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Objective:To study the expression and clinical significance of microRNA-574-3p (miR-574-3p) in colon cancer.Methods:A total of 106 colon cancer patients who were admitted to the First Hospital of Qinhuangdao and Shijiazhuang Hospital of Traditional Chinese Medicine from June 2012 to June 2015 were selected as the research objects. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of miR-574-3p in colon cancer tissues and normal adjacent tissues. The relationship between the expression of miR-574-3p and the clinicopathological characteristics and prognosis of patients with colon cancer was analyzed. Immunohistochemical staining was used to detect the relationship between the expression of miR-574-3p and the expression of CyclinA2 or E-cadherin.Results:Compared with normal tissues adjacent to cancer, the expression level of miR-574-3p in 106 cases of colon cancer was significantly lower ( P<0.01). The decreased expression of miR-574-3p was related to tumor diameter, Dukes stage, histological grade and lymph node metastasis (all P<0.05), but not to age and tumor location (all P>0.05). The patients with low expression of miR-574-3p, high Dukes stage and histological grade, and lymph node metastasis had poor survival (all P<0.05). The 5-year overall survival rate of patients with decreased miR-574-3p expression in cancer tissue was significantly lower than that of patients without decreased miR-574-3p expression ( P=0.007 6). Compared with patients with no decreased miR-574-3p expression, patients with decreased miR-574-3p expression had higher CyclinA2 protein integrated optical density (IOD) value and lower E-cadherin protein IOD value in colon cancer tissues (all P<0.05). Conclusions:The decreased expression of miR-574-3p is related to the poor prognosis of colon cancer patients, which may affect tumor recurrence and metastasis by regulating the expression of CyclinA2 and E-cadherin proteins.
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Objective:To investigate the diagnostic values of detections of human papillomavirus (HPV) DNA combined with peripheral blood cyclin A mRNA and cyclin-dependent kinase 2 (CDK2) mRNA for cervical squamous cell carcinoma.Methods:Eighty patients with cervical squamous cell carcinoma treated in Jiangyin Hospital of Traditional Chinese Medicine from January 2018 to October 2021 were selected as the research objects. Eighty patients with benign cervical lesions such as cervicitis treated in the same period were selected as the control. The levels of HPV-DNA in paraffin-embedded tissues of cervical squamous cell carcinoma were detected by gene chip, and the mRNA expression levels of cyclin A and CDK2 in peripheral blood monocytes were detected by reverse transcription polymerase chain reaction. The related factors of cervical squamous cell carcinoma were analyzed by multivariate logistic regression. Taking the results of pathological biopsy as the gold standard, the diagnostic efficacy of HPV-DNA, peripheral blood cyclin A mRNA and CDK2 mRNA single and combined detection for cervical squamous cell carcinoma were determined by receiver operating characteristic (ROC) curve.Results:The positive rate of HPV-DNA in patients with cervical squamous cell carcinoma was higher than that in the control group [75.00% (60/80) vs. 13.75% (11/80), P < 0.05]; the relative expressions of cyclin A mRNA and CDK2 mRNA in patients with cervical squamous cell carcinoma were 0.26±0.08 and 1.49±0.07, respectively, which were higher than those in the control group (0.11±0.03 and 1.14±0.06), and the differences were statistically significant (both P < 0.05). Multivariate logistic regression analysis showed that the number of induced abortions > 1 ( OR = 3.093, 95% CI 1.386-6.899, P = 0.021), the age of first birth ≤18 years old ( OR = 3.684, 95% CI 1.651-8.219, P = 0.013), the positive HPV-DNA ( OR = 4.125, 95% CI 1.849-9.202, P = 0.001), the increased relative expression of cyclin A mRNA in peripheral blood ( OR = 3.800, 95% CI 1.703-8.478, P = 0.006) and the increased relative expression of CDK2 mRNA in peripheral blood ( OR = 4.821, 95% CI 2.161-10.756, P = 0.008) were risk factors for the occurrence of cervical squamous cell carcinoma. ROC curve analysis showed that the area under the curve (AUC) of HPV-DNA, peripheral blood cyclin A mRNA and CDK2 mRNA in single and combined diagnosis of cervical squamous cell carcinoma were 0.769 (95% CI 0.700-0.838), 0.756 (95% CI 0.688-0.823), 0.755 (95% CI 0.689-0.820) and 0.827 (95% CI 0.766-0.888), respectively. Conclusions:HPV-DNA and the levels of cyclin A mRNA and CDK2 mRNA in peripheral blood can be used to assist in the diagnosis of cervical squamous cell carcinoma, and the combination of the three has high diagnostic efficiency.
