【摘要】 背景:溃疡性结肠炎相关结直肠癌(Uc CRC)是溃疡性结肠炎(UC)最严重的并发症,5-氨基水杨酸(5-ASA)能降低Uc CRC的发生风险。目的:探讨5-ASA对人结肠癌细胞株HT-29增殖、凋亡以及细胞周期的影响,明确5-ASA对Uc CRC的抑制作用。方法:以不同浓度(0、10、20、30、40 mmol/L)5-ASA作用于HT-29细胞,采用CCK-8法检测细胞增殖情况;采用TUNEL法检测细胞凋亡情况;采用流式细胞术检测细胞周期;采用免疫组化法检测细胞有丝分裂调节因子Aurora B、BubR1蛋白表达。结果:5-ASA 10、20、30、40 mmol/L组HT-29细胞增殖抑制率、凋亡率随5-ASA浓度升高而升高(P<0.05)。5-ASA 30、40 mmol/L组G0/G1期细胞比例显著低于0、10、20 mmol/L组(P<0.05);5-ASA 0、10、20、30 mmol/L组S期细胞比例均显著低于40 mmol/L组(P<0.05);5-ASA 30、40 mmol/L组G2/M期细胞比例显著高于0、10、20 mmol/L组(P<0.05)。Aurora B、BubR1平均光密度(MOD)值随5-ASA浓度升高而降低(P<0.05)。5-ASA浓度与细胞增殖抑制率、凋亡率呈正相关(P<0.05),与Aurora B、BubR1 MOD值呈负相关(P<0.05)。Aurora B、BubR1 MOD值与细胞增殖抑制率、凋亡率、G2/M期细胞比例呈负相关(P<0.05),与G0/G1期细胞比例呈正相关(P<0.05)。Aurora B与BubR1 MOD值呈正相关(P<0.05)。结论:5-ASA可抑制HT-29细胞增殖并诱导凋亡,其机制可能与5-ASA抑制Aurora B、BubR1表达,继而影响细胞周期有关。更多还原

【Abstract】 Background: Ulcerative colitis-associated colorectal cancer( Uc CRC) is the most serious complication of ulcerative colitis( UC). 5-Aminosalicylic acid( 5-ASA) could reduce the risk of UC progressing to Uc CRC. Aims: To investigate the effect of 5-ASA on cell proliferation,apoptosis and cell cycle in human colon cancer cell line HT-29 for verifying the inhibitory effect of 5-ASA on Uc CRC. Methods: HT-29 cells were treated with different concentrations( 0,10,20,30,40 mmol / L) of 5-ASA. Cell proliferation was measured by CCK-8 assay. Cell apoptosis was determined by TUNEL method. Cell cycle was analyzed by flow cytometry. Expressions of cell mitotic regulators Aurora B and BubR1 were determined by immunohistochemistry. Results: Proliferation inhibition rates and apoptosis rates of HT-29 cells in 5-ASA10,20,30,40 mmol / L groups were increased with increase in concentration of 5-ASA( P < 0. 05). Proportions of cells in phase G0 / G1 in 5-ASA 30,40 mmol / L groups were significantly lower than those in 5-ASA 0,10,20 mmol / L groups( P < 0. 05); proportions of cells in phase S in 5-ASA 0,10,20,30 mmol/L groups were significantly lower than that in 5-ASA 40 mmol / L group( P < 0. 05); while proportions of cells in phase G2 / M in 5-ASA 30,40 mmol / L groups were significantly higher than those in 5-ASA 0,10,20 mmol / L groups( P < 0. 05). Mean optical density( MOD) values of Aurora B and BubR1 were decreased with increase in concentration of 5-ASA( P < 0. 05). Concentration of 5-ASA was positively correlated with proliferation inhibition rate and apoptosis rate of HT-29 cells( P < 0. 05),but was negatively correlated with MOD values of Aurora B and BubR1( P < 0. 05). MOD values of Aurora B and BubR1 were negatively correlated with proliferation inhibition rate and apoptosis rate of HT-29 cells as well as proportion of cells in phase G2 / M( P <0. 05),but were positively correlated with proportion of cells in phase G0/G1( P <0. 05). MOD value of Aurora B was positively correlated with MOD value of BubR1( P < 0. 05). Conclusions: 5-ASA can inhibit proliferation and induce apoptosis in HT-29 cells,the mechanism may be related to inhibiting expressions of Aurora B and BubR1 and the resulted effect on cell cycle.更多还原