ObjectiveTo investigate the expression of Cyclin D1 in stage T2-3N0M0 esophageal squamous cell carcinoma (ESCC) and its relationship with patient prognosis. MethodsExpression of Cyclin D1 in 227 ESCC specimens at stage T2-3N0M0 was detected by immunohistochemistry (IHC). Receiver operating characteristic (ROC) curve analysis was performed to select the cut-off score for high or low IHC reactivity. Correlations between Cyclin D1 expression and clinicopathological features as well as prognosis of ESCC patients were determined. ResultsAmong the 227 patients, 90 (39.6%) patients had low expression of Cyclin D1, and 137 (60.4%) patients had high expression of Cyclin D1.No significant correlation was observed between Cyclin D1 expression and patient gender (P=0.298), age(P=0.525), tumor location (P=0.570), tumor length (P=0.056), tumor grade (P=0.713), and depth of invasion (P=0.557). There was significant difference in prognosis between low-expression Cyclin D1 group and high-expression Cyclin D1 group (3-year survival rate:51.1% vs. 43.8%, 5-year survival rate:43.3% vs. 30.7%, P=0.047). Cox proportional-hazards regression model analysis showed that Cyclin D1 was not an independent prognostic factor for ESCC patients at stage T2-3N0M0 (relative risk=1.378, 95% CI:0.990-1.919, P=0.057). ConclusionOver expression of Cyclin D1 may be an adverse prognostic factor of ESCC patients at stage T2-3N0M0.
ObjectiveTo detect the expression of cyclin D1 in different kinds of clinical molecular subtypes of breast cancer tissue, and to analyze the relationships of the expression to clinicopathologic characteristics and prognosis. MethodsThe expression of cyclin D1 was detected in 175 cases of different kinds of clinical molecular subtypes of breast cancer by immunohistochemical method.The expression was compared among different kinds of clinical molecular subtypes of breast cancer by chi-square test.The correlations with clinicopathologic characteristics were analyzed by Spearman rank correlation.The impact of different expression of cyclin D1 on the prognosis was analyzed by the Kaplan-Meier survival curve. Results①In the positive expression group of cyclin D1 in the breast cancer, Luminal A type was the highest proportion (P < 0.05);but in the negative expression group of cyclin D1 in the breast cancer, Luminal B type was the highest proportion (P < 0.05).②The expression of cyclin D1 had no correlation with clinical stage, histological grade or PR (P > 0.05), was positively correlated with number of lymph node metastasis (rs=0.197, P < 0.01) or ER (rs=0.139, P < 0.05), was negatively correlated with Her-2(rs=-0.131, P < 0.05).③The positive expression of cyclin D1 was stronger, the tumor free survival time was longer (P < 0.05). ConclusionThis study shows that there is a correlation between the positive expression of cyclin D1 and ER, Her-2, or number of lymph node metastasis, and its positive expression prompts a relatively good prognosis, can be used as the prognosis indicator for breast cancer.
Objective To investigate the clinical significance of the amplification of CyclinD1 gene and overexpression of its protein product in breast cancer. Methods Semi-quantitative PCR and immunohistochemical techniques were used to examine the amplification of CyclinD1 gene and overexpression of its protein product in breast cancer, mammary tissues adjacent to the tumor, mammary benign disease and normal mammary tissues. Results The results showed that amplification of CyclinD1 gene was observed in 14 out of 62 (22.6%) breast cancer, the overexpression of its protein product was 48.4% (30/62). Compared with other mammary tissues, these findings were significant. Amplification of CyclinD1 gene and overexpression of its protein were positively associated with histological grade, and there was a b correlation between CyclinD1 protein overexpression and estrogen receptor (progestogen receptor). Conclusion The degree of CyclinD1 protein overexpression is not quite the same as the frequency of the amplification of CyclinD1 gene. It suggests that the overexpresstion of CyclinD1 protein is also attributed to some other mechanisms. Estrogen might stimulate the expression of CyclinD1. The detection of CyclinD1 gene and its protein in breast cancer has some clinical significances.
