基于circ-0072309/miR-26a-5p通路探讨职业性镉暴露诱导结直肠癌细胞恶性生物学行为的机制

摘要/Abstract

摘要：

目的基于环状 RNA-0072309（circular RNA-0072309，circ-0072309）/微小RNA-26a-5p（microRNA-26a-5p，miR-26a-5p）通路探讨职业性镉（Cd）暴露诱导结直肠癌细胞恶性生物学行为的机制。方法 2024年2—5月在保定市第一中心医院进行实验，设结直肠癌细胞SW480组、50 nmol/L CdCl₂组、100 nmol/L CdCl₂组、200 nmol/L CdCl₂组；培养72 h。实验结束后，测定细胞增殖、单克隆数目、凋亡水平、侵袭迁移水平、上皮细胞-间充质转化（epithelial-mesenchymal transition，EMT）水平，RT-PCR法及West-blot法检测细胞circ-0072309、miR-26a-5p、磷酸酯酶与张力蛋白同源物（phosphatase and tensin homolog deleted on chromosome 10，PTEN）水平。结果与结直肠癌细胞SW480组比较，50、100、200 nmol/L CdCl₂组波形蛋白（Vimentin）水平、单细胞克隆形成数目、迁移距离、miR-26a-5p mRNA水平（0.97±0.16 vs 1.63±0.27、2.48±0.41、3.35±0.59）、存活率、穿膜数上升，PTEN mRNA蛋白水平（3.24±0.54 vs 2.63±0.43、1.89±0.32、1.01±0.16，1.28±0.21 vs 0.95±0.16、0.60±0.10、0.23±0.05）、上皮钙黏蛋白（epithelial cadherin，E-cadherin）水平、circ-0072309 mRNA水平（3.59±0.61 vs 2.89±0.48、2.12±0.35、1.35±0.23）、凋亡率下降，差异均有统计学意义（均P<0.05）；且随着CdCl₂暴露剂量的增加，50、100、200 nmol/L CdCl₂组Vimentin水平、单细胞克隆形成数目、迁移距离、miR-26a-5p mRNA水平、存活率、穿膜数呈上升趋势，PTEN mRNA蛋白水平、E-cadherin水平、circ-0072309mRNA水平、凋亡率呈下降趋势，差异均有统计学意义（均P<0.05）。结论职业性Cd暴露通过抑制circ-0072309表达，解除其对miR-26a-5p的吸附，导致miR-26a-5p过表达并靶向下调PTEN基因，激活PI3K/AKT通路，从而以剂量依赖性方式（50~200 nmol/L）促进结直肠癌细胞增殖、侵袭迁移及EMT进程（Vimentin↑/E-cadherin↓），抑制凋亡；本研究首次阐明circ-0072309/miR-26a-5p/PTEN轴在镉致癌中的ceRNA调控机制，为职业镉暴露相关结直肠癌的分子干预和circRNA生物标志物开发提供参考依据。

关键词: circ-0072309, miR-26a-5p, 职业性镉暴露, 结直肠癌, 恶性生物学行为

Abstract:

Objective To explore the mechanism of occupational cadmium exposure-induced malignant biological behavior of colorectal cancer cells based on the circular RNA-0072309（circ-0072309）/microRNA-26a-5p（miR-26a-5p） pathway. Methods From February to May 2024，experiments were conducted at Baoding First Central Hospital. Colorectal cancer cells were divided into SW480 group，50 nmol/L CdCl₂ group，100 nmol/L CdCl₂ group，and 200 nmol/L CdCl₂ group. The cells were cultured for 72 hours. After the experiment，cell proliferation，monoclonal number，and apoptosis level，invasion and migration level，epithelial mesenchymal transition（EMT） level were measured. Circ-0072309，miR-26a-5p，and phosphatase and tensin homolog deleted on chromosome 10（PTEN） levels were detected by RT-PCR and West-blot. Results Compared with the colorectal cancer cell SW480 group，Vimentin levels，number of single cell clones，migration distance，miR-26a-5p mRNA levels（0.97±0.16 vs 1.63±0.27，2.48±0.41，3.35±0.59），the survival rate，and number of membrane penetration of the 50，100，and 200 nmol/L CdCl₂ groups increased，while PTEN mRNA protein levels（3.24±0.54 vs 2.63±0.43，1.89±0.32，1.01±0.16；1.28±0.21 vs 0.95±0.16，0.60±0.10，0.23±0.05），epithelial cadherin（E-cadherin），circ-0072309 mRNA（3.59±0.61 vs 2.89±0.48，2.12±0.35，1.35±0.23），and apoptosis rate decreased，and the differences were statistically significant（all P<0.05）. Moreover，with the increase of CdCl₂ exposure dose，Vimentin levels，number of single cell clones，migration distance，miR-26a-5p mRNA level，survival rate，and number of membrane penetration of the 50，100，and 200 nmol/L CdCl₂ groups showed an upward trend，while PTEN mRNA protein levels，E-cadherin level，circ-0072309 mRNA，and apoptosis rate showed a downward trend，and the differences were statistically significant（all P<0.05）. Conclusion Occupational Cd exposure inhibits the expression of circ-0072309 and releases its adsorption to miR-26a-5p，leading to overexpression of miR-26a-5p and targeted down regulation of PTEN gene，activating PI3K/AKT pathway，thereby promoting the proliferation，invasion and migration of colorectal cancer cells and EMT process（Vimentin↑/E-cadherin↓） in a dose-dependent manner（50-200 nmol/L），and inhibiting apoptosis. This study is the first to clarify the ceRNA regulatory mechanism of circ-0072309/miR-26a-5p/PTEN axis in cadmium carcinogenesis，providing a theoretical basis for molecular intervention of occupational cadmium exposure-related colorectal cancer and the development of circRNA biomarkers.