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Objective: The present study delineates the generation of mutant peptide library from a known anticancer peptide, p21 and in silico evaluation for their affinity towards cyclin. A substrate binding groove. Methods: Mutant peptide library was created based on their AntiCP score and was docked with cyclin A using ClusPro2.0 web server. The docked structures were further simulated into an aqueous environment using Gromacs 4.5.6. Visualization was performed using PyMol software and interaction analysis was done using Discovery Studio Visualizer 4.1 Client and LigPlot plus tool. Results: A total of 57 mutant peptides were generated; out of which only 3 namely, K3C (Lys3Cys), K3F (Lys3Phe), and K3W (Lys3Trp) had a greater affinity for cyclin A than WILD p21 peptide (HSKRRLIFS). Molecular dynamic simulation studies showed that the peptides remained docked into the substrate binding groove throughout the run. Among all the peptides, K3C showed a significantly higher negative binding energy with cyclin A as compared to WILD. Conclusion: The overall results suggested that K3C mutant peptide had ~30 % higher affinity towards cyclin A and thus, could further be explored for its anticancer potential. The study also provides an insight into the crucial interactions governing the recognition of substrate binding groove of cyclin A for the development of novel peptide-based anticancer therapeutics.
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Objective To investigate the expression and clinical significance of DNMT1 and CCNA1 in different grades of cervical lesions. Methods Cervical tissues were selected from normal cervical(NC),low-grade cervical intraepithelial lesion(LSIL),high-grade cervical intraepithelial lesion(HSIL)and cervical squa-mous cell carcinoma(SCC)from each thirty patients.The expression of DNMT1 and CCNA1 mRNA and protein was examined by Western Blot analysis and qRT-PCR in cervical tissues. Results The expression of DNMT1 mRNA and protein in LSIL,HSIL and SCC was higher than in NC(F=117.93,P<0.05;F=61.24,P<0.05). Expression of DNMT1 mRNA and protein was increased steadily according to severity of cervical lesions(χ2trend=26.25,P<0.05;χ2trend=26.60,P<0.05).The expression of CCNA1 mRNA and protein in HSIL and SCC was lower than that in NC and LSIL(F = 77.04,P < 0.05;F = 57.15,P < 0.05). Expression of CCNA1 mRNA and protein was decreased steadily according to severity of cervical lesions(χ2trend=64.19,P<0.05;χ2trend=60.24,P<0.05). There was a negative correlation between expression of DNMT1 and CCNA1 protein in LSIL,HSIL,SCC (r=-0.75,-0.56,P<0.05). Conclusion In DNMT1 mRNA there is high expression of protein and in CCNA1 mRNA there is low expression of protein,but both may be related to the occurrence and development of cervical cancer.
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ABSTRACT:Objective To investigate the mechanism and induction of T type calcium channel on neural stem cells after brain injury .Methods Adult mice brain injury model was established and divided into control ,sham operation ,surgery ,and surgery+mebefradil groups .Neural stem cells were separated from the subventricular zone (SVZ) and identified .Moreover ,we analyzed the results using MTT and neural stem cell sphere counting after adding different doses of mibefradil in culture medium ,respectively .Then we calculated the half maximal inhibitory concentration (IC50) of mibefradil on neural stem cells .Finally ,we analyzed the expression of Cav3 .2 in SVZ and the protein expressions of Cav3 .2 ,Cyclin A and caspase‐3 in neural stem cells by Western blot .Results In vivo , neural stem cell proliferation was increased in surgery group compared with that in control and sham‐operation groups .The proliferation of neural stem cells in surgery + mibefradil groups was significantly decreased compared with that in surgery group after mibefradil‐induced inhibition of T type calcium channel protein . In vitro , the formation of neural stem cell sphere was significantly inhibited after adding mibefradil .The cell growth ratio was significantly decreased when the concentration of mibefradil was above 5μmol/L .A values in 5 μmol/L ,10 μmol/L and 20μmol/L groups were significantly lower than those in control group (P<0 .05) .IC50 was 8 .93μmol/L .The protein expressions of Cyclin A and Cav3 .2 were inhibited while that of caspase‐3 was increased after mibefradil treatment .Conclusion Neural stem cell proliferation was enhanced by activating T type calcium channel after brain injury .