【Abstract】ObjectiveTo investigate the inhibitory effects of somatostatin analogue (SSTA) on the colonic carcinoma cell growth in vitro and in vivo and its possible mechanism. MethodsThe somatostatin receptor type Ⅱ (SSTR2) mRNA of colonic carcinoma cell line HCT116 was detected by using RTPCR and hybridization in situ. The effects of octreotide (Oct) or NC-8-12 (specific agonist of SSTR2 ) on the proliferation of HCT116 was measured with MTT after HCT116 stimulated by insulin or epidermal growth factor (EGF) and incubated with Oct or NC-8-12 simultaneously for 24 hours. The expression of cyclin D1 was detected with flow cytometry. The HCT116 were implanted in nude mice subcutaneously and treated with Oct or NC-8-12. The tumor volume and tumor weight were measured according to schedule. Results①SSTR2 mRNA was detected in HCT116 and the tumor implanted in nude mice; ②Insulin and EGF increased the proliferation of HCT116 significantly, and this proliferation could be inhibited by Oct and NC-8-12 partially; ③Insulin increased the Cyclin D1 expression of HCT116, its level decreased slightly when treated with Oct or NC-8-12 but not significantly (Pgt;0.05); ④The weight and volume of implanted tumor in nude mice treated with Oct or NC-8-12 showed no significant difference compared with the control group (Pgt;0.05). ConclusionBoth Oct and NC-8-12 could inhibit the proliferation of colonic carcinoma cell line HCT116 in vitro, which indicated that SSTR2 may mediated the inhibition. Oct and NC-8-12 have no effect on the growth of implanted HCT116 in nude mice in this experiment.
目的:觀察CyclinD1,P16在皮膚鱗狀細胞癌組織中的表達,探討CyclinD1,P16在皮膚鱗狀細胞癌發生發展中的作用。方法:用免疫組織化學技術檢測40例皮膚鱗癌石蠟包埋組織和10例正常皮膚組織的CyclinD1,P16表達。結果:CyclinD1在皮膚鱗癌組織中表達高于皮膚正常對照組,兩者差異有顯著統計學意義(Plt;0.01); P16在皮膚鱗癌組織中表達低于皮膚正常對照組,但兩者差異無顯著統計學意義(Pgt;0.05);CyclinD1,P16在皮膚鱗癌中表達呈正相關性(r=0.782,Plt;0.01)。結論:CyclinD1在皮膚鱗狀細胞癌組織中陽性表達上調,P16在皮膚鱗狀細胞癌組織中陽性表達下調,CyclinD1,P16在皮膚鱗癌中表達呈正相關。
ObjectiveTo investigate the clinical electrophysiological characteristics of Cyclin-dependent kinase-like 5 gene induced developmental epileptic encephalopathy (CDKL5-DEE). MethodsThe clinical data and series of video EEGs of children with CDKL5-associated developmental epileptic encephalopathy (CDKL5-DEE) who were admitted to the Children’s Medical Center of Peking University First Hospital from June 2016 to May 2024 were retrospectively analyzed. Results A total of 16 patients with CDKL5-DEE were enrolled, including 13 females and 3 males. All patients had de novo variants of CDKL5 gene, including 6 cases of missense variants, 5 cases of frameshift variants, 4 cases of nonsense variants, and 1 case of large fragment deletion. The age of onset was 8 days (d) after birth ~1 year (y) and 10 months (m), and the median age was (85.94±95.76) days. Types of seizures at onset: 4 cases of tonic seizures [age of onset 10~52 days, median age (25.5±15.84) days]; There were 5 cases of focal seizures [age of onset 8 d~8 m, median age (77.76±85.97) d]. There were 4 cases of epileptic spasmodic seizures [age of onset 3 m~1 y 10 m, median age (6.25±3.49) m]; There were 2 cases of bilateral tonic-clonic seizures [age of onset 30~40 days, median age (35.00±5.00) days]; focal concurrent epileptic spasm seizures 1 case (age of onset 2 m). A total of 59 VEEG sessions were performed in the pediatric EEG room of Peking University First Hospital for 4 hours. All the results were abnormal, including 26 normal background, 25 slow rhythm difference with background, and 8 no background. The interictal was 16 posterior or focal discharges, 19 multifocal discharges, 17 generalized or accompanied by focal/multifocal discharges, and 7 hypsarrhythmia; The ictal was 33 epileptic seizures, 6 myoclonic seizures, 5 focal seizures, 2 tonic-clonic seizures, 2 atypical absence seizures, 2 tonic seizures, 1 myoclonic sequential focal seizure, 1 focal sequential epileptic spasm, and 1 hypermotor-tonic-spasms. The background of patients within 6 months of age was normal, and the background abnormality increased significantly with age. generalized discharges are evident after 2 years of age between seizures. Conclusion CDKL5-DEE seizures have an early onset and are refractory to medications. Epileptic spasms are the most common type of seizure in every patient and long-lasting, with generalized seizures increasing markedly with age. EEG is characterized by a normal background within 6 months. With the increase of age, the background and interictal discharges have a tendency to deteriorate.
