周佽想，王辰龍，于安路，陳國(guó)強(qiáng)，趙　倩(上海交通大學(xué)醫(yī)學(xué)院，上海200240)

miR-630通過(guò)靶向MTDH抑制乳腺癌進(jìn)程

周佽想，王辰龍，于安路，陳國(guó)強(qiáng)，趙倩△
(上海交通大學(xué)醫(yī)學(xué)院，上海200240)

目的:研究miRNA-630(miR-630)在乳腺癌中的表達(dá)水平及在乳腺癌發(fā)生發(fā)展中的功能。方法:采用反轉(zhuǎn)錄實(shí)時(shí)定量PCR方法對(duì)其在乳腺癌病人標(biāo)本及乳腺癌細(xì)胞系中的表達(dá)進(jìn)行檢測(cè);利用克隆形成實(shí)驗(yàn)、劃痕實(shí)驗(yàn)、細(xì)胞遷移能力檢測(cè)等方法對(duì)其在乳腺癌高轉(zhuǎn)移細(xì)胞株MDA-MB-231及BT549中的功能進(jìn)行研究;利用慢病毒感染方法構(gòu)建穩(wěn)定表達(dá)miR-630的MDA-MB-231細(xì)胞株，研究其在小鼠體內(nèi)轉(zhuǎn)移能力;通過(guò)生物信息學(xué)分析、螢光素酶報(bào)告基因分析和Western blot方法對(duì)miR-630的靶基因進(jìn)行篩選和鑒定。結(jié)果: miR-630在乳腺癌病人癌組織以及乳腺癌細(xì)胞株中的表達(dá)顯著降低;在MDA-MB-231 和BT549細(xì)胞中過(guò)表達(dá)miR-630顯著抑制細(xì)胞的克隆形成能力及遷移侵襲能力。在體內(nèi)實(shí)驗(yàn)中，穩(wěn)定過(guò)表達(dá)miR-630抑制MDA-MB-231細(xì)胞的肺轉(zhuǎn)移能力。最后，MTDH被鑒定為miR-630的靶基因，參與miR-630賦予細(xì)胞的各種功能。結(jié)論: miR-630通過(guò)靶向MTDH在乳腺癌的轉(zhuǎn)移過(guò)程中發(fā)揮重要功能。

Repeated exposure to abuse drugs enhanced the cAMP response element-binding protein (CREB) -regulated cocaine-and amphetamine-regulated transcript (CART) expression in the nucleus accumbens (NAcc)，and these effects was known to contribute to the expression of behavioral sensitization.The present experiments investigated the contribution of microinjection of CART 55-102 into NAcc to the phosphorylation of CREB in this site which is known to participate in the Ca2 +influx and the cAMP-PKA signaling that are regulated responding to caffeine.Here we showed that CART 55-102 inhibited intracellular Ca2 +signal and attenuated caffeine-induced enhancement of depolarization-induced Ca2 +influx in cultured NAcc neurons.Repeated microinjection into the NAcc of CART 55-102 peptide (2.0 g/L per side) blocked the enhancement of p-CREB levels and extracellular signal-regulated kinase (ERK) phosphorylated kinase signaling produced by repeated oral administration of caffeine (30 mg/kg) in this sites at 5 d.Furthermore，the behavioral sensitization was exhibited in the rats given 5-day administration of caffeine following microinjection of saline，but absent in the rats with caffeine following microinjection of CART peptide.These results suggest that phosphorylation of CREB by caffeine in NAcc is blocked by CART 55-102 peptide due to the inhibition of Ca2 +influx and the ERK phosphorylation，and these effects may play a compensatory inhibitory role in expression of behavioral sensitization in the rats receiving microinjection of CART 55-102.

*［Foundation item］Supported by National Natural Science Foundation of China (No.81201035)

△Corresponding author E-mail: huzhenzhen@ncu.edu.cn

Blockade of caffeine-induced CREB phosphorylation by microinjection of CART peptide into nucleus accumbens is linked to inhibition of behavioral sensitization*

SUN Xi，ZHOU Xiao-yan，XU Fang-yun，HU Zhen-zhen△
(Pathophysiology，College of Medicine，Nanchang University，Nanchang 330006，China)

△E-mail: qzhao@shsmu.edu.cn