卡非佐米聯(lián)合米托坦對(duì)人腎上腺皮質(zhì)癌細(xì)胞H295R、SW13增殖及細(xì)胞周期的影響

韋慶臣，譚茹尹，楊海燕，黃振興，梁杏歡，秦映芬，羅佐杰

(廣西醫(yī)科大學(xué)第一附屬醫(yī)院，南寧530021)

卡非佐米聯(lián)合米托坦對(duì)人腎上腺皮質(zhì)癌細(xì)胞H295R、SW13增殖及細(xì)胞周期的影響

韋慶臣，譚茹尹，楊海燕，黃振興，梁杏歡，秦映芬，羅佐杰

(廣西醫(yī)科大學(xué)第一附屬醫(yī)院，南寧530021)

目的探討卡非佐米聯(lián)合米托坦對(duì)人腎上腺皮質(zhì)癌(ACC)細(xì)胞H295R、SW13增殖及細(xì)胞周期的影響。方法體外培養(yǎng)人ACC細(xì)胞H295R、SW13，采用MTT法確定米托坦作用H295R、SW13細(xì)胞48 h時(shí)的半數(shù)有效抑制濃度(IC50)值分別為18.42、62.37 μmol/L，卡非佐米分別為3.86、11.62 μmol/L。將兩種對(duì)數(shù)生長(zhǎng)期細(xì)胞隨機(jī)分為對(duì)照組、卡非佐米組、米托坦組及序貫給藥的卡非佐米→米托坦組、卡非佐米+米托坦組、米托坦→卡非佐米組，給藥濃度分別為兩種藥物IC50值的1/8、1/4、1/2、1、2倍，檢測(cè)各濃度干預(yù)48 h時(shí)各組細(xì)胞增殖抑制率。根據(jù)Chou-Talaly公式計(jì)算IC50及2倍IC50作用下兩種藥物序貫給藥時(shí)的聯(lián)合指數(shù)(CI)。將兩種對(duì)數(shù)生長(zhǎng)期細(xì)胞隨機(jī)分為對(duì)照組、米托坦單藥組(25、50 μmol/L)、卡非佐米單藥組(1 μmol/L)、卡非佐米(1 μmol/L)→米托坦(25、50 μmol/L)組，藥物干預(yù)48 h時(shí)檢測(cè)細(xì)胞增殖抑制率。將兩種對(duì)數(shù)生長(zhǎng)期細(xì)胞隨機(jī)分為空白組、米托坦組、卡非佐米組、卡非佐米→米托坦組，后三組以IC50藥物濃度干預(yù)96 h，采用流式細(xì)胞儀檢測(cè)細(xì)胞周期。結(jié)果H295R及SW13細(xì)胞在IC50及2倍IC50濃度下，卡非佐米→米托坦組細(xì)胞增殖抑制率均高于米托坦→卡非佐米組及卡非佐米+米托坦組，米托坦→卡非佐米組均高于卡非佐米+米托坦組，組間比較P均<0.05。H295R和SW13細(xì)胞在米托坦、卡非佐米IC50及2倍IC50濃度條件下，卡非佐米→米托坦組和米托坦+卡非佐米組中兩藥的CI值均<1，且卡非佐米→米托坦組兩藥的CI值均小于米托坦+卡非佐米組(P均<0.05)；米托坦→卡非佐米組中兩藥的CI值均<1，且均高于卡非佐米→米托坦組和米托坦+卡非佐米組(P均<0.05)。1 μmol/L卡非佐米與25、50 μmol/L米托坦聯(lián)合的卡非佐米→米托坦組H295R及SW13增殖抑制率分別高于25、50 μmol/L米托坦組(P均<0.05)。四組G1、G2/M、S期細(xì)胞百分比比較差異均無(wú)統(tǒng)計(jì)學(xué)意義(P均>0.05)。結(jié)論卡非佐米聯(lián)合米托坦對(duì)ACC細(xì)胞存在序貫依賴性的協(xié)同抑制細(xì)胞增殖作用，給予卡非佐米后給予米托坦的抗增殖作用最佳，但兩藥聯(lián)合對(duì)細(xì)胞周期無(wú)明顯影響。

