摘要
目的目前关于tRF-31对乳腺癌的影响研究报道较少。文中旨在检测和分析乳腺癌组织中tRF-31的表达水平,探讨其对乳腺癌细胞增殖的影响。方法收集2017年11月至2019年2月南京医科大学附属肿瘤医院乳腺外科乳腺癌患者的32份肿瘤组织标本及对应的癌旁组织。从中选取6份癌组织以及对应的癌旁组织进行高通量测序,通过高通量测序,选择tRF-31(FC=6.5781,P=0.023)作为研究对象。采用RT-PCR法测定癌组织及癌旁组织的tRF-31表达量,运用ROC曲线分析tRF-31的表达对乳腺癌诊断的灵敏度和特异性。将2种人乳腺癌细胞株(MDA-MB-231和MCF-7)分别分为:对照组(转染negative control)和tRF-31组(转染tRF-31抑制试剂tRF-31 inhibitor)。采用CCK-8和克隆形成实验检测转染后乳腺癌细胞增殖的情况。并测定转染后乳腺癌细胞苏氨酸激酶(AKT)和雷帕霉素靶蛋白(mTOR)表达的变化,探索tRF-31与AKT/mTOR信号通路的关系。结果tRF-31在癌组织(0.103±0.207)的表达显著高于癌旁组织(0.028±0.039),差异有统计学意义(P<0.05)。ROC曲线分析显示,tRF-31检测乳腺癌的灵敏性为90.63%,特异性为53.13%。MDA-MB-231细胞中,tRF-31组tRF-31的表达量较对照组明显降低[(0.267±0.012)vs(1±0.040),P<0.01];MCF-7细胞中,tRF-31组tRF-31的表达量较对照组亦明显降低(P<0.01)。MDA-MB-231和MCF-7细胞中,tRF-31组细胞0、24、48、72 h增殖能力较对照组均降低(P<0.05)。MDA-MB-231细胞中,tRF-31组细胞克隆形成率[(43.67±3.29)%]较对照组[(100±3.74)%]明显降低(P<0.01);MCF-7细胞中,tRF-31组细胞克隆形成率[(49±2.94)%]较对照组[(100±4.89)%]明显降低(P<0.01)。MDA-MB-231和MCF-7细胞中,tRF-31组AKT和mTOR的蛋白水平较对照组明显下降。结论tRF-31在乳腺癌组织中显著高表达,并可促进乳腺癌细胞的增殖。这一结果有望为乳腺癌治疗提供新靶点。
Objective At present,there are few reports on the effect of tRF 31 on breast cancer.This study aims to detect and analyze the expression level of tRF 31 in breast cancer tissues and to explore its effect on the proliferation of breast cancer cells.Methods 32 tumor tissue samples and the corresponding para-cancer tissues from breast cancer patients in The Affiliated Cancer Hospital of Nanjing Medical University from November 2017 to February 2019 were collected.Six out of the thirty two samples were selected for high-throughput sequencing and at last tRF 31(FC=6.5781,P=0.023)was selected as the research object.The expression of tRF-31 in cancer and para-cancer tissues was measured by RT-PCR.The sensitivity and specificity of tRF-31 expression in the breast cancer diagnosis were analyzed by ROC curve.Two kinds of human breast cancer cell lines(MDA-MB-231 and MCF-7)were divided into two groups:control group(transfected with negative control)and tRF-31 group(transfected with tRF-31 inhibitor).The proliferation of transfected breast cancer cells was detected by CCK-8 and clonal formation assay.The expression changes of threonine kinase(AKT)and mammalian target of rapamycin(mTOR)in breast cancer cells after transfection were measured to explore the relationship between TRF-31 and AKT/mTOR signaling pathway.Results The expression of tRF-31 in cancer tissues(0.103±0.207)was significantly higher than that in para-cancerous ones(0.028±0.039).ROC curve showed that the sensitivity and specificity of tRF-31 in detecting breast cancer were 90.63%and 53.13%,respectively.In MDA MB 231 cells,the expression of tRF-31 in the tRF-31 group was significantly lower than that in the control group[(0.267±0.012)vs(1±0.040),P<0.01)],and in MCF-7 cells,the expression of tRF-31 in the tRF-31 group was also significantly lower than that in the control group.In MDA-MB-231 and MCF-7 cells,the proliferation ability of the tRF-31 group was lower than that of the control group at 0 h,24 h,48 h and 72 h.In MDA-MB-231 cells,the clonal formation
作者
江盼
严枫
JIANG Pan;YAN Feng(Department of Clinical Laboratory,The Affiliated Cancer Hospital of Nanjing Medical University&Jiangsu Cancer Hospital&Jiangsu Institute of Cancer Research,Nanjing 210009,Jiangsu,China)
出处
《医学研究生学报》
CAS
北大核心
2020年第7期726-731,共6页
Journal of Medical Postgraduates
基金
国家自然科学基金(81871718)
江苏省研究生科研与实践创新计划项目(KYCX18_1501)。
