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.
【摘要】 目的 观察晚期糖基化终产物（advanced glycosylation end prodrcts，AGE）对人结肠癌细胞株SW-480增殖的影响，并探讨其可能机制。 方法 不同浓度AGE干预SW-480细胞，噻唑蓝（MTT）法比较各组细胞活力，流式细胞术观察AGE对SW-480细胞周期的影响，蛋白质印迹法观察AGE对SW-480细胞CyclinD1表达的影响，端粒重复序列扩增法（telomeric repeat amplification protocol，TRAP）银染法观察AGE对SW-480细胞端粒酶活性的影响。MTT测细胞活力的检测设置空白对照组、100 μg/mL小牛血清白蛋白（bovine serum albumin，BSA）组及50、100、500 μg/mL AGE组，其余检测只设置100 μg/mL BSA组和100 μg/mL AGE组。 结果 MTT结果示AGE促进SW-480细胞的增殖，且呈浓度依赖性。100 μg/mL BSA组与100 μg/mL AGE组72 h后的细胞G0/G1期所占百分比分别为56.02%±0.58%、51.93%±1.01%，差异有统计学意义（Plt;0.05）。蛋白质印迹法示100 μg/mL AGE组72 h后CyclinD1的表达较100 μg/mL BSA组增加，差异有统计学意义（Plt;0.05）。TRAP银染法检测示100 μg/mL AGE干预SW-480细胞72 h后可以增加端粒酶活性（Plt;0.05）。 结论 AGE可促进人结肠癌细胞SW-480生长，呈剂量依赖性。其作用机制可能与AGE上调CyclinD1的表达加速G1/S期转换及增加端粒酶活性有关。【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.
【摘要】 目的 观察CyclinD1在食管鳞状细胞癌和癌周正常食管组织的表达，从量化角度阐明其在食管鳞状细胞癌组织中表达的病理学意义及其与CyclinE表达的关系。 方法 用免疫组化MaxVision法检测CyclinD1、CyclinE在食管鳞状细胞癌和癌周正常食管组织的表达，Image-pro plus图像分析软件对其表达强度进行定量分析，并用表达的阳性单位（positive unit，PU）反映其表达强度。 结果 CyclinD1蛋白在食管鳞状细胞癌癌细胞核中表达的PU值高于正常食管组织（Plt;0.001）；CyclinD1蛋白表达的PU值与食管鳞状细胞癌分化程度有关（Plt;0.001）；浸润浆膜层组的CyclinD1蛋白表达的PU值高于无浆膜层浸润组（P=0.037）；伴淋巴结转移组的CyclinD1蛋白表达的PU值高于无淋巴结转移组（P=0.012）；CyclinD1与CyclinE蛋白在食管鳞状细胞癌中表达呈正相关（r=0.650，Plt;0.01）。 结论 CyclinD1在食管鳞状细胞癌中表达上调，且与生物学特性相关，可作为判断肿瘤患者预后不良的标志之一；CyclinD1和CyclinE可能在食管癌癌变过程中起协同作用。/【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.
目的:观察CyclinD1,P16在皮肤鳞状细胞癌组织中的表达,探讨CyclinD1,P16在皮肤鳞状细胞癌发生发展中的作用。方法:用免疫组织化学技术检测40例皮肤鳞癌石蜡包埋组织和10例正常皮肤组织的CyclinD1,P16表达。结果:CyclinD1在皮肤鳞癌组织中表达高于皮肤正常对照组,两者差异有显著统计学意义(Plt;0.01); P16在皮肤鳞癌组织中表达低于皮肤正常对照组,但两者差异无显著统计学意义(Pgt;0.05);CyclinD1,P16在皮肤鳞癌中表达呈正相关性(r=0.782,Plt;0.01)。结论:CyclinD1在皮肤鳞状细胞癌组织中阳性表达上调,P16在皮肤鳞状细胞癌组织中阳性表达下调,CyclinD1,P16在皮肤鳞癌中表达呈正相关。