姜黃素通過(guò)下調(diào)nectin-4抑制肺癌PC-9細(xì)胞的遷移和侵襲*

陳 君， 焦德敏， 唐夏莉， 王 劍， 李 優(yōu)， 陳清勇

(中國(guó)人民解放軍第一一七醫(yī)院呼吸內(nèi)科，浙江 杭州 310004)

姜黃素通過(guò)下調(diào)nectin-4抑制肺癌PC-9細(xì)胞的遷移和侵襲*

陳 君， 焦德敏， 唐夏莉， 王 劍， 李 優(yōu)， 陳清勇△

(中國(guó)人民解放軍第一一七醫(yī)院呼吸內(nèi)科，浙江 杭州 310004)

目的： 探討姜黃素對(duì)肺癌PC-9細(xì)胞遷移和侵襲的抑制作用及其與nectin-4表達(dá)的關(guān)系。方法: 用MTT、劃痕修復(fù)和Transwell小室實(shí)驗(yàn)檢測(cè)姜黃素對(duì)肺癌PC-9細(xì)胞活力、遷移和侵襲能力的影響；用Western blot技術(shù)檢測(cè)姜黃素對(duì)nectin-4表達(dá)及AKT通路的影響；經(jīng)siRNA干擾nectin-4后觀察姜黃素對(duì)PC-9細(xì)胞活力、遷移、侵襲及AKT通路的影響。結(jié)果:姜黃素能抑制PC-9細(xì)胞活力，10、20 μmol/L姜黃素組與對(duì)照組相比,劃痕修復(fù)率降低，穿膜細(xì)胞數(shù)目顯著下降。姜黃素作用肺癌PC-9細(xì)胞24 h后,nectin-4表達(dá)下調(diào)。轉(zhuǎn)染siNectin-4 48 h和72 h后，siNectin-4組比對(duì)照組的細(xì)胞活力顯著降低(P<0.01)，劃痕修復(fù)率及穿膜細(xì)胞數(shù)目顯著下降(P<0.01)。姜黃素與siNectin-4均抑制了PC-9細(xì)胞AKT通路的激活。姜黃素與siNectin-4聯(lián)用時(shí)，細(xì)胞活力降低(P<0.01)，劃痕修復(fù)率、細(xì)胞侵襲能力及AKT磷酸化水平下降。結(jié)論: 在肺癌PC-9細(xì)胞中，姜黃素通過(guò)下調(diào)nectin-4表達(dá)、調(diào)控下游AKT通路來(lái)抑制細(xì)胞遷移和侵襲。

姜黃素； 肺癌； 細(xì)胞遷移； 細(xì)胞侵襲； Nectin-4

肺癌的發(fā)病率和死亡率居于惡性腫瘤的首位，近50年來(lái)肺癌的發(fā)生率和死亡率均明顯升高。肺癌轉(zhuǎn)移是引起肺癌患者死亡的主要原因，如何抑制肺癌轉(zhuǎn)移是目前肺癌治療需要解決的重要問(wèn)題。姜黃素(curcumin, Cur)是一種姜黃根莖中分離提取出來(lái)的天然小分子多酚化合物，具有廣泛的藥理作用。近年來(lái)，大量文獻(xiàn)報(bào)道姜黃素對(duì)多種癌細(xì)胞均有抑制其增殖、遷移和侵襲的作用[1-4]。我們以往的研究也發(fā)現(xiàn)姜黃素可調(diào)節(jié)抑制Rho家族的Cdc42和Rac通路影響肺癌細(xì)胞骨架重排，抑制肺癌細(xì)胞的侵襲和遷移[5-6]。Nectin家族屬于免疫球蛋白細(xì)胞黏附分子，nectins和afadin組成的細(xì)胞間黏附系統(tǒng)在腫瘤細(xì)胞的粘附和侵襲中發(fā)揮重要作用。近年來(lái)的研究發(fā)現(xiàn)外源性nectin-4的表達(dá)將通過(guò)激活Rac通路提高COS-7 和 NIH-3T3細(xì)胞的絲狀偽足形成率和侵襲能力，且非小細(xì)胞肺癌患者血清中的nectin-4含量比健康人高，nectin-4可作為肺癌的發(fā)展轉(zhuǎn)移過(guò)程中新型的血清學(xué)和組織學(xué)標(biāo)志物[7]。但是nectin-4是否參與姜黃素對(duì)肺癌轉(zhuǎn)移的抑制過(guò)程尚不清楚。

