基于miR-101-5p-ESR1通路探讨职业性镍暴露对结直肠癌SW620细胞恶行生物学行为的影响

摘要/Abstract

摘要： 目的基于微小核糖核酸-101-5p（microRNA-101-5p,miR-101-5p）-雌激素受体1（estrogen receptor,ESR1）通路,探讨职业性镍暴露对结直肠癌SW620细胞恶行生物学行为的影响。方法设立结直肠癌SW620细胞组、镍暴露低、中、高剂量组（NiSO₄最终浓度分别为200.0、400.0、800.0 mg/mL）,于37 °C、5%CO₂、相对湿度（relative humidity,RH）95%的环境下培养72 h,培养结束后噻唑蓝法（methyl thiazolyl tetrazolium,MTT）测定细胞活力、结晶紫染色测定单克隆数目、流式细胞仪检测细胞凋亡、基质胶涂层膜及划痕实验测定细胞侵袭迁移水平、实时荧光定量聚合酶链式反应（polymer ase chain reaction,PCR法及蛋白印记法测定miR-101-5p、ESR1 水平。结果与结直肠癌SW620细胞组比较,NiSO₄暴露低、中、高剂量组存活率水平、克隆形成数目、穿膜数、迁移距离、ESR1 mRNA蛋白表达水平[（1.39±0.22）vs（1.91±0.31）、（3.27±0.50）、（5.89±0.47）,（0.45±0.07）vs（0.88±0.11）、（1.25±0.13）、（1.45±0.18）]升高,凋亡率、miR-101-5p表达水平[（3.58±0.36）vs（2.85±0.21）、（2.29±0.19）、（1.15±0.08）]降低,差异均有统计学意义（均P<0.05）。随着NiSO₄暴露剂量水平的逐渐升高,NiSO₄暴露各剂量组存活率水平、克隆形成数目、穿膜数、迁移距离、ESR1 mRNA蛋白表达水平[（1.91±0.31）vs（3.27±0.50）vs（5.89±0.47）,（0.88±0.11）vs（1.25±0.13）vs（1.45±0.18）]逐渐升高,凋亡率、miR-101-5p表达水平[（2.85±0.21vs（2.29±0.19）vs（1.15±0.08）]逐渐降低,差异均有统计学意义（均P<0.05）。结论镍可抑制结直肠癌细胞miR-101-5p水平表达并促进ESR1高表达,进而促进结直肠癌细胞增殖、侵袭迁移,并抑制凋亡。

关键词: miR-101-5p, ESR1, 职业性, 镍, 结直肠癌, 恶行生物学行为

Abstract: Objective Based on microRNA-101-5p（miR-101-5p）-estrogen receptor（ESR1） pathway,to explore the effect of occupational nickel exposure on malignant biological behavior of colorectal cancer SW620 cells. Methods The colorectal cancer SW620 cell group,low-dose,medium-dose,and high-dose groups exposed to nickel（NiSO₄ final concentrations were 200.0,400.0,and 800.0 mg/mL） were set up in an environment of 37 ℃,5% CO₂,and 95% humidity.After culturing for 72 hours,the cell viability was measured by methyl thiazolyl tetrazolium（MTT） method,the number of single clones was measured by crystal violet staining,the cell apoptosis was detected by flow cytometry,the level of cell invasion and migration was measured by matrigel coating membrane and scratch test,and the levels of miR-101-5p and ESR1 were determined by real time fluorescence quantitative PCR and Western blot. Results Compared with the colorectal cancer SW620 cell group,the survival rate,colony formation number,membrane penetration number,migration distance,ESR1 mRNA protein expression level of low,medium and high dose NiSO₄ exposure groups[（1.39±0.22）vs（1.91±0.31）,（3.27±0.50）,（5.89±0.47） and （0.45±0.07） vs（0.88±0.11）,（1.25±0.13）,（1.45±0.18）] increased,while the apoptosis rate,miR-101-5p expression level[（3.58±0.36） vs（2.85±0.21）,（2.29±0.19）,（1.15±0.08）] decreased,and the differences were statistically significant（all P<0.05）.With the gradual increase of NiSO₄ exposure dose level,the survival rate,colony formation number,membrane penetration number,migration distance,ESR1 mRNA protein expression level of each dose group exposed to NiSO₄[（1.91±0.31） vs（3.27±0.50） vs（5.89±0.47）,（0.88±0.11） vs（1.25±0.13） vs（1.45±0.18）] gradually increased,the apoptosis rate,miR-101-5p expression level[（2.85±0.21） vs（2.29±0.19） vs（1.15±0.08）] gradually decreased,and the differences were statistically significant（all P<0.05）. Conclusion Nickel inhibits the expression of miR-101-5p in colorectal cancer cells and promotes the high expression of ESR1,thereby promoting the proliferation,invasion and migration of colorectal cancer cells,and inhibiting apoptosis.

Key words: miR-101-5p, ESR1, Occupational, Nickel, Colorectal cancer, Malignant biological behavior

中图分类号:

R114

汪星, 张彪. 基于miR-101-5p-ESR1通路探讨职业性镍暴露对结直肠癌SW620细胞恶行生物学行为的影响. [J]职业与健康, 2024, 40(14): 1904-1910.

WANG Xing, ZHANG Biao. Effect of occupational nickel exposure on malignant biological behavior of colorectal cancer SW620 cells：Based on the miR-101-5p-ESR1 pathway. [J]OCCUPATION AND HEALTH, 2024, 40(14): 1904-1910.

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