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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、100μg/mL小牛血清白蛋白(bovine serum albumin,BSA)组及50、100、500μg/mL AGE组,其余检测只设置100μg/mL BSA组和100μg/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。100μg/mL BSA组与100μg/mL AGE组72h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(P<0.05)。蛋白质印迹法示100μg/mL AGE组72h后CyclinD1的表达较100μg/mL BSA组增加,差异有统计学意义(P<0.05)。TRAP银染法检测示100μg/mL AGE干预SW-480细胞72h后可以增加端粒酶活性(P<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。
Objective To observe the effect of advanced glycosylation end prodrcts (AGE) on the proliferation of human colon carcinoma cell line SW-480 and to explore its possible mechanism. Methods The effect of AGE on the cell cycle of SW-480 cells was observed by MTT assay. The effect of AGE on the cell cycle of SW-480 cells was observed by flow cytometry. The effect of AGE on the expression of CyclinD1 in SW-480 cells was observed by Western blotting Effect of AGE on telomerase activity in SW-480 cells was detected by silver staining with telomeric repeat amplification protocol (TRAP). MTT test cell viability set blank control group, 100μg / mL bovine serum albumin (BSA) group and 50,100,500μg / mL AGE group, the rest of the test set only 100μg / mL BSA group and 100μg / mL AGE group. Results MTT results showed that AGE promoted the proliferation of SW-480 cells in a concentration-dependent manner. The percentages of G0 / G1 phase cells in 100μg / mL BSA group and 100μg / mL AGE group after 72h were 56.02% ± 0.58% and 51.93% ± 1.01%, respectively. The difference was statistically significant (P <0.05). Western blotting showed that the expression of CyclinD1 in 100μg / mL AGE group increased more than 100μg / mL BSA group at 72h (P <0.05). TRAP silver staining showed that telomerase activity could be increased after SWG 480 cells were treated with 100μg / mL AGE for 72h (P <0.05). Conclusion AGE can promote the growth of human colon cancer cell SW-480 in a dose-dependent manner. Its mechanism may be related to AGE up-regulation of CyclinD1 expression and accelerated G1 / S phase transition and increased telomerase activity.