本研究以肺癌PC-9 細(xì)胞為研究對(duì)象，通過(guò)不同濃度姜黃素處理和nectin-4 RNA干擾技術(shù)來(lái)檢測(cè)兩者抑制肺癌PC-9 細(xì)胞的遷移和侵襲的能力，并從中尋找兩者的關(guān)系。

材 料 和 方 法

1 材料

肺腺癌細(xì)胞株P(guān)C-9購(gòu)自ATCC細(xì)胞庫(kù)；姜黃素、MTT和二甲基亞砜(DMSO)均購(gòu)自Sigma；小牛血清購(gòu)自Gibco；1640培養(yǎng)基和牛血清白蛋白購(gòu)自杭州吉諾公司；轉(zhuǎn)染試劑采用Lipofectamine 2000購(gòu)自Invitrogen；BCA試劑盒購(gòu)自凱基生物；抗nectin-4抗體購(gòu)自R&D；抗磷酸化及總AKT抗體購(gòu)自CST；抗GAPDH抗體購(gòu)自Abcam；HRP標(biāo)記的II抗購(gòu)自Jackson ImmunoResearch；ECL化學(xué)發(fā)光試劑盒購(gòu)自Pierce。蛋白質(zhì)電泳和轉(zhuǎn)膜設(shè)備和酶標(biāo)儀購(gòu)自Bio-Rad。

2 方法

2.1 細(xì)胞培養(yǎng)和轉(zhuǎn)染 細(xì)胞在含有10%小牛血清的RPMI-1640培養(yǎng)基中置于37 ℃、5% CO2培養(yǎng)箱中培養(yǎng)。采用0.25% 胰酶+0.02% EDTA溶液消化細(xì)胞。選用生長(zhǎng)情況良好的對(duì)數(shù)生長(zhǎng)期細(xì)胞進(jìn)行實(shí)驗(yàn)。Nectin-4 siRNA (siNectin-4)由上海吉瑪公司合成，經(jīng)前期實(shí)驗(yàn)[8]，選用干擾效果最佳序列，上游序列為5’-CAGCAGAUGACCCAGAAAUTT-3’，下游序列為3’-AUUUCUGGGUCAUCUGCUGTT-5’。

2.2 MTT實(shí)驗(yàn)測(cè)試細(xì)胞活力 將對(duì)數(shù)生長(zhǎng)期細(xì)胞經(jīng)胰酶消化后配成4×107/L濃度的細(xì)胞懸液，每孔100 μL接入96孔板。細(xì)胞24 h貼壁后，進(jìn)行實(shí)驗(yàn)，復(fù)孔5個(gè)。檢測(cè)時(shí)，每孔換液加入100 μL 5 g/L MTT，37 ℃孵育4 h，棄上清，每孔加入150 μL DMSO，振蕩使沉淀充分溶解，酶標(biāo)儀490 nm檢測(cè)吸光度(A)值。

2.3 Western blot實(shí)驗(yàn) 處理完成的細(xì)胞用裂解液提取蛋白，使用BCA試劑盒按說(shuō)明書(shū)測(cè)試蛋白濃度并配制樣品，上樣20 μg。SDS-PAGE及轉(zhuǎn)膜，用含5%牛血清白蛋白的TBST封閉目的蛋白，常溫封閉2 h，TBST洗膜。 I 抗按抗體說(shuō)明書(shū)使用碧云天 I 抗稀釋液稀釋后4 ℃孵育過(guò)夜，TBST洗膜5 min×3次， II 抗用封閉液稀釋合適比例后37 ℃孵育2 h，TBST洗膜3次后用ECL法對(duì)目的蛋白進(jìn)行發(fā)光，在凝膠成像系統(tǒng)中拍照，GAPDH作為內(nèi)參照。用ImageJ軟件測(cè)試目的蛋白條帶的灰度值進(jìn)行分析。

2.4 劃痕修復(fù)實(shí)驗(yàn) 處理過(guò)的細(xì)胞在12孔板中培養(yǎng)，第2天用10 μL槍頭沿直線在孔底劃線，PBS清洗2次，加入含相應(yīng)姜黃素濃度或不含姜黃素的2.5%血清的新鮮培養(yǎng)基培養(yǎng)，在倒置顯微鏡下拍照(×100)，在劃痕邊緣等距量取距離，培養(yǎng)24 h后，再拍照同樣位置量取距離。劃痕修復(fù)率(%)=(0 h劃痕寬度-24 h劃痕寬度)/0 h劃痕寬度×100%。

