MicroRNA-150-5p對(duì)肝細(xì)胞癌細(xì)胞增殖、凋亡和周期的影響

趙月秋，張正寶，郭紅霞，詹傳飛，魯 皓，錢(qián)曉峰

MicroRNA-150-5p對(duì)肝細(xì)胞癌細(xì)胞增殖、凋亡和周期的影響

趙月秋1，張正寶1，郭紅霞1，詹傳飛2，魯 皓2，錢(qián)曉峰2

（1.江蘇聯(lián)合職業(yè)技術(shù)學(xué)院南京衛(wèi)生分院，江蘇 南京 210038；2.江蘇省人民醫(yī)院 肝臟外科，江蘇 南京 210029）

目的 研究microRNA-150-5p（miR-150-5p）在人正常肝細(xì)胞和肝細(xì)胞癌細(xì)胞中的表達(dá)，及其對(duì)肝細(xì)胞癌細(xì)胞增殖、凋亡和周期的影響及潛在機(jī)制。方法 通過(guò)實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)（qRT-PCR）檢測(cè)各處理組細(xì)胞中的miR-150-5p的表達(dá)水平；MTT法檢測(cè)miR-150-5p對(duì)肝細(xì)胞癌細(xì)胞增殖的影響；流式細(xì)胞術(shù)檢測(cè)miR-150-5p對(duì)肝細(xì)胞癌細(xì)胞凋亡和周期的影響；熒光素酶報(bào)告基因?qū)嶒?yàn)檢測(cè)鋅指蛋白609是否為miR-150-5p的直接靶基因。結(jié)果 miR-150-5p在肝細(xì)胞癌細(xì)胞系中的表達(dá)較人正常肝細(xì)胞細(xì)胞系降低，miR-150-5p可抑制肝細(xì)胞癌細(xì)胞的增殖，促進(jìn)肝細(xì)胞癌細(xì)胞的凋亡，并阻滯細(xì)胞周期于G1期。miR-150-5p在肝細(xì)胞癌中發(fā)揮腫瘤抑制分子作用的潛在機(jī)制為直接靶向抑制ZNF609的表達(dá)。結(jié)論 miR-150-5p在肝細(xì)胞癌細(xì)胞中表達(dá)降低，并可通過(guò)調(diào)控ZNF609的表達(dá)而調(diào)控肝細(xì)胞癌細(xì)胞增殖、凋亡和周期。

MicroRNA-150-5p；增殖；凋亡；周期；ZNF609

在全球范圍內(nèi)，肝細(xì)胞癌發(fā)病率高居所有惡性腫瘤的第5位，值得注意的是，近年來(lái)肝細(xì)胞癌在東亞和發(fā)展中的發(fā)病率正逐步上升[1-2]。盡管在過(guò)去數(shù)10年間，肝細(xì)胞癌的治療手段取得巨大的進(jìn)步，但大多數(shù)患者術(shù)后發(fā)生復(fù)發(fā)或遠(yuǎn)處轉(zhuǎn)移，導(dǎo)致肝細(xì)胞癌的致死率仍居高不下[3]。截止目前，肝細(xì)胞癌發(fā)生、進(jìn)展中的分子機(jī)制仍不完全清楚。因此，尋找與肝細(xì)胞癌發(fā)生及進(jìn)展相關(guān)的分子，對(duì)提高肝細(xì)胞癌患者術(shù)后生存率至關(guān)重要。

MicroRNA（miRNA）是一類可在轉(zhuǎn)錄后水平調(diào)控基因表達(dá)的18-22個(gè)核苷酸大小的非編碼小分子。miRNA在進(jìn)化過(guò)程中發(fā)育相近的物種之間高度保守，被視為細(xì)胞增殖、細(xì)胞分化和腫瘤發(fā)生的關(guān)鍵調(diào)控分子[4-6]。在肝細(xì)胞癌中，多種miRNA被發(fā)現(xiàn)可作為癌基因或抑癌基因參與調(diào)控肝細(xì)胞癌的發(fā)生、進(jìn)展，如miR-214[7]、miR-182[8]、miR-29b[9]、miR-7等[10]。研究發(fā)現(xiàn)，miR-150-5p在肝細(xì)胞癌組織中表達(dá)下調(diào)，并可抑制肝細(xì)胞癌細(xì)胞的侵襲和轉(zhuǎn)移[11]，但miR-150-5p對(duì)肝細(xì)胞癌細(xì)胞增殖、凋亡和周期的影響及潛在分子機(jī)制，目前尚不清楚。本研究擬采用實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)（quantitative real-time polymerasechainreaction，qRT-PCR）方法檢測(cè)miR-150-5p在正常肝細(xì)胞系和肝細(xì)胞癌細(xì)胞系中的表達(dá)差異，及其對(duì)肝細(xì)胞癌細(xì)胞增殖、凋亡和周期的影響。

