目的研究胃癌增殖細胞核抗原(proliferating cell nuclear antigen，PCNA)表達與腹腔灌洗液端粒酶活性及腹膜轉移的相關性，并比較腹腔灌洗液中端粒酶活性和細胞學檢測游離癌細胞預測腹膜轉移的應用價值。方法應用免疫組化SP法檢測60例胃癌患者胃癌組織中PCNA表達,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胞學(peritoneal lavage cytology,PLC)檢測； 并分析其與相關臨床病理因素的關系。結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、組織學類型、浸潤深度、漿膜受累面積及腹膜轉移密切相關，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計學意義，但在未分化型癌、pT4、伴肉眼可見腹膜轉移灶(P1～3)者端粒酶活性陽性率明顯高于PLC。PCNA增殖指數（PI）在腹腔灌洗液端粒酶活性表達陽性者明顯高于表達陰性者，伴肉眼可見腹膜轉移灶(P1～3)者明顯高于無肉眼可見腹膜轉移灶（P0）者，漿膜受侵者明顯高于漿膜未受侵者（P均＜0.05）。結論兩種方法均適用于胃癌腹腔脫落癌細胞的診斷或腹膜轉移的預測，端粒酶活性檢測微量癌細胞的靈敏度優于PLC法檢測； 胃癌端粒酶活性與惡性增殖活性密切相關； 胃癌高增殖活性是漿膜受侵及腹膜轉移的重要原因。
目的 探討wt-P53蛋白對人瘢痕疙瘩成纖維細胞（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系。方法 將來源于人瘢痕疙瘩組織的KFBs隨機分成兩組，轉染組采用腺病毒介導法將野生型wt-p53基因轉染至人KFBs；非轉染組KFBs未進行野生型wt-p53基因轉染。轉染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；并于轉染后1～7 d，采用TRAP-ELISA法檢測KFBs端粒酶活性。結果 兩組均有wt-P53蛋白表達，轉染組wt-P53蛋白表達明顯高于非轉染組；轉染后1～7 d, 轉染組端粒酶活性均明顯低于非轉染組（P＜0.05結論 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.