2.5 Transwell小室實(shí)驗(yàn) 加藥或轉(zhuǎn)染后的細(xì)胞用無(wú)血清培養(yǎng)基消化配成1.5×108/L的細(xì)胞懸液，鋪好Matrigel的上室中每孔加入200 μL細(xì)胞懸液，下室加入700 μL含有相應(yīng)濃度藥物的1640培養(yǎng)基(10%血清)。培養(yǎng)箱內(nèi)培育12 h后，PBS清洗小室2次，冰甲醇固定細(xì)胞20 min后風(fēng)干，1‰ 結(jié)晶紫染色30 min，清水清洗小室3次，棉簽輕輕吸干上室水分，倒置顯微鏡下拍照(×100)。用ImageJ計(jì)算細(xì)胞數(shù)目。

3 統(tǒng)計(jì)學(xué)處理

所有實(shí)驗(yàn)均重復(fù)3 次，實(shí)驗(yàn)數(shù)據(jù)用均數(shù)±標(biāo)準(zhǔn)差(mean±SD)表示，采用SPSS Statistics 22進(jìn)行數(shù)據(jù)分析。使用GraphPad Prism 5制圖。多組間比較應(yīng)用單因素方差分析，組間兩兩比較應(yīng)用Bonferroni 檢驗(yàn)，以P<0.05 為差異有統(tǒng)計(jì)學(xué)意義。

結(jié) 果

1 姜黃素抑制PC-9細(xì)胞活力、遷移和侵襲

姜黃素處理PC-9細(xì)胞72 h后，MTT實(shí)驗(yàn)結(jié)果顯示10～40 μmol/L姜黃素均能抑制PC-9細(xì)胞的生長(zhǎng)，抑制率分別是(23.93±1.82)%、(58.97±3.21)%、(76.19±5.61)%和(76.44+3.47)%，隨著給藥劑量的增加，姜黃素對(duì)肺癌細(xì)胞的生長(zhǎng)抑制作用顯著增強(qiáng)(P<0.01)，見(jiàn)圖1A。劃痕修復(fù)實(shí)驗(yàn)結(jié)果顯示，與對(duì)照組相比，姜黃素10 μmol/L和20 μmol/L組劃痕愈合率分別下降了(13.32±4.40)%和(47.22±2.71)%(P<0.05)，表明姜黃素能抑制PC-9細(xì)胞的遷移，見(jiàn)圖1B。Transwell小室實(shí)驗(yàn)結(jié)果顯示，對(duì)照組、姜黃素10 μmol/L組和姜黃素20 μmol/L組侵襲至Transwell小室濾膜下表面的細(xì)胞數(shù)分別為110±6、69±3和51±4，姜黃素10 μmol/L和20 μmol/L組較對(duì)照組細(xì)胞穿膜數(shù)目顯著下降(P<0.01)，表明姜黃素能抑制PC-9細(xì)胞的侵襲，見(jiàn)圖1C。

Figure 1. The growth inhibition rate, migration and invasion of PC-9 cells treated with curcumin (Cur). A: the growth inhibition rate of PC-9 cells after curcumin treatment; B: the migration of PC-9 cells after curcumin 10 μmol/L and 20 μmol/L treatment; C: the invasion of PC-9 cells after curcumin 10 μmol/L and 20 μmol/L treatment. Mean±SD.n=3.*P<0.05,**P<0.01vscontrol.

圖1 姜黃素對(duì)PC-9細(xì)胞活力、遷移和侵襲的作用

2 姜黃素下調(diào)nectin-4蛋白的表達(dá)

Western blot檢測(cè)結(jié)果顯示，10、20、30和40 μmol/L姜黃素作用肺癌PC-9細(xì)胞24 h后，nectin-4蛋白的表達(dá)均顯著下降(P<0.01)，見(jiàn)圖2。