腎上腺皮質(zhì)癌；卡非佐米；米托坦；細(xì)胞增殖；細(xì)胞周期

腎上腺皮質(zhì)癌(ACC)是來(lái)源于腎上腺皮質(zhì)的惡性腫瘤，臨床罕見(jiàn)，年發(fā)病率為0.7/100萬(wàn)～2.0/100萬(wàn)[1]。臨床對(duì)ACC患者多采用手術(shù)治療，術(shù)后給予米托坦單藥或聯(lián)合細(xì)胞毒藥物輔助治療[2]；但患者5年生存率只有16%～47%，已發(fā)生腫瘤轉(zhuǎn)移者5年生存率不足10%[3]。米托坦可特異性抑制腎上腺SOAT1酶活性，使游離膽固醇和游離脂肪酸在細(xì)胞內(nèi)堆積，誘發(fā)內(nèi)質(zhì)網(wǎng)應(yīng)激，導(dǎo)致細(xì)胞凋亡[5]?？ǚ亲裘资切乱淮叨冗x擇性不可逆蛋白酶體抑制劑，可阻斷細(xì)胞內(nèi)泛素—蛋白酶體蛋白降解途徑，導(dǎo)致大量泛素化蛋白在細(xì)胞內(nèi)堆積，引起細(xì)胞內(nèi)質(zhì)網(wǎng)應(yīng)激，激活未折疊蛋白反應(yīng)(UPR)，導(dǎo)致細(xì)胞死亡[6]。上述兩種藥物均主要通過(guò)內(nèi)質(zhì)網(wǎng)應(yīng)激來(lái)發(fā)揮抗腫瘤作用，兩者聯(lián)用可能存在協(xié)同抗腫瘤作用。2016年6～12月我們進(jìn)行本研究，探討卡非佐米聯(lián)合米托坦對(duì)人ACC細(xì)胞H295R、SW13增殖及細(xì)胞周期的影響。

1 材料

ACC細(xì)胞株SW13、H295R分別購(gòu)自中國(guó)科學(xué)院上海細(xì)胞庫(kù)及北納創(chuàng)聯(lián)生物技術(shù)研究院。卡非佐米和米托坦粉劑均購(gòu)自大連美侖生物技術(shù)有限公司，DMEM高糖培養(yǎng)基購(gòu)自美國(guó)Hyclone公司，MTT購(gòu)自上海碧云天生物技術(shù)研究所。細(xì)胞周期與細(xì)胞凋亡檢測(cè)試劑盒購(gòu)自BestBio公司。

2 方法與結(jié)果

2.2 細(xì)胞培養(yǎng) 將SW13及H295R細(xì)胞置于含有10% FBS的F12/DMEM 1∶1完全培養(yǎng)液中，于溫度37 ℃、濕度90%、5% CO2培養(yǎng)箱內(nèi)培養(yǎng)，定期觀察細(xì)胞形態(tài)及生長(zhǎng)狀況，取對(duì)數(shù)生長(zhǎng)期細(xì)胞進(jìn)行實(shí)驗(yàn)。

2.3 卡非佐米、米托坦對(duì)SW13及H295R細(xì)胞的半數(shù)有效抑制濃度(IC50)確定 取對(duì)數(shù)生長(zhǎng)期SW13及H295R細(xì)胞，分別以4×103個(gè)/孔接種于96孔板，培養(yǎng)12 h，待細(xì)胞完全貼壁生長(zhǎng)后以不同濃度的卡非佐米(0.5、1、2、4、8、16、32 μmol/L)或米托坦(2、4、8、16、32、64 μmol/L)分別處理，每個(gè)濃度設(shè)6個(gè)復(fù)孔。藥物干預(yù)48 h時(shí)，各孔加入MTT 20 μL，置于培養(yǎng)箱繼續(xù)培養(yǎng)4 h；小心吸去上清液，加入DMSO 200 μL，置于搖床振蕩混勻10 min；采用酶標(biāo)儀檢測(cè)各孔在570 nm波長(zhǎng)處的光密度(OD)值，計(jì)算細(xì)胞增殖抑制率。細(xì)胞增殖抑制率=(1-實(shí)驗(yàn)組OD值/對(duì)照組OD值)×100%。實(shí)驗(yàn)重復(fù)3次，取平均值。采用GraphPad Prism5.0軟件計(jì)算卡非佐米、米托坦分別對(duì)H295R及SW13細(xì)胞的IC50。結(jié)果顯示，米托坦對(duì)H295R和SW13細(xì)胞的IC50值分別為18.42、62.37 μmol/L，卡非佐米分別為3.86、11.62 μmol/L。