Objective To study the expression and its significance of cyclin D1,cyclin E and p27 protein in gastric carcinoma. Methods The expression of cyclin D1,cyclin E and p27 protein in 60 cases of gastric carcinoma were detected by immunostaining method. Results The positive rates of cyclin D1,cyclin E and p27 protein expression in gastric carcinoma were 41.7%（25/60）,38.3%（23/60）and 36.7%（22/60）,respectively.The expression of cyclin D1 was significantly correlated with Lauren type of tumor,the expression of cyclin E was significantly correlated with tumor differentiation,infiltration and stage,while the expression of p27 protein was significantly correlated with tumor infiltration,lymph nodes metastasis and stage. Further more, the expression of cyclin E was inversely correlated with the expression of p27. Conclusion Detection of cyclin D1,cyclin E and p27 expression in gastric carcinoma may be helpful in determining tumor progression.
Objective To explore effects of zinc on the contents of cycl in D2, cycl in-dependent kinase 4 (CDK4), and their DNA and total cellular protein in human umbil ical cord blood-drived mesenchymal stem cells (hUCBMSCs). Methods hUCBMSCs were isolated and cultured by density gradient centrifugation adherence method in vitro. At the serial subcultivation, the hUCBMSCs were randomly divided into 7 groups. In control group, hUCBMSCs were cultured with DMEM medium (containing 15%FBS). In treatment groups, hUCBMSCs were cultured with DMEM medium (containing 15%FBS plusZnSO4?7H2O). The final concentrations of zinc were 0.5, 1.5, 2.5, 3.5, 4.5, and 5.5 mg/L, respectively. The cellular surface antigens of CD29, CD34, CD44, and CD45 at the 3rd generation of hUCBMSCs were detected by flow cytometry. MTT assay was used to detect cell activity of the 3rd generation of hUCBMSCs. The optimum concentration of zinc was selected by the results of MTT as experimental group. The cell growth curves of experimental group and control group were drown by counting cell. The cell surface antigen, reproductive cycle, and DNA content were detected by flow cytometry motheds. The contents of cycl in D2 and CDK4 were detected by Western blot method. Results The positive expression rates of CD29 and CD44 were more than 70% in hUCBMSCs. The cell activity of 2.5 mg/L treatment group was superior to other treatment groups, as experimental group. At 7, 14, and 28 days, the contents of DNA, total cellular protein, cycl in D2, and CDK4 of hUCBMSCs were significantly higher in experimental group than those in control group (P lt; 0.01). The percentage of hUCBMSCs at S stage and prol iferation index in experimental group were also significantly higher than those in control group (P lt; 0.01). Conclusion Zinc (0.5-4.5 mg/L) has the promoting effect on the hUCBMSCs activity, and 2.5 mg/L is the optimal concentration. Zinc (2.5 mg/L) can accelerate the prol iferation and DNA reproduction of hUCBMSCs and increase the contents of cycl in D2 , CDK4, and cellular total protein.
【摘要】 目的 觀察CyclinD1在食管鱗狀細胞癌和癌周正常食管組織的表達，從量化角度闡明其在食管鱗狀細胞癌組織中表達的病理學意義及其與CyclinE表達的關系。 方法 用免疫組化MaxVision法檢測CyclinD1、CyclinE在食管鱗狀細胞癌和癌周正常食管組織的表達，Image-pro plus圖像分析軟件對其表達強度進行定量分析，并用表達的陽性單位（positive unit，PU）反映其表達強度。 結果 CyclinD1蛋白在食管鱗狀細胞癌癌細胞核中表達的PU值高于正常食管組織（Plt;0.001）；CyclinD1蛋白表達的PU值與食管鱗狀細胞癌分化程度有關（Plt;0.001）；浸潤漿膜層組的CyclinD1蛋白表達的PU值高于無漿膜層浸潤組（P=0.037）；伴淋巴結轉移組的CyclinD1蛋白表達的PU值高于無淋巴結轉移組（P=0.012）；CyclinD1與CyclinE蛋白在食管鱗狀細胞癌中表達呈正相關（r=0.650，Plt;0.01）。 結論 CyclinD1在食管鱗狀細胞癌中表達上調，且與生物學特性相關，可作為判斷腫瘤患者預后不良的標志之一；CyclinD1和CyclinE可能在食管癌癌變過程中起協同作用。/【Abstract】 Objective To observe the expression of CyclinD1 and its correlation with the expression of CyclinE in esophagus squamous carcinoma. Methods The expressions of CyclinD1 and CyclinE protein were detected by immunohistochemistry (MaxVision method) using the monoclonal antibody and their intensities were assessed quantitatively by Image-pro plus image analysis system. Results The monoclonal antibody was detected in specific cell nucleus. The positive unit (PU) value of CyclinD1 was larger in the esophagus squamous carcinoma than that in the normal esophagus tissue with a statistically significant difference (Plt;0.001). The expression of CyclinD1 was correlated with differentiation of esophagus squamous carcinoma (Plt;0.001) and infiltration degree of esophagus squamous carcinoma (P=0.037). The PU value of CyclinD1 of the nuclei was larger in esophagus squamous carcinoma with lymph node metastasis than those without lymph node metastasis (P=0.012). There was a positive correlation between CyclinD1 protein and CyclinE protein (r=0.650, Plt;0.01). Conclusion The positive expression of CyclinD1 is up-regulated in the esophagus squamous carcinoma. CyclinD1 protein relates to the clinicopathological characteristics and prognostic value in esophagus squamous carcinoma. CyclinD1 expression is bly associated with CyclinE expression in esophagus squamous carcinoma.