3 干擾nectin-4抑制PC-9細(xì)胞活力、遷移和侵襲

通過(guò)前期實(shí)驗(yàn)[8]，我們選取干擾效果最佳的基因片段，構(gòu)建siNectin-4。Western blot檢測(cè)結(jié)果顯示,與對(duì)照組相比，siNectin-4組的nectin-4蛋白表達(dá)明顯下降(P<0.01),見(jiàn)圖3A。MTT實(shí)驗(yàn)結(jié)果顯示siNectin-4分別轉(zhuǎn)染PC-9細(xì)胞6 h、24 h、48 h和72 h后，細(xì)胞活力與對(duì)照組相比分別下降了(2.98±5.11)%、(4.59±11.46)%、(29.31±9.73)%和(90.66±19.02)%，轉(zhuǎn)染48 h和72 h后，siNectin-4組與對(duì)照組相比差異有統(tǒng)計(jì)學(xué)意義(P<0.01)，表明干擾nectin-4能抑制PC-9細(xì)胞活力，見(jiàn)圖3B。劃痕修復(fù)實(shí)驗(yàn)結(jié)果顯示,與對(duì)照組相比，siNectin-4組的劃痕愈合率下降了(28.76±5.66)%，差異具有統(tǒng)計(jì)學(xué)意義(P<0.01)，表明siNectin-4能抑制PC-9細(xì)胞的遷移，見(jiàn)圖3C。Transwell小室實(shí)驗(yàn)結(jié)果顯示,與對(duì)照組相比，siNectin-4組侵襲至Transwell小室濾膜下表面的細(xì)胞數(shù)顯著下降(P<0.01)，表明siNectin-4能抑制PC-9細(xì)胞的侵襲，見(jiàn)圖3D。

Figure 2. The protein levels of nectin-4 in the PC-9 cells after curcumin (Cur) treatment. Mean±SD.n=3.**P<0.01vscontrol.

圖2 姜黃素作用對(duì)PC-9細(xì)胞中nectin-4蛋白表達(dá)的影響

4 姜黃素和干擾nectin-4均能抑制AKT通路

10～40 μmol/L濃度姜黃素均能抑制PC-9細(xì)胞AKT的磷酸化(P<0.01)，siNectin-4也抑制了PC-9細(xì)胞的AKT磷酸化(P<0.01)，總AKT表達(dá)無(wú)明顯差異，見(jiàn)圖4。

5 姜黃素與siNectin-4聯(lián)合對(duì)PC-9細(xì)胞活力、遷移、侵襲及AKT通路的影響

siNectin-4與姜黃素合用后，細(xì)胞活力、劃痕修復(fù)率、細(xì)胞遷移及侵襲能力均較單用姜黃素組下降(P<0.01)。同時(shí)檢測(cè)它們AKT通路磷酸化的變化，發(fā)現(xiàn)與姜黃素單用組比較，siNectin-4與姜黃素合用明顯抑制AKT通路磷酸化，見(jiàn)圖5。

Figure 3. The viability, migration and invasion of the PC-9 cells transfected with siNectin-4. A: the protein levels of nectin-4 in the PC-9 cells transfected with siNectin-4; B: the viability of the PC-9 cells transfected with siNectin-4; C: the migration of PC-9 cells transfected with siNectin-4; D: the invasion of the PC-9 cells transfected with siNectin-4. Mean±SD.n=3.**P<0.01vsNC.

圖3 干擾nectin-4的表達(dá)對(duì)PC-9細(xì)胞活力、遷移和侵襲作用的影響

Figure 4.The phosphorylation level of AKT in the PC-9 cells treated with curcumin (Cur) or transfected with siNectin-4. A: the protein levels of p-AKT in the PC-9 cells after curcumin treatment; B: the protein levels of p-AKT in the PC-9 cells transfected with siNectin-4. Mean±SD.n=3.**P<0.01vscontrol;##P<0.01vsNC.

圖4 姜黃素與siNectin-4分別對(duì)PC-9細(xì)胞AKT磷酸化水平的影響

討 論

姜黃素具有抗氧化、抗衰老及抗腫瘤的作用。以往的研究從多個(gè)方面闡釋了姜黃素的抗腫瘤機(jī)制[9]，例如在肺癌H460、A427和A549細(xì)胞中，姜黃素通過(guò)激活p53上調(diào)miR-192-5p和miR-215表達(dá)，下調(diào)靶基因XIAP，從而促進(jìn)細(xì)胞凋亡[10]；姜黃素能通過(guò)抑制Rac1/PAK1信號(hào)通路降低MMP-2和MMP-9表達(dá)，抑制肺癌801D細(xì)胞的遷移和侵襲[5]；姜黃素還可通過(guò)抑制AKT/mTOR/p70S6K通路誘導(dǎo)自噬和抑制細(xì)胞毒性[11]。在乳腺癌MDA-MB-231細(xì)胞中，Soung等[12]發(fā)現(xiàn)姜黃素抑制了由EGF引起的整合素α6β4位置的變化，從而抑制了細(xì)胞的增殖和絲狀偽足形成。本實(shí)驗(yàn)結(jié)果表明，姜黃素能下調(diào)細(xì)胞黏附分子nectin-4表達(dá)并通過(guò)AKT途徑抑制肺癌PC-9細(xì)胞的活力、遷移和侵襲。