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BACKGROUND:Cyclin A2 is a key regulator of cellcycle. Location and expression of cyclin A2 in neonatal mouse myocardium is not clear. OBJECTIVE:To observe the location and expression of cyclin A2 in neonatal mouse cardiomyocytes and its relationship with the exit of cardiomyocytes from cellcycle. METHODS:Neonatal mice were kil ed to take myocardial tissues at 0, 3, 7, 14 and 28 days after birth. Western blot were used to detect the expression of cyclin A2, proliferating cellnucleus antigen and Phospho-histone H3. Immunohistochemitry detection was used to detect the location of cyclin A2 and expression of proliferation cellnucleus antigen at different time after birth. RESULTS AND CONCLUSION:Western blot showed the decrease of cyclin A2 after birth til disappeared at day 4 (P=0.001). Cyclin A2 located mainly in the nucleus after birth and exported to the cytoplasm at day 14, and basical y disappeared at day 28. Proliferating cellnucleus antigen showed gradual y decreased tendency after birth. Mitosis specific marker, Phospho-histone H3, exhibited a gradual decrease after birth, which was consistent with cyclin A2 in expression intensity.
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Objective To explore the effect of Cyclin A on origin recognition complex-1 (ORC1) expression of rat vascular smooth cells (VSMCs).Methods Primary VSMCs of thoracic aorta in rats was obtained by the adherence method of tissue culture.The synchrony of cell was obtained by the method of double-thymidine block.In different cell cycles of VSMCs,the expression of ORC1 mRNA was determined by RT-PCR and the protein expression of ORC1 was observed by flow cytometry.Results Synchronized VSMCs were obtained and identified by the methods of double-thymidine block,colchicine treatment and serum starvation.Synchronized growth was monitored by flow cytometry.All the synchronized VSMC's distribution ration was (89.22±3.54) ％ at G0/G1 phase,(66.74 ±7.16)％ at G1/S phase,(63.24 ±4.06)％ at S phase and (51.64 ± 11.18)％ at G2/M phase and there was statistically significant difference compared with other phase(P ＜0.01).There was no significant effect of Cyclin A on ORC1 mRNA expression at a quiescent stage of VSMC.At G2/M phase peaked ORC1 was (52.133 3 ± 2.122 1)％,at G1/S phase was(10.916 7 ± 0.531 1)％,at S phase was (7.656 7 ± 0.412 4)％,and there was statistically significant difference (P ＜0.01).At G2/M phase ORC1 downed to the lowest point was (1.276 7 ± 0.161 7) ％,at G1/S phase was (13.371 0 ± 1.057 3)％,at S phase was (3.043 3 ± 0.538 0)％,and there was statistically significant difference (P ＜ 0.01),suggesting that Cyclin A might prevent ORC1 binding the chromatin of VSMCs.Conclusion Cyclin A may be an important regulative factor at the initiation of ORC1 in VSMCs.
