二氯二苯基三氯乙烷诱导人结直肠癌增殖、侵袭与miR-129/CDK-14表达的关系

职业与健康 ›› 2023, Vol. 39 ›› Issue (19): 2626-2632.

二氯二苯基三氯乙烷诱导人结直肠癌增殖、侵袭与miR-129/CDK-14表达的关系

蒲志忠, 吴万

重庆市开州区人民医院胃肠乳腺外科,重庆 405400

收稿日期:2023-02-11 修回日期:2023-03-14 出版日期:2023-10-01 发布日期:2026-03-26
通信作者: 吴万,主管护师,E-mail：605271180@qq.com
作者简介:蒲志忠,男,主任医师,主要从事胃肠乳腺相关恶性肿瘤的治疗工作。
基金资助:
重庆市卫计委项目（2017MSXM194）

Relationship between proliferation and invasion of human colorectal cancer induced by dichlorodiphenyltrichloroethane and expression of miR-129/CDK-14

PU Zhizhong, WU Wan

Department of Gastrointestinal and Breast Surgery,Kaizhou District People's Hospital,Chongqing 405400,China

Received:2023-02-11 Revised:2023-03-14 Online:2023-10-01 Published:2026-03-26
Contact: WU Wan,Nurse in charge,E-mail：605271180@qq.Com

摘要/Abstract

摘要： 目的探讨二氯二苯基三氯乙烷（dichlorodiphenyltrichloroethane,DDT）诱导人结直肠癌增殖、侵袭与微小RNA-129（microRNA-129,miR-129）/细胞周期蛋白依赖性激酶-14（cyclin-dependent kinase-14,CDK-14）表达的关系。方法 5×10⁶个/mL密度的人结直肠癌细胞分别用终浓度为0、100、250和500 nmol/L 的DDT处理72 h,运用3-（4,5-二甲基噻唑-2）-2,5-二苯基四氮唑溴盐[3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide,MTT]法及甲基紫染色测定细胞活力的和细胞细胞集落,流式细胞仪测定细胞凋亡水平,Transwell小室（transwell chamber,Transwell）侵袭室及划痕试验测定细胞侵袭迁移水平,实时荧光逆转录（real-time fluorescent reverse transcription,RT-PCR）法及蛋白印迹法测定细胞miR-129、CDK-14、卷曲螺旋结构域-34（coiled-coil domain-34,CCDC34）、半乳糖激酶1（Galactokinase 1,GALK1）水平。结果 100、250和500 nmol/L DDT组细胞增殖水平[（70.59±7.72、76.63±8.32、84.25±9.04 vs 62.58±6.83）%]、细胞集落[（99.95±9.58、215.63±19.85、398.96±35.26 vs 62.63±6.52）个]、侵袭数目[（76.25±9.58、154.62±19.51、298.96±38.57 vs 49.65±6.65）个]、迁移水平（75.96±14.58、119.69±20.59、165.63±32.68 vs 11.59±2.28）、CDK-14、CCDC34、GALK1 mRNA蛋白水平明显高于对照组（P<0.01）,凋亡率、miR-129水平明显低于对照组（P<0.01）;且500 nmol/L DDT组细胞增殖水平、细胞集落、侵袭数目、迁移水平、CDK-14 mRNA蛋白[（1.60±0.28、2.14±0.33、2.99±0.51 vs 0.92±0.16）,（0.42±0.07、0.85±0.14、1.20±0.20 vs 0.21±0.03）]、CCDC34（0.48±0.08、0.87±0.15、1.18±0.18 vs 0.28±0.07）、GALK1（0.50±0.09、0.84±0.14、1.22±0.19 vs 0.30±0.08）蛋白水平均高于250 nmol/L DDT组,差异均有统计学意义（均P<0.01）,凋亡率（1.70±0.31、1.00±0.16、0.60±0.11 vs 3.00±0.51）、miR-129（1.42±0.24、0.92±0.15、0.64±0.10 vs 1.78±0.28）水平均低于250 nmol/L DDT组,差异均有统计学意义（均P<0.01）;250 nmol/L DDT组细胞增殖水平、细胞集落、侵袭数目、迁移水平、CDK-14、CCDC34、GALK1 mRNA蛋白水平均高于100 nmol/L DDT组,差异均有统计学意义（均P<0.01）,250 nmol/L DDT组细胞凋亡率、miR-129水平均低于100 nmol/L DDT组,差异均有统计学意义（均P<0.01）;DDT浓度与细胞增殖水平、细胞集落、侵袭数目、迁移水平、凋亡率、miR-129水平、CDK-14、CCDC34、GALK1 mRNA蛋白水平有明显剂量-反应关系（P<0.01）。结论 DDT下调miR-129表达靶向上调调控CDK-14以及CCDC34、GALK1的表达,从而促进结直肠癌细胞的增殖和侵袭,并抑制其凋亡。

