摘要
目的探讨Osteoglycin(OGN)基因是否通过上调雌激素受体(ER)的表达抑制Luminal型乳腺癌细胞增殖。方法 ①通过Ualcan在线数据库分析OGN在乳腺癌样本中的mRNA表达情况,使用Kaplan-Meier Plotter在线分析OGN高或低表达(以OGN在乳腺癌患者中表达的中位数为界限)对乳腺癌患者预后的影响。②实时定量PCR(qRT-PCR)法检测OGN在Luminal型乳腺癌临床组织样本中的表达。③通过转染过表达OGN的质粒上调Luminal型乳腺癌细胞系MCF-7和T47D细胞中OGN的表达水平后再用qRT-PCR和Western blot法检测细胞中OGN和ER表达变化,同时采用CCK8法和克隆形成实验检测过表达OGN的MCF-7和T47D细胞的增殖和克隆形成能力。④采用si RNA瞬时转染干扰过表达OGN的乳腺癌细胞系后再检测ER(ESR1)的表达,同时采用CCK8法检测下调过表达OGN乳腺癌细胞系ER表达后的增殖能力。结果 ①Ualcan在线数据库中,OGN mRNA在不同类型乳腺癌组织中的表达均低于正常乳腺组织(P<0.001)且在Luminal型乳腺癌组织中表达最高(P<0.001)。Kaplan-Meier Plotter预后分析结果显示,在所有乳腺癌或Luminal型乳腺癌患者中,OGN高表达者的预后情况均优于OGN低表达者(P=0.000 14,P=0.001 80)。②临床病例数据中,OGN mRNA在Luminal型乳腺癌组织中低于其相应的癌旁组织(t=4.774,P=0.000 019)。③转染过表达OGN质粒后MCF-7和T47D细胞中OGN和ER表达均上调(P=0.000 002、P=0.000 001);且细胞增殖均被抑制(P<0.05),细胞克隆数明显减少(P<0.05)。④siRNA瞬时转染干扰过表达OGN的乳腺癌细胞系后,ER mRNA水平降低(P<0.05),细胞增殖能力明显增强(P<0.05)。结论 OGN在Luminal型乳腺癌中发挥抑癌作用,其可能机制在于上调ER的表达。
Objective To investigate whether Osteoglycin(OGN) gene inhibits the proliferation of Luminal breast cancer cells by up-regulating the expression of estrogen receptor(ER). Methods① Ualcan online database was used to analyze the expression of OGN in the breast cancer, and Kaplan-Meier Plotter was used to analyze the effect of OGN on the prognosis of patients with breast cancer. The median OGN mRNA expression level was taken as the cut-off point for high or low OGN expression.②The expression of OGN mRNA in the Luminal breast cancer tissue of clinical case was examined using real time quantitative PCR(qRT-PCR).③Up-regulation of OGN expression in the Luminal breast cancer cell lines MCF-7 and T47 D cells by transfection of overexpressing OGN plasmid, the expressions of OGN and ER were detected by qRT-PCR and Western blot, respectively. CCK8 assay and colony formation assay were applied to detect the cell proliferation and colony formation of Luminal breast cancer cells.④siRNA transfection was used to interfere with the expression of ER(ESR1) of breast cancer cells in the overexpressing OGN of breast cancer cells, then the CCK8 assay was used to detect the proliferation ability of the breast cancer cell lines after down-regulating the expression of ER in the overexpressing OGN patients. Results①The results of Ualcan online database showed that the expression of OGN mRNA in the breast cancer tissues of different types of breast cancer was lower than that in the normal breast tissues(P<0.001), and which was highest in the Luminal breast cancer tissues(P<0.001). The Kaplan-Meier Plotter prognosis analysis showed that in all breast cancer or Luminal breast cancer patients, the prognosis of patients with high OGN expression was better than those with low OGN expression(P=0.000 14, P=0.001 80).②The OGN mRNA expression was decreased in the 30 Luminal breast cancer samples as compared with the corresponding adjacent normal breast tissues(t=4.774, P=0.000 019).③The expressions of OGN and ER in the MCF-7 and T47 D cells were up-regulated after transfection of overexpressing OGN plasmid(P=0.000 002, P=0.000 001). The cell proliferation was inhibited(P<0.05) and the number of cell clones was decreased significantly(P<0.05).④After transient transfection of siRNA interfered with breast cancer cell lines of overexpressing OGN, ER mRNA level decreased(P<0.05), and cell proliferation ability increased significantly(P<0.05). Conclusion OGN could exert a tumor suppressor effect in Luminal breast cancer by mediating expression of ER.
作者
章锐
杜秋丽
阮剑
赵建国
ZHANG Rui;DU Qiuli;RUAN Jian;ZHAO Jianguo(Department of Thyroid and Breast Surgery,Wuhan No.1 Hospital,Wuhan 430030,P.R.China)
出处
《中国普外基础与临床杂志》
CAS
2021年第12期1580-1586,共7页
Chinese Journal of Bases and Clinics In General Surgery
基金
湖北省卫生健康科研基金资助项目(项目编号:WJ2021M001)。