1 材料與方法

1.1 一般資料

培養(yǎng)細(xì)胞總RNA提取試劑盒購(gòu)自天根生化科技（北京）有限公司，Quant一步法逆轉(zhuǎn)錄-熒光定量試劑盒購(gòu)自天根生化科技（北京）有限公司，miR-150-5p正常對(duì)照（normal control，NC）和miR-150-5p類似物（Mimics）由上海吉瑪公司設(shè)計(jì)并合成，脂質(zhì)體2000（LipofectamineTM2 000）試劑購(gòu)于美國(guó)Invitrogen公司，MTT試劑盒購(gòu)自美國(guó)Promega公司，流式細(xì)胞術(shù)所用FITC標(biāo)記的Annexin-V及PI試劑購(gòu)自美國(guó)BioLegend公司，人正常肝細(xì)胞系QZG細(xì)胞和HL-7702細(xì)胞，肝細(xì)胞癌細(xì)胞系HepG2細(xì)胞和SMMC-7221細(xì)胞均購(gòu)自中國(guó)科學(xué)院上海細(xì)胞庫(kù)。

1.2 方法

1.2.1 細(xì)胞培養(yǎng)和轉(zhuǎn)染 人正常肝細(xì)胞和肝細(xì)胞癌細(xì)胞系用含100 ml/L胎牛血清的1 640培養(yǎng)液，置于溫度37℃，50 ml/L二氧化碳CO2細(xì)胞培養(yǎng)箱中培養(yǎng)。當(dāng)肝細(xì)胞癌細(xì)胞密度達(dá)到約70%后，將miR-150-5p NC和miR-150-5p類似物經(jīng)LipofectamineTM2 000轉(zhuǎn)入細(xì)胞中，具體操作步驟參見(jiàn)脂質(zhì)體2000（LipofectamineTM2 000）試劑說(shuō)明書(shū)。

1.2.2 qRT-PCR 按照培養(yǎng)細(xì)胞總RNA提取試劑盒說(shuō)明書(shū)所述方法提取各實(shí)驗(yàn)組細(xì)胞，而后采用Quant一步法逆轉(zhuǎn)錄-熒光定量試劑盒所述方法行qRT-PCR，具體步驟如下：按說(shuō)明書(shū)所述依次加入試劑后行一步法qRT-PCR。qRT-PCR反應(yīng)條件為：50℃30 min，95℃2 min，35個(gè)循環(huán)94℃20 s，60℃20 s，65℃20 s。

1.2.3 MTT 細(xì)胞轉(zhuǎn)染48 h后，取對(duì)數(shù)生長(zhǎng)期的肝細(xì)胞癌細(xì)胞，制備單細(xì)胞懸液，以5.0×103個(gè)/孔細(xì)胞密度接種于96孔板，每組設(shè)8個(gè)復(fù)孔，分別培養(yǎng)1～5 d，每孔加入20 μl濃度為1.5 g/L的MTT，培養(yǎng)4 h后每孔加入150 μl DMSO，讀取吸光度值，繪制細(xì)胞生長(zhǎng)曲線。

1.2.4 流式細(xì)胞術(shù) 細(xì)胞周期檢測(cè)方法簡(jiǎn)述如下，取轉(zhuǎn)染48h后的各組細(xì)胞，5ml PBS洗滌，1 000 r/min離心5 min，5 ml PBS洗滌3次后，棄上清液；加入100μl流式洗液，PI（50μg/ml）10μl，避光反應(yīng)10 min后，加入400μl流式洗液，200目尼龍膜過(guò)濾細(xì)胞，立即利用流式細(xì)胞儀上機(jī)檢測(cè)。如檢測(cè)細(xì)胞凋亡，加入抗體步驟時(shí)同時(shí)加入FITC標(biāo)記的Annexin-V和PI各10μl及同型對(duì)照抗體，其余步驟同檢測(cè)細(xì)胞周期。細(xì)胞周期和細(xì)胞凋亡數(shù)據(jù)后期均采用Flowjo 7.2軟件進(jìn)行分析。