Nectins是Ca2+依賴的免疫球蛋白樣細(xì)胞間黏附分子，該家族成員通過(guò)afadin和細(xì)胞骨架聯(lián)系，調(diào)控細(xì)胞運(yùn)動(dòng)[13]。Nectin-4是該家族研究較為清楚的一個(gè)分子，nectin-4與nectin-1可通過(guò)V域相互傳動(dòng)作用[14]。Honda等[15]報(bào)道nectins通過(guò)激活Cdc42和Rac使JNK通路活化。 Kanzaki等[16]也發(fā)現(xiàn)nectins可以調(diào)控血小板源性生長(zhǎng)因子受體(platelet-derived growth factor receptor,PDGFR)及其下游通路，影響成纖維細(xì)胞的生長(zhǎng)過(guò)程。我們前期實(shí)驗(yàn)[8]表明，降低nectin-4的表達(dá)將抑制肺癌A549和PC-9細(xì)胞的遷移和侵襲能力，說(shuō)明nectin-4在肺癌細(xì)胞生長(zhǎng)中起到關(guān)鍵作用，nectin-4可成為預(yù)測(cè)肺癌轉(zhuǎn)移的生物標(biāo)志物。在本研究中，我們發(fā)現(xiàn)nectin-4 參與PC-9細(xì)胞的生長(zhǎng)和遷移過(guò)程。因此，nectins作為調(diào)控細(xì)胞黏附的重要分子，可能是腫瘤細(xì)胞的遷移和侵襲的重要靶點(diǎn)。

為進(jìn)一步明確姜黃素與nectin-4之間的關(guān)系，還需要通過(guò)動(dòng)物水平來(lái)研究nectin-4抑制腫瘤的功能。目前，Pavlova等[17]已證實(shí)干擾nectin-4能抑制小鼠體內(nèi)腫瘤生長(zhǎng)，乳腺癌SUM190細(xì)胞中nectin-4促進(jìn)了不貼壁細(xì)胞中的細(xì)胞間接觸組裝，與整合素 β4相互作用并持續(xù)激活了Src家族激酶。Dipon等[18]則進(jìn)一步研究nectin-4與5-FU耐藥的關(guān)系，發(fā)現(xiàn)nectin-4通過(guò)PI3K-AKT通路增強(qiáng)結(jié)腸癌細(xì)胞對(duì)5-FU的耐藥性，而該通路與肺癌細(xì)胞的增殖、遷移和侵襲緊密相關(guān)[19]。與該機(jī)制類似，我們通過(guò)Western blot實(shí)驗(yàn)發(fā)現(xiàn)姜黃素抑制了nectin-4蛋白的表達(dá)，且同時(shí)抑制了肺癌PC-9細(xì)胞的活力、遷移、侵襲及AKT通路的磷酸化，與干擾nectin-4后的表現(xiàn)一致。進(jìn)一步我們發(fā)現(xiàn)姜黃素與干擾nectin-4聯(lián)用時(shí)，細(xì)胞活力、遷移、侵襲能力及AKT磷酸化水平更低，這些結(jié)果說(shuō)明姜黃素抑制肺癌PC-9細(xì)胞生長(zhǎng)、遷移的細(xì)胞機(jī)制與下調(diào)nectin-4有關(guān)，姜黃素可能是通過(guò)下調(diào)nectin-4來(lái)抑制細(xì)胞活力、遷移、侵襲。

不過(guò)，本研究發(fā)現(xiàn)在肺癌PC-9細(xì)胞中，干擾nectin-4不僅能抑制細(xì)胞活力，而且可以抑制細(xì)胞遷移與侵襲，這個(gè)結(jié)果與Pavlova等[17]提出在乳腺癌SUM190細(xì)胞中nectin-4未引起EMT變化和轉(zhuǎn)移變化的結(jié)果有所不同，我們認(rèn)為不同的結(jié)果可能與不同細(xì)胞有關(guān)。

Figure 5. The effects of curcumin (Cur) treatment and transfection with siNectin-4 on the viability, migration, invasion and AKT phosphorylation in the PC-9 cells. A: the viability of the PC-9 cells transfected with siNectin-4 and treated with curcumin; B: the migration of the PC-9 cells transfected with siNectin-4 and treated with curcumin; C: the invasion of the PC-9 cells transfected with siNectin-4 and treated with curcumin; D: the protein levels of p-AKT in the PC-9 cells transfected with siNectin-4 and treated with curcumin. Mean±SD.n=3.**P<0.01vsNC+Cur 10 μmol/L;##P<0.01vsNC+Cur 20 μmol/L.