【摘要】 目的 觀察晚期糖基化終產物（advanced glycosylation end prodrcts，AGE）對人結腸癌細胞株SW-480增殖的影響，并探討其可能機制。 方法 不同濃度AGE干預SW-480細胞，噻唑藍（MTT）法比較各組細胞活力，流式細胞術觀察AGE對SW-480細胞周期的影響，蛋白質印跡法觀察AGE對SW-480細胞CyclinD1表達的影響，端粒重復序列擴增法（telomeric repeat amplification protocol，TRAP）銀染法觀察AGE對SW-480細胞端粒酶活性的影響。MTT測細胞活力的檢測設置空白對照組、100 μg/mL小牛血清白蛋白（bovine serum albumin，BSA）組及50、100、500 μg/mL AGE組，其余檢測只設置100 μg/mL BSA組和100 μg/mL AGE組。 結果 MTT結果示AGE促進SW-480細胞的增殖，且呈濃度依賴性。100 μg/mL BSA組與100 μg/mL AGE組72 h后的細胞G0/G1期所占百分比分別為56.02%±0.58%、51.93%±1.01%，差異有統計學意義（Plt;0.05）。蛋白質印跡法示100 μg/mL AGE組72 h后CyclinD1的表達較100 μg/mL BSA組增加，差異有統計學意義（Plt;0.05）。TRAP銀染法檢測示100 μg/mL AGE干預SW-480細胞72 h后可以增加端粒酶活性（Plt;0.05）。 結論 AGE可促進人結腸癌細胞SW-480生長，呈劑量依賴性。其作用機制可能與AGE上調CyclinD1的表達加速G1/S期轉換及增加端粒酶活性有關。【Abstract】 Objective To observe the effects of advanced glycosylation end products (AGE) on proliferation of SW-480 cells and study the possible mechanism. Methods Various concentrations of AGE were designed to have impact on SW-480 cells. Proliferation of SW-480 cells was assessed by thiazolyl blue tetrazolium bromide (MTT) assay; The impact of AGE on the cell cycle of SW-480 cells was analyzed by flow cytometry (FCM); the influence of AGE on expression of CyclinD1 was checked by Western blotting; and the impact of AGE on telomerase activity was examined by telomeric repeat amplification proctol (TRAP) sliver staining. For the MTT assay, blank control group, 100 μg/mL bovine serum albumin (BSA) group, 50, 100 and 500 μg/mL AGE groups were designed, while for other examinations, there were only 100 μg/mL BSA group and 100 μg/mL AGE group. Results MTT result showed that AGE increased the proliferation of SW-480 cells in a dose-dependent mode. The proportion of the cells at G0/G1 stage of the 100 μg/mL BSA group and the 100 μg/mL AGE experimental group were (56.02±0.58)% and (51.93±1.01)% respectively after 72 hours, with a significant difference (Plt;0.05); western blotting showed that the expression of CyclinD1 in the 100 μg/mL AGE group was significantly higher than that in the 100 μg/mL BSA group after 72 hours; TRAP silver staining demonstrated that telomerase activity increased significantly after treated with 100 μg/mL AGE for 72 hours. Conclusions AGE can promote the growth of SW-480 cells in a dose-dependent mode. Its mechanism is mainly by up-regulating the expression of CyclinD1 to shorten G0/G1 and increasing the telomerase activity significantly.