CAJ | 학술논문

背景：溃疡性结肠炎相关结直肠癌（UcCRC）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 UcCRC 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 UcCRC 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 AuroraB、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）。AuroraB、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 AuroraB、BubR1 MOD 值呈负相关（P ﹤0.05）。AuroraB、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）。AuroraB 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 AuroraB、BubR1表达，继而影响细胞周期有关。
배경：궤양성결장염상관결직장암（UcCRC）시궤양성결장염（UC）최엄중적병발증，5-안기수양산（5-ASA）능강저 UcCRC 적발생풍험。목적：탐토5-ASA 대인결장암세포주 HT-29증식、조망이급세포주기적영향，명학5-ASA대 UcCRC 적억제작용。방법：이불동농도（0、10、20、30、40 mmol/ L）5-ASA 작용우 HT-29세포，채용 CCK-8법검측세포증식정황；채용 TUNEL 법검측세포조망정황；채용류식세포술검측세포주기；채용면역조화법검측세포유사분렬조절인자 AuroraB、BubR1단백표체。결과：5-ASA 10、20、30、40 mmol/ L 조 HT-29세포증식억제솔、조망솔수5-ASA 농도승고이승고（P ﹤0.05）。5-ASA 30、40 mmol/ L 조 G0/ G1기세포비례현저저우0、10、20 mmol/ L 조（P ﹤0.05）；5-ASA 0、10、20、30 mmol/ L 조 S 기세포비례균현저저우40 mmol/ L 조（P ﹤0.05）；5-ASA 30、40 mmol/ L 조G2/ M 기세포비례현저고우0、10、20 mmol/ L 조（P ﹤0.05）。AuroraB、BubR1평균광밀도（MOD）치수5-ASA 농도승고이강저（P ﹤0.05）。5-ASA 농도여세포증식억제솔、조망솔정정상관（P ﹤0.05），여 AuroraB、BubR1 MOD 치정부상관（P ﹤0.05）。AuroraB、BubR1 MOD 치여세포증식억제솔、조망솔、G2/ M 기세포비례정부상관（P ﹤0.05），여G0/ G1기세포비례정정상관（P ﹤0.05）。AuroraB 여 BubR1 MOD 치정정상관（P ﹤0.05）。결론：5-ASA 가억제 HT-29세포증식병유도조망，기궤제가능여5-ASA 억제 AuroraB、BubR1표체，계이영향세포주기유관。
Background:Ulcerative colitis-associated colorectal cancer(UcCRC)is the most serious complication of ulcerative colitis(UC). 5-Aminosalicylic acid(5-ASA)could reduce the risk of UC progressing to UcCRC. 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 UcCRC. 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 AuroraB and BubR1 were determined by immunohistochemistry. Results:Proliferation inhibition rates and apoptosis rates of HT-29 cells in 5-ASA 10,20,30,40 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 AuroraB 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 AuroraB and BubR1( P ﹤ 0. 05). MOD values of AuroraB 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 AuroraB 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 AuroraB and BubR1 and the resulted effect on cell cycle.