摘要
目的:探讨miR-342与乳腺癌临床病理的关系及是否参与调节ERα并影响他莫昔芬(TAM)的敏感性。方法:RT-PCR检测48例癌组织miR-342和ERαmRNA水平及其中24例癌旁组织miR-342的表达;免疫组化评价癌组织ER、PR、HER-2和VEGF的状态;RT-PCR检测乳腺癌细胞株MCF-7、SKBR-3及MB-231的miR-342、ERαmRNA水平;检测MCF-7细胞瞬时转染hsa-miR342(分mimic、inhibitor及各自阴性对照共4组)48h后miR-342和ERαmRNA的变化;1×10-8 mol/L 17-β雌二醇(E2)单独或联合2×10-5 mol/L TAM处理MCF-7细胞72h,CCK-8法测不同转染组细胞增殖;各转染组细胞1.5×10-5 mol/L TAM处理48h,流式细胞术分析细胞凋亡率。结果:miR-342及ERαmR-NA在ERα阳性乳腺癌组织及细胞中均明显升高,P<0.01;miR-342和ERαmRNA之间存在正相关,P=0.003。miR-342在HER-2阴性(P=0.001)及VEGF阴性(P=0.031)乳腺癌组织中上调;miR-342与PR、淋巴转移、病理分级等因素无明显关系,P>0.05;癌与癌旁miR-342表达差异无统计学意义,P=0.065。MCF-7细胞mimic组ERαmRNA表达为1.80±0.14,高于对照组的1.0±0.0,P=0.001;inhibitor组为0.747±0.087,较对照组低,P=0.037。TAM作用72h,mimic组细胞增殖率为(45.9±1.3)%,与对照组的(55.0±1.5)%相比明显抑制,P=0.001;inhibitor组增殖率为(72.9±1.9)%,较对照组升高,P=0.000。流式细胞术分析TAM处理后的细胞凋亡率,结果显示,mimic组凋亡率为(9.54±1.14)%,较对照组的(4.50±0.46)%增加,P=0.002;inhibitor组凋亡率为(3.06±0.42)%,较对照组的(4.95±0.59)%降低,P=0.011。结论:miR-342的表达可以一定程度预测ERα的表达水平,并作为分子标志预测乳腺癌细胞对TAM的敏感性;miR-342有望成为潜在靶点来参与乳腺癌内分泌治疗。
OBJECTIVE: To investigate the potential relationship between miR-342 and clinicopathological characteristics and examine whether miR-342 adjusts estrogen receptor-α expression level and affects tamoxifen sensitivity of breast cancer.METHODS:The level of miR-342 and ERα mRNA from 48 BRCA cases and expression of miR-342 from 24 normal adjacent tissues were quantitated by real-time RT-PCR;ER,PR,HER-2 and VEGF status were evaluated by immunohistochemical analysis;expression of miR-342,ERα mRNA of breast cancer cells MCF-7,SKBR-3 and MB-231was quantitated by real-time RT-PCR;the change of expression of miR-342 and ERα mRNA was quantitated after MCF-7 cells were transfected by hsa-miR-342-3p mimic,inhibitor and negative control for 48 hours;cell growth was measured by utilizing CCK8 reagent kit after MCF-7 cells were treated with either 1×10-8 mol/L 17-β-estradiol alone or its combination with 2×10-5 mol/L 4-hydroxytamoxifen for 72 hours;apoptosis assay was determined by flowcytometry after all cells were treated with 1.5×10-5 mol/L 4-hydroxytamoxifen for 48 hours.RESULTS: The miR-342 and ERα mRNA expression increased significantly in ERα-positive breast cancer(P<0.01).miR-342 expression was positively correlated with ERα mRNA expression in human breast cancer(P=0.003).miR-342 expression upregulated in HER-2 negative(P=0.001) and VEGF negative(P=0.031) breast cancer;miR-342 expression was of no obvious relevance with PR,lymph node metastasi and pathologic grade(P>0.05);no discrepancy of miR-342 expression existed between cancer and cancer adjacent tissues(P=0.065).Introduction to MCF-7 cells with miR-342 mimic increased ERα mRNA expression(1.80±0.14,P=0.001) and was more sensitive to TAM((45.9±1.3)%,P=0.001) compared with cells transected with negative control,which were 1.0±0.0 and(55.0±1.5)% respectively,while introduction with miR-342 inhibitor decreased ERα mRNA expression(0.747±0.087,P=0.037) and TAM resistance((72.9±1.9)%,P=0.000).Treated with 1.5×10-5 mol/L TAM,apoptosis of MCF-7 increased in
出处
《中华肿瘤防治杂志》
CAS
北大核心
2013年第2期81-87,共7页
Chinese Journal of Cancer Prevention and Treatment
基金
国家自然科学基金(30840093)
2011第四期"333工程"第二层次项目(BRA2011214)
