tRNA衍生物tRF-5026a在乳腺癌患者血清中的水平及其对乳腺癌细胞增殖、侵袭和迁移影响的实验研究
Level of tRNA derivative tRF-5026a in serum of breast cancer patients and experimental study of its effect on proliferation,invasion and migration of breast cancer cells
李聪 1王承霞 1莫冬萍1
作者信息
- 1. 江苏省肿瘤医院 南京医科大学附属肿瘤医院 江苏省肿瘤防治研究所检验科,南京 210009
- 折叠
摘要
目的 探讨转运RNA(tRNA)衍生物tRF-5026a在乳腺癌患者血清中的水平及对乳腺癌诊断的价值,并探讨其体外对乳腺癌细胞生物学功能的影响及可能机制.方法 回顾性选择2016年1月至2019年2月江苏省肿瘤医院住院的60例女性乳腺癌患者(乳腺癌组)及同期20例体检健康女性(健康对照组),采用实时定量聚合酶链反应(RT-qPCR)法检测研究对象血清tRF-5026a的相对表达量;以病理诊断为金标准,绘制依据血清tRF-5026a水平诊断乳腺癌的受试者工作特征(ROC)曲线.采用脂质体法将tRF-5026a模拟物(tRF-5026a组)及阴性对照序列(阴性对照组)瞬时转染至人乳腺癌MCF-7细胞和BT549细胞;采用CCK-8法和5-乙炔基-2-脱氧尿苷(EdU)荧光染色检测各组细胞增殖能力,流式细胞术检测各组细胞凋亡水平,Transwell实验检测各组细胞的侵袭和迁移能力,蛋白质印迹法检测各组细胞中上皮间质转化相关蛋白的表达水平.结果 健康对照组和乳腺癌组血清中tRF-5026a相对表达量·[M(Q,,Q3)]分别为 16.58(6.37,26.31)和3.46(0.32,9.01),差异有统计学意义(Z=-4.27,P＜0.001).ROC曲线分析显示,以血清tRF-5026a相对表达量诊断乳腺癌的曲线下面积(AUC)为0.820(95％CI:0.722～0.918),其最佳临界值为9.082,对应的灵敏度和特异度分别为75.0％和76.7％.tRF-5026a组和阴性对照组MCF-7细胞凋亡率分别为(16.52±0.51)％、(12.28±1.75)％,BT549细胞分别为(13.27±2.18)％、(8.86±0.29)％,差异均无统计学意义(均P＞0.05).tRF-5026a组MCF-7细胞和BT549细胞的增殖能力以及侵袭和迁移能力均低于相对应的阴性对照组细胞(均P＜0.05).tRF-5026a组MCF-7细胞和BT549细胞神经钙黏素及基质金属蛋白酶(MMP)-9和MMP-3蛋白水平均低于相对应的阴性对照组细胞.结论 tRF-5026a在乳腺癌患者血清中水平较低,其可能为乳腺癌诊断的一个指标.体外tRF-5026a能抑制乳腺癌MCF-7细胞和BT549细胞增殖、侵袭和迁移,可能与调控上皮间质转化相关.
Abstract
Objective To investigate the level of the transporter RNA(tRNA)derivative tRF-5026a in the serum of breast cancer patients and its value for the diagnosis of breast cancer,and to investigate its effect on the biological functions of breast cancer cells in vitro and the possible mechanisms.Methods Sixty female breast cancer patients(breast cancer group)hospitalized in Jiangsu Cancer Hospital from January 2016 to February 2019 and 20 healthy women undergoing physical examination during the same period(healthy control group)were retrospectively selected.The relative expression of serum tRF-5026a in the study subjects was detected by real-time quantitative polymerase chain reaction(RT-qPCR).The receiver operating characteristic(ROC)curve of serum tRF-5026a level for the diagnosis of breast cancer was drawn with pathological diagnosis as the gold standard.tRF-5026a mimics(tRF-5026a group)and negative control sequences(negative control group)were transiently transfected into MCF-7 and BT549 cells by lipofectamine method;CCK-8 assay and 5-ethynyl-2-deoxyuridine(EdU)assay were used to detect the ability of cell proliferation in cells of each group;cell apoptosis in cells of each group was detected by flow cytometry;the abilities of cell invasion and migration in cells of each group were detected by Transwell assay;the expressions of epithelial mesenchymal transition-related proteins in cells of each group were detected by Western blotting.Results The relative expressions of tRF-5026a[M(Qv Q3)]in serum of healthy control group and breast cancer group were 16.58(6.37,26.31)and 3.46(0.32,9.01),with a statistically significant difference(Z=-4.27,P＜0.001).ROC curve analysis showed that the area under the curve(AUC)for diagnosis of breast cancer by the relative expression of serum tRF-5026a was 0.820(95％CI:0.722-0.918),with an optimal cut-off value of 9.082,and the corresponding sensitivity and specificity were 75.0％and 76.7％,respectively.The apoptosis rates of MCF-7 cells in the tRF-5026a group and the corresponding negative control group were(16.52±0.51)％and(12.28±1.75)％,and the BT549 cells were(13.27±2.18)％and(8.86±0.29)％,the differences were not statistically significant(both P＞0.05).MCF-7 and BT549 cells in the tRF-5026a group had lower proliferative,invasive and migratory abilities than cells in the corresponding negative control group(all P＜0.05).MCF-7 and BT549 cells in the tRF-5026a group had lower protein expressions of N-cadherin,matrix metalloproteinase(MMP)-9 and MMP-3 than cells in the corresponding negative control group.Conclusions tRF-5026a has low level in the serum of breast cancer patients and it may be an indicator for breast cancer diagnosis.tRF-5026a can inhibit the proliferation,invasion and migration of breast cancer MCF-7 and BT549 cells in vitro,which may be related to the regulation of epithelial mesenchymal transition.
关键词
乳腺肿瘤/转运RNA衍生物/生物学标记/细胞增殖/细胞运动Key words
Breast neoplasms/Transporter RNA derivatives/Biological markers/Cell proliferation/Cell movement引用本文复制引用
出版年
2024
NETL
NSTL
万方数据