staging(P < 0. 05). Pairwise correlations were seen among aberrant expressions of Runx3,Smad4 and p21 in gastric cancer,while Cdk2 was correlated with Runx3 only. Kaplan-Meier survival curve showed that 5-year survival rates in Runx3,Smad4 and p21 aberrant expression groups were significantly lower than those in normal expression groups(P <0. 05). Furthermore,Cox proportional hazard model indicated that Runx3 and Smad4 were independent prognostic factors for gastric cancer. Conclusions:Runx3,Smad4,Cdk2 and p21 might play pivotal roles in tumorigenesis and progression of gastric cancer. As interactions occurred among these four proteins,whether Runx3 and Smad4 could be used for predicting the prognosis of gastric cancer needs to be further studied.

Effects of Buyang Huanwu decoction on the expression of Cyclin D1 and Cdk2 in rats with post-stroke depression / 中华行为医学与脑科学杂志

Objective To observe the effects of Buyang Huanwu decoction(BYHW) on the expression of Cyclin D1 and Cdk2 in rats with post-stroke depression(PSD).Methods The rats model of focal cerebral ischemia was established by means of middle cerebral artery occlusion(MCAO).Then three weeks of salute-living and chronic unpredictable mild stress(CUMS) was given to the animal after cerebral stroke to induce the post-stoke depression animal model.The rats were divided into 5 groups:the sham operated group,the midge cerebral artery occlusion(MCAO) group,the PSD group,the fluoxetine group and the BYHWD group.The rats were subjected to left MCAO rebuilding in consistent focal cerebral ischemia,followed by an 21-day exposure to chronic mild stress (CMS)and single housing to induce PSD animal model.All rats were killed in 7,14 and 21 day after operation.The expressions of Cyclin D1 and Cdk2 were measured by immunohistochemical staining.Results Pathological changes such as hippocampal nerve cell regression,degeneration and necrosis were observed in the model rats compare with the sham operated group.The expression of Cyclin D1 in normal hippocampus in 7,14 or 21 day after operation was (1.16±0.34)％,(1.62±0.29)％ and (1.60±0.24)％ respectively,and Cdk2 was (1.52±0.26)％,(1.85±0.23)％ and (1.97±0.10)％.After PSD the expression of Cyclin D1 was (49.69±5.68)％,(58.17± 2.89) ％ and (50.87 ± 2.48) ％ respectively,and Cdk2 was (50.63 ± 2.93) ％,(70.34± 1.47) ％ and (61.35 ± 3.04) ％.Compared with model group,Fluoxetine and BYHW significantly reduced the numbers of Cyclin D1 and Cdk2 positive cells,the expression of Cyclin D1 was (16.62±4.27)％,(29.66±5.24)％ and (26.71±1.32)％ at fluoxetine group,and Cdk2 was (18.05±2.26) ％,(33.84±3.12) ％ and (24.51±2.66) ％.The expression of Cvclin D1 was (14.62±2.06)％,(26.89±3.41)％ and (23.68±2.01)％ at BYHWD group,and Cdk2 was (16.60± 2.42) ％,(20.09±3.45) ％ and (22.19± 1.70) ％.Conclusion The abnormal expression of Cyclin D1 and Cdk2 at the PSD rats indicate that they may be involved in the mechanism of neuronal death.Buyang Huanwu decoction may reduce the neuronal apoptosis through down-regulating the expression of Cyclin D1 and Cdk2.

Effects of DCA on Cell Cycle Proteins in Colonocytes

PURPOSE: Evidence that indicates bile acid is a promoter of colon cancer exists. Deoxycholic acid (DCA) modifies apoptosis or proliferation by affecting intracellular signaling and gene expression. However, because previous studies have been based on studies on colon cancer cell lines, the effect of DCA on normal colonocytes is unknown. METHODS: Normal colonocytes and Caco-2 and HCT116 cells were treated with 20 micrometer and 250 micrometer of DCA, and the effect of different concentrations of DCA was measured based on the expression of cell-cycle-related proteins by using Western blots. RESULTS: The expressions of CDK2 and cyclin D1 for different concentrations of DCA in normal colonocytes and colon cancer cells were similar, but the expressions of cyclin E and A were significantly different. In HCT116 colon cancer cells, the expression of cyclin E increased regardless of the DCA concentration, but in normal colonocytes and Caco-2 cells, the expression of cyclin E was not changed or decreased. In HCT116 colon cancer cells, the expression of cyclin A was not changed or decreased regardless of the DCA concentration, but in normal colonocytes and Caco-2 cells, the expression of cyclin A was increased at a DCA concentration of 20 micrometer. CONCLUSION: The effect of DCA on stimulating cell proliferation suggests that DNA synthesis is stimulated by an increased expression of cyclin E in colon cancer cells. Our results suggest that a low dose of DCA induces cellular proliferation through increased expression of cyclin A and that a high dose of DCA induces decreased expression of cyclin E and CDK2 in normal colonocytes.

Expressions of cyclinE,cdk2 and p27~(kip1) in invasive pituitary adenomas and the clinical significance / 重庆医科大学学报

Objective:To study the expressions of cyclinE,cdk2 and p27kip1 in invasive pituitary adenomas and their relation to clinical biological behaviors. Methods:Western blotting analysis was adopted to detect the expressions of cyclinE,cdk2 and p27kip1 in 37 cases of noninvasive pituitary adenomas and 21 cases of invasive pituitary adenomas. Results:The expressions of cyclinE and cdk2 in invasive pituitary adenomas were significantly higher than those in noninvasive pituitary adenomas(both P

Relationship between the expression levels of cell cycle regulators and the proliferation ability of human giant-cell lung carcinoma cell strains 95C and 95D / 中国癌症杂志

Purpose:To study the molecular mechanism that d etermines the cell cycle differences between human giant-cell carcinoma cell st rains 95C and 95D. Methods:FACscan was used to analyze the cell cycle of 95C and 9 5D cells, and the proliferation ability of 95C and 95D cells was measured by MTT assay. We further detected the expression levels of cell cycle factors by Weste rn blotting. Results:Our data showed that the proliferation ability of 95D c ells is higher than that of 95C cells, and that the cell number in S phase of 95 D cells is much larger than that of 95C cells. We further found that the express ion level of p27 in 95D cell is lower than that of 95C cells. In addition, the e xpression levels of CDK2 and p-RB of 95D cells are higher than that of 95C cell s. Conclusions:Our results indicated that low expression level of p27 lead to the increased proliferation ability of 95D cells, which might reflec t the progression of human lung cancer cells in cell cycle facets.