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Dental epithelial and mesenchymal cells that form the teeth undergo dynamic changes in cell cycle during tooth development and morphogenesis. Although proliferation has been known as a key event during odontogenesis, the cell cycle phases and their relations with the complicated molecular mechanisms of tooth development are not fully understood yet. This study comparatively examined the expression patterns of Ki-67, cyclin A, and cyclin D1 during tooth development in the mouse incisor and molar in order to identify the cell-cycle characteristics during odontogenesis. We found that Ki-67 and cyclin A were expressed in the proliferating cells in the dental epithelial and mesenchymal tissues at the bud, cap and bell stages. Cycln D1 showed distinct expression in the incisor odontoblast region and the enamel knot, in which Ki-67 nor cyclin A was expressed. Our results provide specific information on the cell cycle phases during tooth development that may provide clues to relate them with the complex odontogenic mechanisms. Furthermore, we suggest that our findings enlightened the previous studies on the incisor odontoblasts and the enamel knot during tooth development.
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Animals , Mice , Cell Cycle , Cyclin A , Cyclin D1 , Cyclins , Dental Enamel , Incisor , Molar , Morphogenesis , Odontoblasts , Odontogenesis , Polymethacrylic Acids , ToothABSTRACT
BACKGROUND: Actinic keratosis (AK) and bowen's disease (BD) are pre-cancerous diseases, and are regarded as an early squamous cell carcinoma (SCC). AK and BD can be progressed into SCC. In this process, tumor suppressor and cell proliferative proteins may play important roles. OBJECTIVE: To investigate the differences of expression patterns of the immunohistochemical (IHC) staining and useful markers for differential diagnosis in AK, BD and SCC. METHODS: Biopsy had proven 17 cases of AK, 20 cases of BD and 17 cases of SCC, which were all selected. IHC staining for Ki-67 and cyclin-A, as cell proliferative markers, p53 and p16 as tumor suppressor markers, were performed. Labeling index (LI) and distribution pattern of IHC expressions were measured. RESULTS: LI of Ki-67 in AK, BD and SCC were 30.6%, 60.2% and 54.8%, respectively. LI of cyclin-A in AK, BD and SCC were 9.2%, 24.4% and 24.1%, respectively. LI of p53 in AK, BD and SCC were 20.7%, 37.9%, and 39.9%, respectively. LI of p16 in AK, BD and SCC were 10.6%, 38.3% and 39.9%, respectively. Lower 1/3 was the most frequent distribution pattern in AK in all IHC stains, full thickness lower 2/3 were the most frequent distribution pattern in BD and SCC in all IHC stains. CONCLUSION: LI and distribution pattern of Ki-67, cyclin-A, and p16, as well as the distribution pattern of p53 may be useful markers to differentiate AK from BD and SCC. Higher degree and full-thickness distribution pattern of IHC expressions in all stains may be helpful in the diagnosis of BD, rather than AK.
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Actins , Biopsy , Bowen's Disease , Carcinoma, Squamous Cell , Coloring Agents , Cyclin A , Cyclins , Diagnosis, Differential , Keratosis, Actinic , ProteinsABSTRACT
BACKGROUND: Hypoxia inducible factor-1alpha(HIF-1alpha) is a transcription factor for various target genes that are involved in adapting cells to hypoxia. It promotes cell proliferation and survival via modulation of such cell cycle regulators such as cyclin A1 and cyclin B1 in response to hypoxia. This is associated with local failure of radiotherapy, which renders a poor prognosis for cervical carcinoma. METHODS: Using the tissue histologic sections and a tissue microarray of the archived biopsy and surgical specimens of uterine cervical carcinoma from 57 patients who were treated with radiation therapy alone, we performed immunohistochemical staining for HIF-1alpha and cyclin A1 and B1 to evaluate the correlations between the expressions of these proteins in tumors and the clinicopathologic parameters associated with the prognosis. RESULTS: The large tumor cell nests and invasive front margins of the tumors showed comparatively intense immunoreactivity of HIF-1alpha. There was no significant correlation between the HIF-1alpha, cyclin A1 and cyclin B1 expressions and the clinicopathologic factors. CONCLUSIONS: The HIF-1alpha expression showed marked intra-tumoral heterogeneity. The HIF-1alpha expression is neither a powerful predictor of resistance to radiotherapy nor is it a poor prognostic marker in cervical carcinoma patients who are treated with radiotherapy. The expressions of cyclin A1 and cyclin B1 are neither independently associated with the response of radiation therapy nor are they associated with the prognostic parameters of uterine cervical carcinoma.
