| 智会,张志永.利拉鲁肽对结肠癌细胞增殖、凋亡及转录活化因子6通路的影响[J].安徽医药,2020,24(3):438-442. |
| 利拉鲁肽对结肠癌细胞增殖、凋亡及转录活化因子6通路的影响 |
| The influences of liraglutide on proliferation，apoptosis， and ATF6 pathway of colon cancer cells |
| |
| DOI：10.3969/j.issn.1009?6469.2020.03.004 |
| 中文关键词: 结肠肿瘤 基因， ras CCAAT增强子结合蛋白类 半胱氨酸天冬氨酸蛋白酶 原癌基因蛋白质 c?bcl?2 细胞增殖 细胞凋亡 利拉鲁肽 转录活化因子 6通路 |
| 英文关键词: Colonic neoplasms Genes,ras CCAAT?enhancer?binding proteins Caspases Proto?oncogene proteins c?bcl?2 Cell proliferation Apoptosis Liraglutide ATF6 pathway |
| 基金项目: |
|
| 摘要点击次数: 1911 |
| 全文下载次数: 596 |
| 中文摘要: |
| 目的探究利拉鲁肽对结肠癌细胞增殖、凋亡的影响及对转录活化因子 6(ATF6)通路的影响。方法按照利拉鲁肽浓度 0、10、100、1 000 nM培养人结肠癌 HCT116细胞 24、48、72 h，人胆囊收缩素 /缩胆囊素八肽(CCK?8)法检测细胞增殖情况，流式细胞仪检测细胞周期、凋亡情况，蛋白质印迹法(Western Blot)检测作用后细胞中 21KD蛋白(Bax)半胱胺酸蛋白酶 ?3(Caspase?3)及 ATF6通路蛋白 ATF6 p50、CCAAT?增强子结合蛋白( C/EBP)同源蛋白( CHOP)的表达水平。结果、与对照组相比，利拉鲁肽各浓度间增殖率均有显著变化( P＜0.05)同一时间点随着利拉鲁肽浓度的增加， 0、10、100、1 000 nM各组细胞的增殖率分别为( 97.16±35.67)%、(81.69±33.37)%、(76.12，±30.23)%、(70.65±28.89)%；(86.17±33.98)%、(58.68±26.77)%、(39.74±10.67)%、(30.17±9.24)%；(83.82±31.19)%、(38.77±10.19)%、(22.98±6.78)%、(16.63±6.12)%，均呈下降趋势。同一浓度随着时间推移， 24、48、72 h各时间点细胞增殖率分别为( 97.16±35.67)%、(86.17±33.98)%、(83.82±31.19)%；(81.69±33.37)%、(58.68± 26.77)%、(38.77±10.19)%；(76.12±30.23)%、(39.74±10.67)%、(22.98±6.78)%；(70.65±28.89)%、(30.17±9.24)%、(16.63± 6.12)%。不同浓度利拉鲁肽作用于 HCT116细胞 72 h后， 10、100、1 000 nM浓度的利拉鲁肽组的 G2/M期细胞比例较 0 nM组显著减少( P＜0.05)，且随着浓度的增加 G2/M期细胞呈现下降趋势。 0、10、100、1 000 nM利拉鲁肽作用下细胞凋亡率分别为(9.06±0.98)%、(10.45±1.12)%、(13.89±1.54)%、(56.08±14.33)%，依次升高( P＜0.05)。与 0 nM组比较， 10、100、1 000 nM浓度的利拉鲁肽组细胞的凋亡率依次显著升高( P＜0.05)；与 10 nM组比较， 100、1 000 nM组凋亡率依次显著升高( P＜0.05)；与 100 nM组比较， 1 000 nM组凋亡率显著升高( P＜0.05)。蛋白质印迹法结果显示， 10、100、1 000 nM利拉鲁肽作用后细胞中 Bax蛋白表达量分别为( 0.69±0.13)、(0.93±0.24)、(1.19±0.25)显著升高( P＜0.05)1 000 nM组 Bax蛋白表达量显著高于 10 nM组( P＜0.05)，100、1 000 nM两组间差异无统计学意义( P＞005)； Caspase?3蛋白表达量分别为( 0.26±0.06)、(0.76±0.15)、(1.15±0.16)依次显著升高(P＜0.05)，100、1000 nM组Caspase?3蛋白表达量显著高于 10 nM组(P＜0.05)，1 000 nM组Caspase?3蛋白表达量显著高于 100 nM组( P＜0.05)； ATF6 p50蛋白表达量分别为( 0.69±0.11)、(0.96±0.09)、(1.20±0.08)； CHOP蛋白表达量分别为( 0.27±0.06)、(0.79±0.08)、(0.99±0.09)均显著升高( P＜0.05)100、1 000 nM组 ATF6 p50、CHOP蛋白表达量显著高于 10 nM组( P＜0.05)1 000 nM组 ATF6 p50、 CHOP 蛋白表达量显著高于100 nM组( P＜0.05)。结论 利拉鲁肽对结肠癌 HCT116 细胞具有增殖抑制和凋亡促进作用，且呈时间?剂量的依赖性，其可能通过激活 ATF6通路发挥促凋亡作用。 |
| 英文摘要: |
| Objective To investigate the influences of liraglutide on proliferation，apoptosis and ATF6 pathway of colon cancer cells.Methods Human colon cancer HCT116 cells were cultured at liraglutide concentrations of 0，10，100，and 1 000 nM for 24， 48，and 72 h.Cell proliferation was measured by the human cholecystokinin/cholecystokinin octapeptide(CCK?8)method.Cell cycle and apoptosis were detected by cell cytometry，21KD protein(Bax)，caspase?3 and activated transcription factor 6(ATF6)pathwaywere detected by Western Blot Protein ATF6 p50，CCAAT?enhancer binding protein(C/EBP)homologous protein(CHOP)expres? sion levels.Results Compared with the control group，the proliferation rate of all