Molecular mcchanism of deguelin-induced apoptosis and cell cycle arrest in lung cancer cells in vitro / 中国药理学通报

Aim To explore whether deguelin can block cell cycle and cell migration, inhibit the proliferation of non-small cell lung cancer cells.Methods H1299 cells were treated with 1.562 5, 3.125, 6.25, 12.5, 25, 50 μmol·L-1 deguelin for different time(24, 48, 72 h); cell viability was detected by CCK-8 assay, and cell migration ability was tested by scratch assay.H1299 cells were treated with 1.5, 3, 6 μmol·L-1 deguelin for 24 h.Flow cytometry with PI single staining and Annexin V-FITC/PI double staining experiment were used to evaluate cell cycle and apoptosis.qPCR was used to detect the regulatory effects of deguelin and its carbamate derivative on the Cyclin D-CDK4/6 complex at the gene level.Results Deguelin inhibited cell growth and the IC50 value of deguelin was(5.47±0.97),(4.01±0.45),(2.86±0.19)μmol·L-1 when treated with 24, 48, 72 h respectively.Deguelin also inhibited the healing ability of H1299 cells and the migration of H1299 cells significantly(P<0.05).Deguelin could block H1299 cell cycle in G1 phase.Flow cytometry combined with Annexin V-FITC/PI double staining showed that deguelin could induce apoptosis of H1299 cells.qPCR experiments showed that deguelin could down-regulate the expression of CDK4, CDK6 and Cyclin D1 genes significantly(P<0.05).Conclusions Deguelin may regulate cell cycle by down-regulating CDK4, CDK6 and Cyclin D1 genes in the cell cycle regulation system, and reduce the migration ability of tumor cells to induce apoptosis.

Effect of metformin on the expression of Cyclin Dl in human gastric cancer Cells / 国际外科学杂志

Objective To investigate the effect of metformin on human gastric cancer line SGC-7901 in vitro, trying to explore the mechanism involved. Methods Human gastric cancer SGC-7901 cell was cultured in vitro, MTT was used to study the effects of metformin on cell growth with different concentration or different time. Western blot was used to investigate the influence of metformin on the expression of Cyclin Dl in cell line . Results The result of MTT showed human gastric cancer SGC-7901 cell was cultured in vitro and was exposed to metformin in different concentration (50 mmol/L, 100 mmol/L) for 24 h,48 h,72 h. The inhibitory rates of 50 mmol/L metformin on the effects of metformin on cell growth was 32.93% ,48.64% and 61.40% and the inhibitory rates of 100 mmol/L metformin on the effects of metformin on cell growth was 35.34% , 75.44% and 88.30%. The proliferation of SGC-7901 cell was inhibited by different concentration (50 mmol/L, 100 mmol/L) metformin and,the inhibitory rates were significantly decreasing compared with the control group(P < 0.05).The inhibitory rate treated with 100 mmol/L metformin was significantly decreasing compared with 50 mmol/L metformin group (P < 0.05). The inhibitory rate treated with metformin at different time(24 h,48 h, 72 h) was significantly different(P <0.05). Metformin could inhibit the proliferation of SGC-7901 cells, with dose and time-related effects. Cyclin Dl expressed in high level in SGC-7901 and metformin significantly decreased the expression of Cyclin Dl, with dose and time-related effects. Conclusion metformin could inhibit the proliferation and lower the expression of Cyclin Dl in human gastric cancer line SGC-7901 in vitro.

Study of the expression of the cyclin D1 in chronic myelogenous leukemia / 白血病·淋巴瘤

Objective To examine the expression levels of cyclin Dl in the patients with chronic myelogenous leukemia (CML), and evaluate the pathogenesis and clinical significance of cyclin Dl in CML Methods The real-time quantitative polymerase chain reaction (RQ-PCR) was performed to detect the expression levels of cyclin Dl in the bone marrow samples of 18 patients with CML, and 16 samples of benign hemopoietic patients. The relationship between the expression levels of cyclin Dl and the progression and prognosis of patients with CML were analyzed. Results The level of cyclin Dl was higher expressed in 18 patients with CML than the control group (P <0.001). The levels of cyclin Dl was apparently higher expressed in accelerated phase /blast crisis phase than in chronic phase (P <0.05). And the RQ-PCR method showed the tendency that a significant increase was observed in the levels of cyclin Dl from 0.1980 in control group to 1.4002 in chronic phase and 5.4540 in accelerated phase /blast crisis phase. Conclusion The cyclin Dl overexpressed in CML, the roles of cyclin Dl in CML might be an oncogene expressed. The expression level is correlated with the progression and prognosis of patients with CML.