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Humans , Hypoxia , Biopsy , Cell Cycle , Cell Proliferation , Cyclin A , Cyclin A1 , Cyclin B , Cyclin B1 , Cyclins , Hypoxia-Inducible Factor 1, alpha Subunit , Population Characteristics , Prognosis , Proteins , Transcription Factors , Uterine Cervical NeoplasmsABSTRACT
Besides its function as a pathogenicity determinant, the Tombusvirus P19 also serves as a suppressor of RNA interference (RNAi) by sequestering intracellular small RNAs such as the small interfering RNAs (siRNAs) and microRNAs (miRNAs). However, the effect of P19 on mammalian cells has not been evaluated before. A human embryonic kidney 293 cell line that stably expressed p19 (HEK293-p19) was generated. Flow cytometric analysis revealed that over-expression of P19 caused a significant accumulation of G2/M phase cells. Cell proliferation assays demonstrated a reduced DNA replication and cell growth in HEK293-p19 cells. Moreover, p19 altered the expression profiles of a number of cell cycle regulators in HEK293 cells, such as upregulafion of cyclin A1, CDK2, CDK4, CDK6, p18, cyclin D2, p19INK4d and E2F1, and downregulation of p15, cyclin A2, cyclin B1 and cyclin E1. Thus, the data strongly indicate that p19 might influence multiple G2/M regulators to cause G2/M arrest.
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OBJECTIVE: This study was carried out to investigate the relationship between DNA ploidy, S-phase fraction (SPF), expression of cyclin A and clinical prognostic factors including stage, grade, CA-125 and residual tumor size in epithelial ovarian cancer, and to evaluate the association between DNA ploidy, SPF, expression of cyclin A and 3-year survival. METHODS: Study group consisted of 31 cases of epithelial ovarian cancer, 10 of borderline ovarian tumor and 5 of benign ovarian tumor diagnosed at the department of Obstet. and Gynecol. in Yonsei University College of Medicine, Seoul, Korea from Feb. 2000 to Jan. 2003. All patients underwent staging-laparotomy and postoperative chemotherapy. The level of CA-125 was assessed after 6th postoperative chemotherapy with cut-off value of 35 U/mL. DNA ploidy and SPF were evaluated by flow-cytometry of fresh ovarian tissue obtained at the operative field. The expression of cyclin A was evaluated by immuno-histochemical stain. Expression of 5% was considered as positive. Statistical analysis was done by two-sample t-test, chi-square test, and Kaplan-Meier survival curve using SPSS ver 11.0 software. RESULTS: In 46 ovarian tumors aneuploidy, SPF and expression of cyclin A were significantly higher in epithelial ovarian cancer as compared with benign and borderline tumors (p=0.004, 0.001, 0.001, respectively). Number of aneuploidy, SPF and expression of cyclin A were significantly higher in patients with higher grade, more advanced stage, higher level of CA-125 (more than 35 U/mL) and more than 2 cm of residual tumor size (p=0.004, 0.009, 0.05, 0.002 in aneuploidy; p=0.06, 0.01, 0.04, 0.007 in SPF; p=0.03, 0.004, 0.06, 0.02 in cyclin A). Aneuploidy and expressions of more than 10% of SPF and cyclin A were also associated with poorer overall survival (p=0.02, 0.02, <0.0001, respectively). Significantly positive correlations were observed among these factors. CONCLUSION: Number of aneuploidy, percentage of SPF and expression of cyclin A were higher in more advanced stage, higher grade, higher CA-125 and more than 2 cm of residual tumor size and associated with poorer overall survival. Thus DNA flow-cytometry and estimation of expression of cyclin A may provide major information about prognosis of disease in epithelial ovarian cancer patients.