concentrations of liraglutide significantly changed(P＜0.05).At the same time，with the increase of liraglutide concentration，the cell proliferation rate of 0，10，100，and1 000nM group was(97.16±35.67)%，(81.69±33.37)%，(76.12±30.23)%，(70.65±28.89)%；(86.17±33.98)%，(58.68±26.77)%，(39.74± 10.67)%，(30.17±9.24)%；(83.82±31.19)%，(38.77±10.19)%，(22.98±6.78)%，and(16.63±6.12)%，all of which exhibit a down? ward trend.With the same concentration over time，the cell proliferation rate at 24，48，and 72 h point was(97.16±35.67)%，(86.17±33.98)%，(83.82±31.19)%；(81.69±33.37)%，(58.68±26.77)%，(38.77±10.19)%；(76.12±30.23)%，(39.74±10.67)%，(22.98±6.78)%；(70.65±28.89)%，(30.17±9.24)%，(16.63±6.12)%.After 72 h of treatment with different concentrations of liraglu? tide on HCT116 cells，the proportion of G2/M phase cells in 10，100，and 1 000 nM liraglutide groups was significantly lower than that in 0 nM group(P＜0.05)，and G2/M cells showed a decreasing trend with the increase of drug concentration.The apoptosisrate increased under the action of 0，10，100，and 1，000 nM liraglutide were(9.06±0.98)%，(10.45±1.12)%，(13.89±1.54)%，(56.08±14.33)%，respectively(P＜0.05).Compared with the 0 nM group，the apoptosis rate of cells in the liraglutide group at the concentrations of 10，100，and 1 000 nM significantly increased sequentially(P＜0.05).Compared with the 10 nM group，the apop? tosis rate increased significantly(P＜0.05)in the 100，1 000 nM group.Compared with the 100 nM group，the apoptosis rate in the 1，000 nM group increased significantly(P＜0.05).Western blot results showed that Bax protein expression in cells treated with li? raglutide at 10，100，and 1 000 nM was(0.69±0.13)，(0.93±0.24)and(1.19±0.25)，which increased significantly(P＜0.05).Bax protein expression in the 1 000 nM group was significantly higher than that in the 10 nM group(P＜0.05)and there was no signif? icant difference between the two groups(P＞0.05).The expression of Caspase?3 protein was(0.26±0.06).76±0.15)and(1.15±(0，0.16)significantly increased(P＜0.05)，and the expression of Caspase?3 protein in the 100 and 1 000 nM groups was significantly higher than that in the 10 nM group(P＜0.05).The expression of Caspase?3 protein in the 1 000 nM group was significantly high? er than that in the 100 nM group(P＜0.05).The expression levels of ATF6 p50 protein were(0.69±0.11)，(0.96±0.09)and(1.20± 0.08).The expression of CHOP protein was(0.27±0.06)，(0.79±0.08)，and(0.99±0.09)，all significantly increased(P＜0.05).The expression levels of ATF6 p50 and CHOP protein in the 100 and 1，000 nM groups were significantly higher than those in the 10 nM group(P＜0.05).The expression of ATF6 p50 and CHOP protein in the 1 000 nM group was significantly higher than that in the 100 nM group(P＜0.05).Conclusion Liraglutide can inhibit proliferation and promote apoptosis of colon cancer HCT116 cells，and in a time?dose dependent manner，which may play a role in promoting apoptosis by activating ATF6 pathway. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
