目的研究胃癌增殖細(xì)胞核抗原(proliferating cell nuclear antigen，PCNA)表達(dá)與腹腔灌洗液端粒酶活性及腹膜轉(zhuǎn)移的相關(guān)性，并比較腹腔灌洗液中端粒酶活性和細(xì)胞學(xué)檢測游離癌細(xì)胞預(yù)測腹膜轉(zhuǎn)移的應(yīng)用價值。方法應(yīng)用免疫組化SP法檢測60例胃癌患者胃癌組織中PCNA表達(dá),PCRTRAPELISA法檢測腹腔灌洗液中端粒酶活性,同時行腹腔灌洗液脫落細(xì)胞學(xué)(peritoneal lavage cytology,PLC)檢測； 并分析其與相關(guān)臨床病理因素的關(guān)系。結(jié)果胃癌患者腹腔灌洗液中端粒酶活性的陽性率為41.7%； 與漿膜侵犯、組織學(xué)類型、浸潤深度、漿膜受累面積及腹膜轉(zhuǎn)移密切相關(guān)，并隨著浸潤深度及漿膜受累面積的增加而升高(P＜0.05)。PLC檢測陽性率為25.0%； 在伴肉眼可見腹膜轉(zhuǎn)移灶(P1～3)者明顯增高，也隨著浸潤深度及漿膜受累面積的增加而升高。兩種方法檢測的陽性率總體上差異無統(tǒng)計學(xué)意義，但在未分化型癌、pT4、伴肉眼可見腹膜轉(zhuǎn)移灶(P1～3)者端粒酶活性陽性率明顯高于PLC。PCNA增殖指數(shù)（PI）在腹腔灌洗液端粒酶活性表達(dá)陽性者明顯高于表達(dá)陰性者，伴肉眼可見腹膜轉(zhuǎn)移灶(P1～3)者明顯高于無肉眼可見腹膜轉(zhuǎn)移灶（P0）者，漿膜受侵者明顯高于漿膜未受侵者（P均＜0.05）。結(jié)論兩種方法均適用于胃癌腹腔脫落癌細(xì)胞的診斷或腹膜轉(zhuǎn)移的預(yù)測，端粒酶活性檢測微量癌細(xì)胞的靈敏度優(yōu)于PLC法檢測； 胃癌端粒酶活性與惡性增殖活性密切相關(guān)； 胃癌高增殖活性是漿膜受侵及腹膜轉(zhuǎn)移的重要原因。
目的 探討wt-P53蛋白對人瘢痕疙瘩成纖維細(xì)胞（keloid fibroblasts,KFBs）端粒酶活性的影響；明確在人KFBs中wt-P53蛋白與端粒酶活性之間的相互關(guān)系。方法 將來源于人瘢痕疙瘩組織的KFBs隨機分成兩組，轉(zhuǎn)染組采用腺病毒介導(dǎo)法將野生型wt-p53基因轉(zhuǎn)染至人KFBs；非轉(zhuǎn)染組KFBs未進(jìn)行野生型wt-p53基因轉(zhuǎn)染。轉(zhuǎn)染48 h后，采用間接免疫熒光法和Western blotting法檢測KFBs wt-P53蛋白的表達(dá)；并于轉(zhuǎn)染后1～7 d，采用TRAP-ELISA法檢測KFBs端粒酶活性。結(jié)果 兩組均有wt-P53蛋白表達(dá)，轉(zhuǎn)染組wt-P53蛋白表達(dá)明顯高于非轉(zhuǎn)染組；轉(zhuǎn)染后1～7 d, 轉(zhuǎn)染組端粒酶活性均明顯低于非轉(zhuǎn)染組（P＜0.05結(jié)論 wt-P53蛋白能夠抑制人KFBs端粒酶活性。
【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，并探討其可能機制?！》椒ā〔煌瑵舛華GE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）。 結(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)。【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.