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Humans , Aneuploidy , Cyclin A , Cyclins , DNA , Drug Therapy , Korea , Neoplasm, Residual , Ovarian Neoplasms , Ploidies , Prognosis , SeoulABSTRACT
Methyl-beta-cyclodextrin, a cyclic oligosaccharide known for its interaction with the plasma membrane induces several events in cells including cell growth and anti-tumor activity. In this study, we have investigated the possible role of cyclooxygenase 2 (COX-2) in cell growth arrest induced by methyl-beta-cyclodextrin in Raw264.7 macrophage cells. Methyl-beta-cyclodextrin inhibited cell growth and arrested the cell cycle, and this cell cycle arrest reduced the population of cells in the S phase, and concomitantly reduced cyclin A and D expressions. Methyl-beta-cyclodextrin in a dose- and time-dependent manner, also induced COX-2 expression, prostaglandin E(2) (PGE(2)) synthesis, and COX-2 promoter activity. Pretreatment of cells with NS398, a COX-2 specific inhibitor completely blocked PGE(2) synthesis induced by methyl-beta-cyclodextrin, however inhibition on cell proliferation and cell cycle arrest was not effected, suggesting non-association of COX-2 in the cell cycle arrest. These results suggest that methyl-beta-cyclodextrin induced cell growth inhibition and cell cycle arrest in Raw264.7 cells may be mediated by cyclin A and D1 expression.
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Animals , Mice , Cell Cycle/drug effects , Cell Line , Cell Proliferation/drug effects , Dinoprostone/metabolism , Dose-Response Relationship, Drug , Isoenzymes/genetics , Macrophages/cytology , Prostaglandin-Endoperoxide Synthases/genetics , beta-Cyclodextrins/pharmacologyABSTRACT
Objective To investigate the possible roles of Cyclin A and P21 in the pathogenesis and development of endometriosis.Methods Twenty-five specimens of eutopic endometrium and twenty-eight specimens of ectopic endometrial tissues were obtained from 28 patients with endometriosis (observation group).Thirty-five specimens of endometrium were also obtained from women with other benign diseases (control group).Cyclin A and P21 were detected by Western blotting,RT-PCR and immunohistochemistry.Results The expression level of cyclin A mRNA in ectopic endometrium were (1.24?0.10) at the proliferative phase and (1.33?0.21) at the secretory phase,significantly higher than that in the eutopic endometrium tissues (0.72?0.26,0.42?0.22),control group (0.68?0.09,0.35?0.06) (P0.05).The expression level of P21 mRNA in control group were (0.21?0.01) at the proliferative phase and (0.54?0.35) at the secretory phase,significantly higher than that in the eutopic endometrium tissues (0.09?0.06,0.28?0.02) and in the ectopic endometrium (0.06?0.01,0.13?0.00) (P0.05).In the ectopic endometrium they had negatively spearman correlation (r=-0.738,P
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Objective To investigate the effect of cyclin A antisense oligodeoxynucleotide (ASON) on mediating proliferation of K562 cell. Method After in vitro co-culture of cyclin A ASON with K562 cell, cyclin A protein expression levels were measured by flow cytometry. Cell growth was detected by trypan blue dye exclusion and colony-forming experiment. Results In cyclin A ASON group, cyclin A protein expression was significantly inhibited, compared to those in sense oligodeoxynucleotide (SON) group and blank group. Moreover, when the ASON concentration increased, the proliferation ratio of K562 cells and the CFU- K562 were significantly inhibited. Conclusion Cyclin A ASON can specifically inhibit cyclin A protein expression as well as inhibit the K562 cell proliferation and can lead to leukemic cell apoptosis. The effect of cyclin A ASON is concentration dependent.
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AIM: To investigate the influence of matrine on cyclin E and cyclin A in U937 cell strain. METHODS: The inhibitory effect of matrine on proliferation in U937 cell strain was observed by MTT.The distribution of cell cycle was measured by flow cytometry.Immunocytochemical method and Western blot were used to detect the protein expression of cyclin E and cyclin A. RESULTS: After being treated with matrine,the proliferation of U937 cell strain was inhibited significantly,the inhibiting effect conformed to time and dose-dependence,cells were arrested in S-phase.The expression of cyclin E and cyclin A was down-regulated significantly in a dose-dependence manner. CONCLUSION: Matrine could reduce the expression of cyclin E and cyclin A and inhibit the proliferation in U937 cell strain.