目的 探討乳腺癌中CyclinD1基因擴(kuò)增和其蛋白表達(dá)的相關(guān)性及其臨床意義。方法 應(yīng)用半定量PCR和免疫組化技術(shù)，檢測和分析了乳腺癌及其癌旁組織、乳腺良性組織及正常乳腺標(biāo)本中CyclinD1基因的擴(kuò)增和CyclinD1蛋白的表達(dá)情況。結(jié)果 62例乳腺癌中，CyclinD1基因擴(kuò)增占22.6%(14/62)，蛋白過度表達(dá)占48.4%(30/62)，二者有一定相關(guān)性，而其它各種乳腺組織的CyclinD1基因擴(kuò)增及蛋白過度表達(dá)與之相比差異有顯著性意義(Ｐ＜0.05)。CyclinD1基因擴(kuò)增及蛋白過度表達(dá)與乳腺癌組織學(xué)分級呈明顯正相關(guān)，且CyclinD1蛋白過度表達(dá)還與雌孕激素受體狀況呈正相關(guān)。結(jié)論 乳腺癌中CyclinD1基因擴(kuò)增程度和其蛋白過度表達(dá)雖有一定相關(guān)性，但并不完全一致，說明還存在著其它導(dǎo)致CyclinD1蛋白過度表達(dá)的機(jī)理，CyclinD1蛋白的表達(dá)可能還受雌激素的調(diào)控。對乳腺癌中CyclinD1基因及其蛋白表達(dá)的檢測有一定臨床意義。
【摘要】 目的 觀察晚期糖基化終產(chǎn)物（advanced glycosylation end prodrcts，AGE）對人結(jié)腸癌細(xì)胞株SW-480增殖的影響，并探討其可能機(jī)制。 方法 不同濃度AGE干預(yù)SW-480細(xì)胞，噻唑藍(lán)（MTT）法比較各組細(xì)胞活力，流式細(xì)胞術(shù)觀察AGE對SW-480細(xì)胞周期的影響，蛋白質(zhì)印跡法觀察AGE對SW-480細(xì)胞CyclinD1表達(dá)的影響，端粒重復(fù)序列擴(kuò)增法（telomeric repeat amplification protocol，TRAP）銀染法觀察AGE對SW-480細(xì)胞端粒酶活性的影響。MTT測細(xì)胞活力的檢測設(shè)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(shè)置100 μg/mL BSA組和100 μg/mL AGE組。 結(jié)果 MTT結(jié)果示AGE促進(jìn)SW-480細(xì)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(xì)胞G0/G1期所占百分比分別為56.02%±0.58%、51.93%±1.01%，差異有統(tǒng)計學(xué)意義（Plt;0.05）。蛋白質(zhì)印跡法示100 μg/mL AGE組72 h后CyclinD1的表達(dá)較100 μg/mL BSA組增加，差異有統(tǒng)計學(xué)意義（Plt;0.05）。TRAP銀染法檢測示100 μg/mL AGE干預(yù)SW-480細(xì)胞72 h后可以增加端粒酶活性（Plt;0.05）?！〗Y(jié)論 AGE可促進(jìn)人結(jié)腸癌細(xì)胞SW-480生長，呈劑量依賴性。其作用機(jī)制可能與AGE上調(diào)CyclinD1的表達(dá)加速G1/S期轉(zhuǎn)換及增加端粒酶活性有關(guān)?！続bstract】 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 觀察CyclinD1在食管鱗狀細(xì)胞癌和癌周正常食管組織的表達(dá)，從量化角度闡明其在食管鱗狀細(xì)胞癌組織中表達(dá)的病理學(xué)意義及其與CyclinE表達(dá)的關(guān)系。 方法 用免疫組化MaxVision法檢測CyclinD1、CyclinE在食管鱗狀細(xì)胞癌和癌周正常食管組織的表達(dá)，Image-pro plus圖像分析軟件對其表達(dá)強(qiáng)度進(jìn)行定量分析，并用表達(dá)的陽性單位（positive unit，PU）反映其表達(dá)強(qiáng)度。 結(jié)果 CyclinD1蛋白在食管鱗狀細(xì)胞癌癌細(xì)胞核中表達(dá)的PU值高于正常食管組織（Plt;0.001）；CyclinD1蛋白表達(dá)的PU值與食管鱗狀細(xì)胞癌分化程度有關(guān)（Plt;0.001）；浸潤漿膜層組的CyclinD1蛋白表達(dá)的PU值高于無漿膜層浸潤組（P=0.037）；伴淋巴結(jié)轉(zhuǎn)移組的CyclinD1蛋白表達(dá)的PU值高于無淋巴結(jié)轉(zhuǎn)移組（P=0.012）；CyclinD1與CyclinE蛋白在食管鱗狀細(xì)胞癌中表達(dá)呈正相關(guān)（r=0.650，Plt;0.01）。 結(jié)論 CyclinD1在食管鱗狀細(xì)胞癌中表達(dá)上調(diào)，且與生物學(xué)特性相關(guān)，可作為判斷腫瘤患者預(yù)后不良的標(biāo)志之一；CyclinD1和CyclinE可能在食管癌癌變過程中起協(xié)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:觀察CyclinD1,P16在皮膚鱗狀細(xì)胞癌組織中的表達(dá),探討CyclinD1,P16在皮膚鱗狀細(xì)胞癌發(fā)生發(fā)展中的作用。方法:用免疫組織化學(xué)技術(shù)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(dá)。結(jié)果:CyclinD1在皮膚鱗癌組織中表達(dá)高于皮膚正常對照組,兩者差異有顯著統(tǒng)計學(xué)意義(Plt;0.01); P16在皮膚鱗癌組織中表達(dá)低于皮膚正常對照組,但兩者差異無顯著統(tǒng)計學(xué)意義(Pgt;0.05);CyclinD1,P16在皮膚鱗癌中表達(dá)呈正相關(guān)性(r=0.782,Plt;0.01)。結(jié)論:CyclinD1在皮膚鱗狀細(xì)胞癌組織中陽性表達(dá)上調(diào),P16在皮膚鱗狀細(xì)胞癌組織中陽性表達(dá)下調(diào),CyclinD1,P16在皮膚鱗癌中表達(dá)呈正相關(guān)。