Key words: circ-0072309, miR-26a-5p, Occupational cadmium exposure, Colorectal cancer, Malignant biological behavior

中图分类号:

R735.3

辛清旺, 张培军. 基于circ-0072309/miR-26a-5p通路探讨职业性镉暴露诱导结直肠癌细胞恶性生物学行为的机制. [J]职业与健康, 2025, 41(24): 3350-3356.

XIN Qingwang, ZHANG Peijun. Exploring the mechanism of occupational cadmium exposure inducing malignant biological behavior of colorectal cancer cells based on the circ-0072309/miR-26a-5p pathway. [J]OCCUPATION AND HEALTH, 2025, 41(24): 3350-3356.

图/表 11

参考文献 19

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组别	存活率（%）	单细胞克隆形成数目（个）	凋亡率（%）
200 nmol/L CdCl₂组	93.20±6.34^abc	155.35±15.1^abc	1.90±0.09^abc
100 nmol/L CdCl₂组	84.38±5.89^ab	98.25±10.25^ab	3.60±0.19^ab
50 nmol/L CdCl₂组	73.45±5.38^a	79.29±8.88^a	16.90±0.56^a
结直肠癌细胞SW480组	61.83±4.39	53.83±6.32	18.00±0.78
F值	109.415	165.216	189.554
P	<0.01	<0.01	<0.01

组别	存活率（%）	单细胞克隆形成数目（个）	凋亡率（%）
200 nmol/L CdCl₂组	93.20±6.34^abc	155.35±15.1^abc	1.90±0.09^abc
100 nmol/L CdCl₂组	84.38±5.89^ab	98.25±10.25^ab	3.60±0.19^ab
50 nmol/L CdCl₂组	73.45±5.38^a	79.29±8.88^a	16.90±0.56^a
结直肠癌细胞SW480组	61.83±4.39	53.83±6.32	18.00±0.78
F值	109.415	165.216	189.554
P	<0.01	<0.01	<0.01

组别	穿膜数（个）	迁移距离（μm）
200 nmol/L CdCl₂组	1852.63±302.52^abc	152.66±23.84^abc
100 nmol/L CdCl₂组	1025.58±159.96^ab	89.96±12.59^ab
50 nmol/L CdCl₂组	859.96±142.59^a	43.55±7.36^a
结直肠癌细胞SW480组	259.96±42.59	23.69±3.98
F值	398.654	254.239
P	<0.01	<0.01

组别	穿膜数（个）	迁移距离（μm）
200 nmol/L CdCl₂组	1852.63±302.52^abc	152.66±23.84^abc
100 nmol/L CdCl₂组	1025.58±159.96^ab	89.96±12.59^ab
50 nmol/L CdCl₂组	859.96±142.59^a	43.55±7.36^a
结直肠癌细胞SW480组	259.96±42.59	23.69±3.98
F值	398.654	254.239
P	<0.01	<0.01

组别	E-cadherin蛋白（μmol/L）	Vimentin蛋白（μmol/L）
200 nmol/L CdCl₂组	105.69±15.59^abc	505.25±94.69^abc
100 nmol/L CdCl₂组	217.69±35.22^ab	386.69±65.69^ab
50 nmol/L CdCl₂组	345.14±58.52^a	255.63±42.29^a
结直肠癌细胞SW480组	459.65±75.25	159.96±25.69
F值	156.324	204.329
P	<0.01	<0.01