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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 relationship between miR-342 and the clinicopathological features of breast cancer and the role of miR-342 in the regulation of ERα and its impact on the sensitivity of tamoxifen (TAM). Methods: The expression of miR-342 and ERα mRNA in 48 cases of cancerous tissues and the expression of miR-342 in 24 cases of paracancerous tissues were detected by RT-PCR. The status of ER, PR, HER-2 and VEGF in cancer tissues was evaluated by immunohistochemistry. The mRNA levels of miR-342 and ERα in breast cancer cell lines MCF-7, SKBR-3 and MB-231 were detected. The expression of hsa-miR342 in MCF-7 cells transfected with mimic, inhibitor and their negative controls The changes of miR-342 and ERα mRNA in MCF-7 cells treated with 1 × 10-8 mol / L 17-β-estradiol (E2) alone or in combination with 2 × 10-5 mol / L TAM for 72h were determined by CCK-8 Transfection group cells proliferation; transfected cells 1.5 × 10-5 mol / L TAM treatment 48h, flow cytometry analysis of apoptosis rate. Results: The levels of miR-342 and ERαmR-NA in ERα positive breast cancer tissues and cells were significantly increased (P <0.01). There was a positive correlation between miR-342 and ERα mRNA (P = 0.003). miR-342 was up-regulated in HER-2 negative (P = 0.001) and VEGF negative (P = 0.031) breast cancer tissues. There was no significant correlation between miR-342 and PR, lymph node metastasis, pathological grade and other factors There was no significant difference in the expression of miR-342 adjacent to cancer, P = 0.065. The expression of ERαmRNA in MCF-7 cells was 1.80 ± 0.14, which was higher than that of the control group (1.0 ± 0.0, P = 0.001). The inhibitor group was 0.747 ± 0.087, which was lower than that of the control group (P = 0.037). The proliferation rate of mimic group was (45.9 ± 1.3)% at 72h after TAM treatment, which was significantly lower than that of control group (55.0 ± 1.5)%, P = 0.001. The proliferation rate of mimic group was (72.9 ± 1.9)%, Group increased, P = 0.000. Flow cytometry analysis of apoptotic rate after TAM treatment showed that apoptosis rate in mimic group was (9.54 ± 1.14)%, which was significantly higher than that in control group (4.50 ± 0.46)%, P = 0.002; apoptosis in inhibitor group The rate was (3.06 ± 0.42)%, lower than that of the control group (4.95 ± 0.59)%, P = 0.011. CONCLUSIONS: The expression of miR-342 can predict the expression of ERα to a certain extent, and as a molecular marker to predict the sensitivity of breast cancer cells to TAM. MiR-342 is expected to be a potential target to participate in breast cancer endocrine therapy.