关键词: 二氯二苯基三氯乙烷, 结直肠, 微小RNA-129, 细胞周期蛋白依赖性激酶-14

Abstract: Objective To investigate the relationship between the proliferation and invasion of human colorectal cancer induced by dichlorodiphenyltrichloroethane（DDT） and the expression of microRNA-129（miR-129）/cyclin-dependent kinase-14（CDK-14）. Methods The human colorectal cancer cells with a density of 5×10⁶/mL were treated with DDT at final concentrations of 0.0,100,250,and 500 nmol/L for 72 hours,respectively. The cell viability and cell colony were determined by the 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide（MTT） method and methyl violet staining. The apoptosis level was measured by flow cytometry. The invasion and migration levels of cells were measured by transwell chamber（Transwell） and scratch assay. The levels of miR-129,CDK-14,coiled-coil domain-34（CCDC34） and Galactokinase 1（GALK1） were measured by real-time fluorescent reverse transcription（RT-PCR） and Western blotting method. Results The cell proliferation level[（70.59±7.72,76.63±8.32,84.25±9.04 vs 62.58±6.83）%],cell colony（99.95±9.58,215.63±19.85,398.96±35.26 vs 62.63±6.52）,invasion number（76.25±9.58, 154.62±19.51,298.96±38.57 vs 49.65±6.65）,migration level（75.96±14.58,119.69±20.59,165.63±32.68 vs 11.59±2.28）,as well as CDK-14,CCDC34,GALK1 mRNA protein levels in 100,250,500 nmol/L DDT groups were significantly higher than those in the control group（P<0.01）,and the apoptosis rate and miR-129 level were significantly lower than those in the control group（P<0.01）. The cell proliferation level,cell colony,invasion number,migration level,CDK-14 mRNA protein[（1.60±0.28,2.14±0.33,2.99±0.51 vs 0.92±0.16）,（0.42±0.07,0.85±0.14,1.20±0.20 vs 0.21±0.03）],CCDC34（0.48±0.08,0.87±0.15,1.18±0.18 vs 0.28±0.07）,GALK1（0.50±0.09,0.84±0.14,1.22±0.19 vs 0.30±0.08） protein levels in 500 nmol/LDDT group were higher than those in 250 nmol/L DDT group,and the differences were statistically significant（all P<0.01）. The apoptosis rate（1.70±0.31,1.00±0.16, 0.60±0.11 vs 3.00±0.51） and miR-129（1.42±0.24,0.92±0.15,0.64±0.10 vs 1.78±0.28） level were lower than those in 250 nmol/L DDT group,and the differences were statistically significant（all P<0.01）.The cell proliferation level,cell colony,invasion number, migration level,CDK-14,CCDC34,and GALK1 mRNA protein levels in 250 nmol/L DDT group were higher than those in 100 nmol/L DDT group,and the differences were statistically significant（all P<0.01）. The apoptosis rateand miR-129 level in 250 nmol/L DDT group were lower than those in 100 nmol/L DDT group,and the differences were statistically significant（all P<0.01）. There was a significant dose-response relationship between DDT concentration and cell proliferation level,cell colony,invasion number,migration level,apoptosis rate,miR-129 level,CDK-14,CCDC34,GALK1 mRNA protein level（P<0.01）. Conclusion DDT down-regulates the expression of miR-129 and targets to up-regulate the expression of CDK-14,CCDC34 and GALK1,thereby promoting the proliferation and invasion of colorectal cancer cells and inhibiting their apoptosis.

Key words: Dichlorodiphenyltrichloroethane, Colorectum, MicroRNA-129, Cyclin-dependent kinase-14

中图分类号:

R114

蒲志忠, 吴万. 二氯二苯基三氯乙烷诱导人结直肠癌增殖、侵袭与miR-129/CDK-14表达的关系. [J]职业与健康, 2023, 39(19): 2626-2632.

PU Zhizhong, WU Wan. Relationship between proliferation and invasion of human colorectal cancer induced by dichlorodiphenyltrichloroethane and expression of miR-129/CDK-14. [J]OCCUPATION AND HEALTH, 2023, 39(19): 2626-2632.

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