中国肿瘤临床
中國腫瘤臨床
중국종류림상
Chinese Journal of Clinical Oncology
2015年
20期
1012-1017
,共6页
乳腺癌%miR-124%SP1%预后%乳腺癌细胞系
乳腺癌%miR-124%SP1%預後%乳腺癌細胞繫
유선암%miR-124%SP1%예후%유선암세포계
breast cancer%miR-124%SP1%prognosis%breast cancer cell line
目的：探讨miR-124表达与乳腺癌发生、发展的相关性及机制。方法：运用实时定量聚合酶链反应（qRT-PCR）检测乳腺癌细胞系以及52例患者乳腺癌癌组织和对应的癌旁正常组织样本中miR-124的表达水平。在乳腺癌细胞株MDA-MB-231和T-47D中过表达miR-124后，测定细胞增殖活性以及侵袭转移能力。构建荧光素酶报告载体pMIR-特异性蛋白1（specificity protein 1，SP1）的3'UTR，利用荧光素酶活性检测鉴定miR-124的预测靶基因SP1。qRT-PCR和Western blot法分别检测SP1的mRNA和蛋白质的表达水平。结果：miR-124在乳腺癌细胞系和癌组织中表达量下调，差异具有统计学意义（P<0.01），并与肿瘤的转移、分期、分级和预后相关。在乳腺癌细胞株MDA-MB-231和T-47D中过表达miR-124后抑制乳腺癌细胞系的增殖、侵袭以及迁移（P<0.01）。转染miR-124模拟物显著抑制荧光素酶的活性（P<0.05）。转染miR-124模拟物显著下调MDA-MB-231和T-47D细胞中SP1的mRNA（P<0.05）和蛋白质的表达水平。结论：miR-124在乳腺癌癌组织中低表达，miR-124低表达与乳腺癌不良预后有关，且miR-124可通过调控转录因子SP1抑制乳腺癌癌细胞的增殖、侵袭和转移。miR-124表达异常减少可能是乳腺癌发生、发展的重要因素。
目的：探討miR-124錶達與乳腺癌髮生、髮展的相關性及機製。方法：運用實時定量聚閤酶鏈反應（qRT-PCR）檢測乳腺癌細胞繫以及52例患者乳腺癌癌組織和對應的癌徬正常組織樣本中miR-124的錶達水平。在乳腺癌細胞株MDA-MB-231和T-47D中過錶達miR-124後，測定細胞增殖活性以及侵襲轉移能力。構建熒光素酶報告載體pMIR-特異性蛋白1（specificity protein 1，SP1）的3'UTR，利用熒光素酶活性檢測鑒定miR-124的預測靶基因SP1。qRT-PCR和Western blot法分彆檢測SP1的mRNA和蛋白質的錶達水平。結果：miR-124在乳腺癌細胞繫和癌組織中錶達量下調，差異具有統計學意義（P<0.01），併與腫瘤的轉移、分期、分級和預後相關。在乳腺癌細胞株MDA-MB-231和T-47D中過錶達miR-124後抑製乳腺癌細胞繫的增殖、侵襲以及遷移（P<0.01）。轉染miR-124模擬物顯著抑製熒光素酶的活性（P<0.05）。轉染miR-124模擬物顯著下調MDA-MB-231和T-47D細胞中SP1的mRNA（P<0.05）和蛋白質的錶達水平。結論：miR-124在乳腺癌癌組織中低錶達，miR-124低錶達與乳腺癌不良預後有關，且miR-124可通過調控轉錄因子SP1抑製乳腺癌癌細胞的增殖、侵襲和轉移。miR-124錶達異常減少可能是乳腺癌髮生、髮展的重要因素。
목적：탐토miR-124표체여유선암발생、발전적상관성급궤제。방법：운용실시정량취합매련반응（qRT-PCR）검측유선암세포계이급52례환자유선암암조직화대응적암방정상조직양본중miR-124적표체수평。재유선암세포주MDA-MB-231화T-47D중과표체miR-124후，측정세포증식활성이급침습전이능력。구건형광소매보고재체pMIR-특이성단백1（specificity protein 1，SP1）적3'UTR，이용형광소매활성검측감정miR-124적예측파기인SP1。qRT-PCR화Western blot법분별검측SP1적mRNA화단백질적표체수평。결과：miR-124재유선암세포계화암조직중표체량하조，차이구유통계학의의（P<0.01），병여종류적전이、분기、분급화예후상관。재유선암세포주MDA-MB-231화T-47D중과표체miR-124후억제유선암세포계적증식、침습이급천이（P<0.01）。전염miR-124모의물현저억제형광소매적활성（P<0.05）。전염miR-124모의물현저하조MDA-MB-231화T-47D세포중SP1적mRNA（P<0.05）화단백질적표체수평。결론：miR-124재유선암암조직중저표체，miR-124저표체여유선암불량예후유관，차miR-124가통과조공전록인자SP1억제유선암암세포적증식、침습화전이。miR-124표체이상감소가능시유선암발생、발전적중요인소。
Objective:To evaluate the role of miR-124 in breast cancer and its underlying mechanism. Methods:Quantitative re-verse transcription-polymerase chain reaction (qRT-PCR) was employed to quantify the expression level of miR-124 in the breast can-cer cell lines and matched tissues of 52 patients. Cell proliferation, invasion, and migration of MDA-MB-231 and T-47D were deter-mined by miR-124 overexpression in vitro. Luciferase vectors (pMIR-SP1 3'UTR) were also constructed. The predicted target gene of miR-124 was identified via luciferase activation assay. The mRNA and protein expression of SP1 was detected via qRT-PCR and West-ern blot, respectively. Results:MiR-124 was decreased in breast cancer tissues and cell lines. This result is correlated with metastatic capacity, TMN stages, and prognosis in breast cancer tissues. In breast cancer cell lines, ectopic overexpression of miR-124 inhibited cell proliferation, invasion, and migration in vitro. MiR-124 mimics significantly inhibited luciferase activation (P<0.05) in HEK293 cells and could significantly decrease the mRNA (P<0.05) and protein expression levels of SP1 in MDA-MB-231 and T-47D cells. Con-clusion:MiR-124 could be inhibited in breast cancer. The low miR-124 expression is associated with poor prognosis. In addition, miR-124 could inhibit cell proliferation, invasion, and migration by targeting SP1. These findings confirm that miR-124 downregulation may be a key mechanism for breast cancer carcinogenesis.