1.3 統(tǒng)計(jì)學(xué)方法

數(shù)據(jù)分析采用SPSS 17.0統(tǒng)計(jì)軟件，計(jì)量資料以均數(shù)±標(biāo)準(zhǔn)差（x±s）表示，各組間的數(shù)據(jù)比較采用t檢驗(yàn)，P＜0.05為差異有統(tǒng)計(jì)學(xué)意義。

2 結(jié)果

2.1 肝細(xì)胞癌細(xì)胞和正常肝細(xì)胞中miR-150-5p的表達(dá)差異

為研究人正常肝細(xì)胞系與肝癌細(xì)胞系中miR-150-5p的表達(dá)情況，分別提取人正常肝細(xì)胞系QZG和 HL-7702細(xì)胞和肝細(xì)胞癌細(xì)胞系HepG2和SMMC-7221細(xì)胞的總RNA，采用一步法qRT-PCR檢測(cè)各細(xì)胞中miR-150-5p的相對(duì)表達(dá)量，經(jīng)方差分析，4組細(xì)胞中miR-150-5p表達(dá)差異有統(tǒng)計(jì)學(xué)意義（F=9.125，P=0.018），提示miR-150-5p在4組細(xì)胞系中表達(dá)差異有統(tǒng)計(jì)學(xué)意義。進(jìn)一步t檢驗(yàn)分析兩兩細(xì)胞系中miR-150-5p的表達(dá)差異，結(jié)果顯示肝細(xì)胞癌細(xì)胞系中的miR-150-5p的表達(dá)水平較正常肝細(xì)胞降低（QZG vs HL-7702，t=0.324，P=0.419；QZG vs HepG2，t=7.748，P=0.031；QZG vs SMMC-7221，t= 5.197，P=0.039），結(jié)果提示，miR-150-5p在肝細(xì)胞癌中表達(dá)降低，是潛在的腫瘤抑制分子。見(jiàn)圖1。

2.2 miR-150-5p mimics上調(diào)肝細(xì)胞癌細(xì)胞中的miR-150-5p的表達(dá)

為研究miR-150-5p對(duì)肝細(xì)胞癌增殖、凋亡、周期的影響，合成miR-150-5p mimics，并轉(zhuǎn)染至細(xì)胞中觀察miR-150-5p mimics是否可以上調(diào)肝細(xì)胞癌細(xì)胞中的miR-150-5p的表達(dá)水平。轉(zhuǎn)染miR-150-5p NC的肝細(xì)胞癌細(xì)胞命名為miR-150-5p NC組，轉(zhuǎn)染miR-150-5p mimics的肝細(xì)胞癌細(xì)胞命名為miR-150-5p mimics組。轉(zhuǎn)染48 h后，分別提取各組細(xì)胞總RNA，采用一步法qRT-PCR檢測(cè)各組細(xì)胞中miR-150-5p的相對(duì)表達(dá)量，結(jié)果顯示miR-150-5p mimics組的miR-150-5p的相對(duì)表達(dá)量較miR-150-5p NC組上調(diào)（HepG2細(xì)胞：t=5.114，P=0.017；SMMC-7221細(xì)胞：t=3.716，P=0.032），結(jié)果提示miR-150-5p可上調(diào)肝細(xì)胞癌細(xì)胞中miR-150-5p的表達(dá)水平。見(jiàn)圖2。

圖2 miR-150-5p在miR-150-5p NC組和miR-150-5p mimics組肝細(xì)胞癌細(xì)胞中的表達(dá)水平

2.3 miR-150-5p對(duì)肝細(xì)胞癌細(xì)胞增殖的影響

為研究miR-150-5p對(duì)肝細(xì)胞癌細(xì)胞增殖能力的影響，利用MTT實(shí)驗(yàn)檢測(cè)轉(zhuǎn)染miR150-5p-NC與miR-150-5p mimics的肝細(xì)胞癌 HepG2細(xì)胞和SMMC-7221細(xì)胞的細(xì)胞增殖的差異。結(jié)果顯示，從檢測(cè)后第2天開(kāi)始，miR-150-5p mimics組的肝細(xì)胞癌細(xì)胞增殖較miR-150-5p NC組出現(xiàn)明顯的生長(zhǎng)抑制（HepG2細(xì)胞：t=4.146，P=0.022；SMMC-7221細(xì)胞：t=3.193，P=0.041），結(jié)果提示，提高肝細(xì)胞癌細(xì)胞中miR-150-5p的表達(dá)可抑制肝細(xì)胞癌細(xì)胞的增殖。見(jiàn)圖3。