圖5 姜黃素與干擾Nectin-4聯(lián)合應(yīng)用對(duì)PC-9細(xì)胞的活力、遷移、侵襲及AKT磷酸化水平的影響

總之，本研究發(fā)現(xiàn)了姜黃素與干擾nectin-4具有同樣的抑制肺癌PC-9細(xì)胞活力、遷移、侵襲和AKT磷酸化的能力，姜黃素能降低nectin-4蛋白的表達(dá)，姜黃素與干擾nectin-4聯(lián)合應(yīng)用使細(xì)胞活力、遷移、侵襲能力及AKT磷酸化水平更低，推測(cè)姜黃素抑制細(xì)胞活力、遷移、侵襲與下調(diào)nectin-4有關(guān)，nectin-4可能是姜黃素抑制腫瘤的途徑之一。研究結(jié)果為姜黃素抑制腫瘤細(xì)胞生長(zhǎng)的機(jī)制提供了新思路，為nectin-4作為新的肺癌轉(zhuǎn)移治療的靶向分子提供了實(shí)驗(yàn)證據(jù)。

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(責(zé)任編輯： 陳妙玲， 羅 森)

Curcumin inhibits migration and invasion of lung cancer PC-9 cells via down-regulation of nectin-4 expression

CHEN Jun, JIAO De-min, TANG Xia-li, WANG Jian, LI You, CHEN Qing-yong

(DepartmentofRespiratoryMedicine,The117thHospitalofPLA,Hangzhou310004,China.E-mail:cqyong117@163.com)

AIM: To investigate the effects of curcumin on the abilities of migration and invasion in the lung cancer PC-9 cells, and to observe the relationship between curcumin and nectin-4 expression. METHODS: The viability, migration and invasion of lung cancer PC-9 cells treated with curcumin or transfected with siNectin-4 were measured by MTT assay, wound healing test and Transwell assay, respectively. The protein levels of nectin-4, p-AKT and AKT in the PC-9 cells treated with curcumin or transfected with siNectin-4 were detected by Western blot. RESULTS: Curcumin inhibited the viability of PC-9 cells. The wound healing rates and the numbers of the transmembrane cells in curcumin 10 μmol/L and 20 μmol/L groups were decreased compared with control group without curcumin treatment. The expression level of nectin-4 was reduced after curcumin treatment for 24 h. The viability of the PC-9 cells was significantly inhibited after transfected with siNectin-4 for 48 h or 72 h (P<0.01), and the wound healing rates was decreased in siNectin-4 group compared with NC group (P<0.01). The numbers of the transmembrane cells in siNectin-4 group was significantly reduced (P<0.01). Curcumin and knockdown of nectin-4 suppressed the activation of AKT pathway in PC-9 cells. In siNectin-4+curcumin group, the cell viability reduced compared with curcumin group, and wound healing rates, cell invasive ability and AKT phosphorylation levels were decreased. CONCLUSION: Curcumin inhibits migration and invasion of the lung cancer PC-9 cells via down-regulation of nectin-4 expression and inhibition of AKT pathway.

Curcumin; Lung cancer; Cell migration; Cell invasion; Nectin-4

1000- 4718(2017)02- 0271- 07

2016- 08- 20

2016- 10- 19

浙江省醫(yī)藥衛(wèi)生科技計(jì)劃(No.2011KYA139)；浙江省科技廳公益性技術(shù)應(yīng)用研究計(jì)劃(No.2014C33277)；杭州市科技發(fā)展計(jì)劃(No.20130633B29; No.20140633B40)；浙江省自然科學(xué)基金資助項(xiàng)目(No.LY12H16001)

R730.23

A

10.3969/j.issn.1000- 4718.2017.02.013

雜志網(wǎng)址： http://www.cjpp.net

△通訊作者 Tel: 0571-28084872; E-mail: cqyong117@163.com