Expression of survivin and cyclin D1 protein in human non-small cell lung carcinoma / 医学研究生学报

Objective:To research the expression of survivin and cyclin D1 in human non-small cell lung cancer, and to illustrate their relationship in NSCLC. Methods:Forty-five NSCLC paraffin embe-ded samples were collected. Survivin and cyclin D1 were tested by immunohistochemical S-P method. Results:No survivin expression was present in normal lung tissues. The positive rate of survivin in NSCLC was 60% (27/45). By statistic analysis, the significant differences were found in different pathological grading and clinical phased lymph node involvement. The patient' s gender, age and histological classification were not related with the expression of survivin. Conclusion:Survivin may play an im-portment role in the process of carcinogenesis and development of NCLC. Survivin and cyclin D1 might play synergetic roles in lung cancer cell' s karyokinesismitosis and they can be identified as potential therapeutic target in NSCLC.

Expression of cyclin D1, PCNA, RB and p16 associated with G1 cell-cycle in DMBA induced rat ovarian carcinogenesis / 대한산부인과학회잡지

OBJECTIVE: Ovarian cancer is common a gynecologic malignancy and leading cause of death in women being diagnosed with advanced disease. This study was undertaken to investigate the roles of the proteins related to G1 cell cycle in ovarian carcinogenesis. METHODS: The expression of cyclin Dl, p16, RB and PCNA in DMBA (7, 12-dimethylbenzanthracene)-induced ovarian cancer of rats was analyzed by immuno-histochemistry and Western blot. RESULTS: 1. Twenty-nine tumors were induced in 32 ovaries from 16 rats (90.6%) in the experimental group. The average weight of tumor was 3.35+/-0.73 gm and the average size was 1.84+/-0.17 cm in greatest dimension. The histologic types were adenocarcinomas (n=20), squamous cell carcinomas (n=3), sarcoma (n=4) and combined types (n=3). 2. With respect to the cyclin D1 and PCNA labelling index, ovarian cancers showed significantly higher index than normal ovarian surface epithelium. There were no differences among the cancer types. In Western blot analysis, the expression of cyclin Dl in ovarian cancers was higher than that in normal ovarian surface epithelium. 3. With respect to the p16 and RB labelling index, ovarian cancers showed significantly lower index than normal ovarian surface epithelium. There were no differences among the cancer types. In Western blot analysis, the expression of cyclin Dl in ovarian cancers were lower than that in normal ovarian surface epithelium. 4. Positive correlation was shown among cyclin D1, PCNA. RB was negatively correlated with cyclin D1, PCNA. The p16 had no correlation with cyclin D1, PCNA. CONCLUSION: These results suggest that the deregulation of cyclin Dl, p16, RB and PCNA occur in DMBA induced rat ovarian carcinogenesis and result in tumor progression. Further studies are needed to investigate the role and function of cyclin Dl-p16-RB pathway in human ovarian cancer with this animal model.

Expression of Cyclin D1 and CDK4 in DMBA-Induced Rat Ovarian Cancer / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Ovarian cancer is a common gynecologic malignancy and the leading cause of death in women. It is typically not diagnosed until it has reached the advanced stages. We performed this study to investigate the roles of the proteins related to the G1 cell cycle in ovarian carcinogenesis. MATERIALS AND METHODS: Immunohistochemistry and Western blot were used to analyse the expression of cyclin Dl and CDK4 in 7, 12-dimethylbenzanthracene- induced ovarian cancer in rats. RESULTS: The Cyclin D1 and CDK4 labelling index was significantly higher in the ovarian cancers than in the normal ovarian surface epithelium of rats. There was no difference among the cancer types. In Western blot analyses, the expression of cyclin Dl and CDK4 in the ovarian cancers was higher than that in the normal ovarian surface epithelium. A positive correlation was observed between the expressions of the CDK4 and Cyclin D1. CONCLUSION: The upregulation of cyclin Dl and CDK4 that occurs in DMBA-induced rat ovarian carcinogenesis is likely to be associated with tumor progression. Further studies are needed to investigate the role and function of cyclin Dl and CDK4 in human ovarian cancer.

Association Between Infection of Human Papillomavirus and Decreased Expression of Cyclin DI in Cervical Neoplasia / 대한부인종양콜포스코피학회잡지

Cyclin Dl, one of Gl cyclin gene subfamily, and human papillomaviruses (HPV) oncoprotein E7 have a homology in binding sites for the retinoblastoma tumor-suppressor protein. In order to evaluate the role of cyclin Dl in human cervical carcinogenesis, the level of its expression was measured and compared to HPV infection. In these studies, 38 normal control cases, 22 carcinoma in situ (CIN) cases, and 16 invasive cervical carcinomas were analyzed by immunocytochemistry and polymerase chain reactions for the detection of expression of cyclin Dl and infection of HPV type 16 and 18, respectively. The cyclin Dl expression was significantly lower in CIN and invasive carcinoma than normal control group regardless of HPV infection (p=0.026). The decreased expression of cyclin Dl in normal control group was not related with HPV infection. However, the levels of expression of cyclin Dl in CIN and invasive carcinoma were correlated with HPV 16and 18 (p 0.026). The expression of cyclin E was not changed in HPV 16 and 18 infected cases. These data provide the evidence that cyclin Dl expression in the lesions of cervical tumor is decreased and it is related with HPU infection.