2.4 miR-150-5對(duì)肝細(xì)胞癌細(xì)胞凋亡的影響

為研究miR-150-5p對(duì)肝細(xì)胞癌細(xì)胞凋亡水平的影響，利用流式細(xì)胞術(shù)檢測(cè)轉(zhuǎn)染miR150-5p-NC與miR-150-5p mimics的肝細(xì)胞癌HepG2細(xì)胞和SMMC-7221細(xì)胞的細(xì)胞凋亡的差異。結(jié)果顯示，在HepG2細(xì)胞中，miR-150-5p NC組細(xì)胞凋亡率為（2.146±0.327），miR-150-5p mimics組細(xì)胞凋亡率為（7.335±1.714），經(jīng)t檢驗(yàn)，差異有統(tǒng)計(jì)學(xué)意義（t= 10.332，P=0.012）。在SMMC-7221細(xì)胞中，miR-150-5p NC組細(xì)胞凋亡率為（3.159±0.273）%，miR-150-5p mimics組細(xì)胞凋亡率為（14.719±3.154）%，經(jīng)t檢驗(yàn)，差異有統(tǒng)計(jì)學(xué)意義（t=11.415，P=0.009）。與miR-150-5pNC組比較，miR-150-5p mimics組的肝細(xì)胞癌細(xì)胞的細(xì)胞凋亡率升高。以上結(jié)果提示，上調(diào)肝細(xì)胞癌細(xì)胞中miR-150-5p的表達(dá)水平可促進(jìn)肝細(xì)胞癌細(xì)胞的凋亡，進(jìn)一步證實(shí)miR-150-5p是潛在的腫瘤抑制分子。見(jiàn)圖4。

圖3 miR-150-5p對(duì)肝細(xì)胞癌細(xì)胞增殖的影響

2.5 miR-150-5對(duì)肝細(xì)胞癌細(xì)胞周期的影響

圖4 miR-150-5p對(duì)肝細(xì)胞癌細(xì)胞凋亡的影響

為研究miR-150-5p對(duì)肝細(xì)胞癌細(xì)胞周期的影響，利用流式細(xì)胞術(shù)檢測(cè)分別轉(zhuǎn)染miR-150-5p NC與miR-150-5p的肝細(xì)胞癌HepG2細(xì)胞和SMMC-7221細(xì)胞的細(xì)胞周期的差異。結(jié)果顯示，在HepG2細(xì)胞中，miR-150-5p NC組細(xì)胞合成期（synthesis phase，S期）細(xì)胞比率為（21.324±4.718）%，miR-150-5p mimics組S期細(xì)胞比率為（11.674±4.120）%，經(jīng)t檢驗(yàn)，差異有統(tǒng)計(jì)學(xué)意義（t=8.416，P=0.021）。在SMMC-7221細(xì)胞中，miR-150-5p NC組S期細(xì)胞比率為（29.728±5.723）%，miR-150-5p mimics組S期細(xì)胞比率為（18.344±4.115）%，經(jīng)t檢驗(yàn)，差異有統(tǒng)計(jì)學(xué)意義（t=7.195，P=0.034）。與miR-150-5p NC組比較，miR-150-5pmimcs組的肝細(xì)胞癌細(xì)胞的S期細(xì)胞的細(xì)胞比率降低，細(xì)胞周期被阻滯于第一間隙期（gap phase 1，G1期）。以上結(jié)果提示，上調(diào)肝細(xì)胞癌細(xì)胞中miR-150-5p的表達(dá)，可是細(xì)胞周期阻滯于G1期，增殖期S其細(xì)胞數(shù)目減少，細(xì)胞增殖收到抑制。見(jiàn)圖5。