2.4 卡非佐米聯(lián)合米托坦對(duì)H295R和SW13細(xì)胞增殖的影響 取對(duì)數(shù)生長(zhǎng)期SW13及H295R細(xì)胞，分別以4×103個(gè)/孔接種于96孔板，隨機(jī)分為對(duì)照組、卡非佐米組、米托坦組及序貫給藥組卡非佐米聯(lián)合米托坦組(聯(lián)合組)。對(duì)照組正常培養(yǎng)液培養(yǎng)96 h?？ǚ亲裘捉M：先給予卡非佐米干預(yù)48 h，然后將培養(yǎng)液吸出，加入少量PBS將卡非佐米洗去，加入不含藥物的完全培養(yǎng)液繼續(xù)培養(yǎng)48 h。米托坦組：先給予米托坦干預(yù)48 h，然后將培養(yǎng)液吸出，加入少量PBS將米托坦洗去，加入不含藥物的完全培養(yǎng)液繼續(xù)培養(yǎng)48 h。聯(lián)合組根據(jù)給藥方式分為如下組別：①先給予卡非佐米后給予米托坦組(卡非佐米→米托坦組)：先給予卡非佐米干預(yù)48 h，然后將培養(yǎng)液吸出，加入少量PBS將卡非佐米洗去，再用米托坦繼續(xù)干預(yù)48 h。②卡非佐米與米托坦同期組(卡非佐米+米托坦組)：先給予卡非佐米和米托坦共同干預(yù)48 h，然后將培養(yǎng)液吸出，加入少量PBS將藥物洗去，然后加入不含藥物的完全培養(yǎng)液繼續(xù)培養(yǎng)48 h。③先給予米托坦后給予卡非佐米組(米托坦→卡非佐米組)：先給予米托坦干預(yù)48 h，然后將培養(yǎng)液吸出，加入少量PBS將米托坦洗去，再用卡非佐米干預(yù)48 h。各組藥物濃度分別設(shè)置為兩種藥物IC50值的1/8、1/4、1/2、1、2倍。參照2.3方法檢測(cè)上述各組細(xì)胞增殖抑制率。結(jié)果顯示，在IC50及2倍IC50濃度下，兩種細(xì)胞的卡非佐米→米托坦組細(xì)胞增殖抑制率均高于米托坦→卡非佐米組及卡非佐米+米托坦組，米托坦→卡非佐米組均高于卡非佐米+米托坦組，組間比較P均<0.05；其余濃度組間比較差異均無(wú)統(tǒng)計(jì)學(xué)意義(P均>0.05)。見(jiàn)表1。

表1 各組H295R、SW13細(xì)胞在IC50及2倍IC50濃度下增殖抑制率比較

注：與同種細(xì)胞同濃度米托坦+卡非佐米組比較，△P<0.05；與同種細(xì)胞同濃度米托坦組比較，#P<0.05；與同種細(xì)胞同濃度卡非佐米組比較，&P<0.05。

2.5 兩種藥物間作用的聯(lián)合指數(shù)(CI)檢測(cè) 采用CompuSyn軟件以Chou-Talaly公式[8]根據(jù)IC50及2倍IC50計(jì)算聯(lián)合組三種給藥方式下兩藥作用的聯(lián)合指數(shù)(CI)。CI值>1、=1、<1分別表示兩種藥物之間存在拮抗、疊加、協(xié)同效應(yīng)；CI值越小，表示協(xié)同效應(yīng)越大。結(jié)果顯示，在IC50及2倍IC50濃度下，兩種細(xì)胞的卡非佐米→米托坦組和米托坦+卡非佐米組CI值均<1，即存在藥效的協(xié)同作用，且卡非佐米→米托坦組兩藥的CI值均小于協(xié)米托坦+卡非佐米組(P均<0.05)；米托坦→卡非佐米組中兩藥的CI值均>1，且均高于卡非佐米→米托坦組和米托坦+卡非佐米組(P均<0.05)，即兩藥存在拮抗作用。見(jiàn)表2。

表2 H295R及SW13細(xì)胞根據(jù)IC50及2倍IC50計(jì)算所得的CI值比較

注：與卡非佐米→米托坦組比較，*P<0.05。

2.6 低濃度卡非佐米聯(lián)合米托坦對(duì)H295R及SW13細(xì)胞增殖的影響 取對(duì)數(shù)生長(zhǎng)期SW13及H295R細(xì)胞，分別以4×103個(gè)/孔接種于96孔板，隨機(jī)分為四組，對(duì)照組(僅含細(xì)胞和培養(yǎng)基)、米托坦組(25、50 μmol/L)、卡非佐米組(1 μmol/L)、卡非佐米(1 μmol/L)→米托坦(25、50 μmol/L)組。給藥方法同2.4，藥物干預(yù)48 h后按1.3方法檢測(cè)細(xì)胞增殖抑制率。結(jié)果顯示，1 μmol/L卡非佐米與25、50 μmol/L米托坦聯(lián)合的卡非佐米→米托坦組H295R及SW13細(xì)胞增殖抑制率分別高于25、50 μmol/L米托坦組(P均<0.05)。見(jiàn)表3。