2.6 miR-150-5p在肝細(xì)胞癌細(xì)胞系中靶向抑制ZNF609

圖5 miR-150-5p對(duì)肝細(xì)胞癌細(xì)胞周期的影響

為進(jìn)一步研究miR-150-5p在肝細(xì)胞癌中發(fā)揮抑癌作用的機(jī)制，綜合分析Target Scan 6.2和miRBse數(shù)據(jù)庫(kù)中miR-150-5p的潛在靶分子。生物信息學(xué)提示鋅指蛋白609（Zinc finger protein 609，ZNF609）為miR-150-5p新的潛在靶分子。Luciferase結(jié)果顯示，在肝細(xì)胞癌細(xì)胞系中ZNF609為miR-150-5p直接靶基因（P=0.031）（見(jiàn)圖6A）。進(jìn)一步實(shí)驗(yàn)結(jié)果顯示，轉(zhuǎn)染miR-150-5p-mimics的肝細(xì)胞癌HepG2細(xì)胞和SMMC-7721中ZNF609的mRNA表達(dá)水平降低（P=0.015）（見(jiàn)圖6B），以上結(jié)果提示，ZNF609為miR-150-5p在肝細(xì)胞癌細(xì)胞中的直接靶分子，miR-150-5p可直接抑制肝細(xì)胞癌細(xì)胞中ZNF609的表達(dá)水平。因此，miR-150-5p在肝細(xì)胞癌中發(fā)揮腫瘤抑制分子作用的潛在機(jī)制為直接靶向抑制ZNF609的表達(dá)。

圖6 miR-150-5P在肝細(xì)胞癌細(xì)胞系中靶向抑制ZNF609的表達(dá)

3 討論

既往研究發(fā)現(xiàn)，miR-150-5p在多種腫瘤中異常表達(dá)并參與調(diào)控腫瘤細(xì)胞的惡性表型。研究發(fā)現(xiàn)，miR-150在結(jié)腸癌組織中表達(dá)下調(diào)，miR-150-5p低表達(dá)的結(jié)腸癌患者的總體生存率縮短，對(duì)化療的反應(yīng)較miR-150高表達(dá)組也下降[12]。此外，miR-150可通過(guò)靶向抑制MUC4的表達(dá)而抑制結(jié)腸癌細(xì)胞的侵襲和轉(zhuǎn)移[13]。在食管鱗狀細(xì)胞癌中組織中，miR-150表達(dá)較正常食管黏膜下降[14]。miR-150可通過(guò)抑制ZEB1的表達(dá)而抑制食管鱗狀細(xì)胞癌細(xì)胞和卵巢癌細(xì)胞的侵襲能力[14-15]。相反，也有研究發(fā)現(xiàn)，miR-150在肺癌中表達(dá)上調(diào)，并可通過(guò)靶向抑制SRC激酶信號(hào)通知抑制分子1的表達(dá)而促進(jìn)肺癌細(xì)胞的增殖和轉(zhuǎn)移[16]。關(guān)于miR-150-5p在肝細(xì)胞癌中的表達(dá)及功能的研究尚不多，研究發(fā)現(xiàn)miR-150-5p在肝細(xì)胞癌組織中表達(dá)下調(diào)[17]，并可通過(guò)靶向調(diào)控MMP14分子而抑制肝細(xì)胞癌細(xì)胞的侵襲和轉(zhuǎn)移能力[11]。但目前尚無(wú)研究關(guān)注，miR-150-5p對(duì)肝細(xì)胞癌細(xì)胞增殖、凋亡和周期的影響。

本研究采用qRT-PCR法檢測(cè)人正常肝細(xì)胞系和肝細(xì)胞癌細(xì)胞系中miR-150-5p的表達(dá)差異，并利用 miR-150-5p mimics提高肝細(xì)胞細(xì)胞系中miR-150-5p的表達(dá)，而后采用MTT和流式細(xì)胞術(shù)等方法檢測(cè)miR-150-5p對(duì)肝細(xì)胞癌細(xì)胞增殖、凋亡和周期的影響。與既往文獻(xiàn)相符，研究發(fā)現(xiàn)，miR-150-5p在肝細(xì)胞癌細(xì)胞系HepG2和SMMC-7221中的表達(dá)較人正常肝細(xì)胞細(xì)胞系QZG和HL-7702中的表達(dá)降低。上調(diào)肝細(xì)胞癌細(xì)胞中miR-150-5p的表達(dá)后，肝細(xì)胞癌系細(xì)胞的增殖受到抑制，而細(xì)胞凋亡比率則升高。此外，細(xì)胞周期也被阻滯于G1期。進(jìn)一步的機(jī)制研究發(fā)現(xiàn)，miR-150-5p在肝細(xì)胞癌中發(fā)揮腫瘤抑制分子作用的潛在機(jī)制為直接靶向抑制ZNF609的表達(dá)。

多項(xiàng)研究表明，miRNA在肝細(xì)胞癌的發(fā)生及進(jìn)展中扮演著重要的角色。例如，miR-195在肝細(xì)胞癌組織中表達(dá)下調(diào)，并與肝細(xì)胞癌的血管生成、遠(yuǎn)處轉(zhuǎn)移和無(wú)病生存期呈負(fù)相關(guān)[18]。MiR-195發(fā)揮抑癌作用的潛在機(jī)制為其通過(guò)靶向抑制血管內(nèi)皮生長(zhǎng)因子（VEGF）和促轉(zhuǎn)移因子VAV2和CDC42的表達(dá)而抑制肝細(xì)胞癌細(xì)胞的侵襲和轉(zhuǎn)移[18]。MiR-140-5p在肝細(xì)胞癌中同樣表達(dá)下調(diào)，其表達(dá)水平與肝細(xì)胞癌多發(fā)結(jié)節(jié)、靜脈侵犯程度、囊形成、細(xì)胞核分化水平和總體積無(wú)病生存期密切相關(guān)[19]。本研究證實(shí)，miR-150-5p在肝細(xì)胞癌細(xì)胞中表達(dá)下調(diào)并在肝細(xì)胞癌中發(fā)揮腫瘤抑制分子的作用，其可通過(guò)調(diào)控ZNF609的表達(dá)水平而參與調(diào)控肝細(xì)胞癌細(xì)胞增殖、凋亡和周期。

綜上所述，本研究發(fā)現(xiàn)miR-150-5p在肝細(xì)胞癌細(xì)胞系中的表達(dá)較人正常肝細(xì)胞系下降，提高miR-150-5p在肝細(xì)胞癌細(xì)胞中的表達(dá)，可抑制肝細(xì)胞癌細(xì)胞的增殖能力，并促進(jìn)腫瘤細(xì)胞的凋亡率。此外，與NC組比較，miR-150-5p mimics組細(xì)胞周期中S期細(xì)胞所占比例也降低。進(jìn)一步的研究顯示，miR-150-5p在肝細(xì)胞癌細(xì)胞中發(fā)揮抑癌分子作用的潛在機(jī)制為其靶向抑制ZNF609的表達(dá)。本研究進(jìn)一步揭示，miR-150-5p在肝細(xì)胞癌中的表達(dá)及其對(duì)肝細(xì)胞癌細(xì)胞增殖、凋亡和周期的作用，為肝細(xì)胞癌的治療提供潛在的分子靶點(diǎn)。

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（張西倩 編輯）

Effects of miR-150-5p on proliferation,apoptosis and cell cycle of hepatocellular carcinoma cells

Yue-qiu Zhao1,Zheng-bao Zhang1,Hong-xia Guo1,Chuan-fei Zhan2,Hao Lu2,Xiao-feng Qian2
(1.Nanjing Health School,Jiangsu Union Technical Institute,Nanjing,Jiangsu 210038,China;2. Department of Liver Surgery,Jiangsu Provincial Hospital,Nanjing,Jiangsu 210029,China)

Objective To investigate the expression and effect of miR-150-5p on cell proliferation,cell apoptosis and cell cycle in human hepatocellular carcinoma cells.Methods The expression of miR-150-5p in hepatocellular carcinoma cells and normal human liver cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR).Proliferation of hepatocellular carcinoma cells was analyzed by MTT.Cell apoptosis and cell cycle of hepatocellular carcinoma cells were analyzed by flow cytometry.Luciferase assay was performed to detect whether ZNF609 was the direct target of miR-150-5p in the hepatocellular carcinoma cells.Results The expression of miR-150-5p was significantly down regulated in the hepatocellular carcinoma cells compared to the normal human liver cells.Proliferation of the hepatocellular carcinoma cells was significantly inhibited by miR-150-5p,but cell apoptosis was significantly promoted.Cell cycle of the hepat ocellular carcinoma cells was arrested at G1 phage after transfection of miR-150-5p mimics.Further investigation proved ZNF609 was the direct target of miR-150-5p in hepatocellular carcinoma cells.Conclusions miR-150-5p may function as tumor suppressor and play important roles in cell proliferation,cell apoptosis and cell cycle in hepatocellular carcinoma cells by directly targeting ZNF609.

miR-150-5p;cell proliferation;cell apoptosis;cell cycle;ZNF609

臨床研究·論著

R735

A

10.3969/j.issn.1005-8982.2017.04.006

1005-8982（2017）04-0027-06

2016-07-13