表3 各組H295R、SW13細(xì)胞增殖抑制率比較

注：與同種細(xì)胞同濃度米托坦組比較，△P<0.05；與同種細(xì)胞卡非佐米組比較，▲P<0.05。

2.7 細(xì)胞周期檢測(cè) 取對(duì)數(shù)生長(zhǎng)期SW13及H295R細(xì)胞，分別以2×105個(gè)/孔接種于6孔板，置于培養(yǎng)箱內(nèi)培養(yǎng)12 h，待細(xì)胞完全貼壁生長(zhǎng)后隨機(jī)分為空白組、米托坦組、卡非佐米組、卡非佐米→米托坦組?？瞻捉M僅給予培養(yǎng)液培養(yǎng)，米托坦組、卡非佐米組、卡非佐米→米托坦組均以IC50藥物濃度干預(yù)96 h，給藥方法同2.4。采用流式細(xì)胞儀檢測(cè)細(xì)胞周期，步驟參照試劑盒說(shuō)明書(shū)。結(jié)果顯示，兩種細(xì)胞的各組G1、G2/M、S期細(xì)胞百分比比較差異均無(wú)統(tǒng)計(jì)學(xué)意義(P均>0.05)。見(jiàn)表4。

表4 各組不同周期細(xì)胞百分比比較

3 討論

泛素蛋白酶體途徑是細(xì)胞內(nèi)蛋白降解的主要途徑。與正常細(xì)胞相比，腫瘤細(xì)胞可無(wú)限增殖，其細(xì)胞周期及基因表達(dá)紊亂，促凋亡蛋白和抗凋亡蛋白表達(dá)失調(diào)，進(jìn)而抑制蛋白酶體降解途徑，細(xì)胞毒作用增強(qiáng)。蛋白酶體抑制劑可特異性地誘導(dǎo)腫瘤細(xì)胞發(fā)生毒性反應(yīng)，具有廣泛的抗腫瘤活性[6,9]。Nilubol等[10]采用定量高通量篩選技術(shù)為人ACC細(xì)胞株H295R篩選敏感藥物，發(fā)現(xiàn)該細(xì)胞對(duì)蛋白酶體抑制劑硼替佐米敏感，且硼替佐米對(duì)H295R的IC50為0.34 μmol/L，略高于臨床應(yīng)用硼替佐米體內(nèi)可達(dá)到的最高血藥濃度0.31 μmol/L。目前，關(guān)于其他類型的蛋白酶體抑制劑，特別是新一代蛋白酶體抑制劑卡非佐米對(duì)ACC影響的研究尚未見(jiàn)報(bào)道。本研究將蛋白酶體抑制劑卡非佐米體外作用于人ACC細(xì)胞株SW13及H295R，結(jié)果顯示卡非佐米可顯著抑制SW13及H295R細(xì)胞增殖，且具有時(shí)間和劑量依賴性?？ǚ亲裘讓?duì)H295R細(xì)胞和SW13細(xì)胞作用48 h的IC50均高于既往報(bào)道[11]，提示卡非佐米對(duì)SW13及H295R細(xì)胞敏感性較低，卡非佐米單藥治療ACC效果可能不佳。我們進(jìn)一步以1 μmol/L卡非佐米聯(lián)合米托坦處理ACC細(xì)胞，結(jié)果顯示，對(duì)于H295R細(xì)胞，聯(lián)合給藥組(卡非佐米 1 μmol/L→米托坦 50 μmol/L)細(xì)胞增殖抑制率可達(dá)(93.0±2.1)%，較50 μmol/L米托坦單藥組顯著提高；對(duì)于SW13細(xì)胞，聯(lián)合給藥組細(xì)胞增殖抑制率為(72.0±4.3)%，亦較米托坦單藥組提高。說(shuō)明低劑量(1 μmol/L)的卡非佐米可以作為增敏劑提高米托坦對(duì)ACC細(xì)胞的增殖抑制作用。

內(nèi)質(zhì)網(wǎng)應(yīng)激反應(yīng)是細(xì)胞的一種適應(yīng)性應(yīng)答方式。內(nèi)質(zhì)網(wǎng)應(yīng)激可產(chǎn)生兩種截然不同的結(jié)局，適當(dāng)?shù)膬?nèi)質(zhì)網(wǎng)應(yīng)激促進(jìn)細(xì)胞存活，劇烈、持續(xù)的內(nèi)質(zhì)網(wǎng)應(yīng)激則促進(jìn)細(xì)胞壞死[7,12]。米托坦是目前治療ACC最常用的藥物，內(nèi)質(zhì)網(wǎng)應(yīng)激是其抗腫瘤作用的機(jī)制之一[4]。本研究發(fā)現(xiàn)，米托坦可呈濃度依賴性抑制人ACC細(xì)胞SW13及H295R增殖，H295R細(xì)胞對(duì)米托坦較SW13細(xì)胞更敏感。作用48 h時(shí)米托坦對(duì)H295R和SW13細(xì)胞的IC50(分別為18.42 μmol/L和62.37 μmol/L)均在臨床推薦的血藥濃度范圍(43.75～87.5 μmol/L)內(nèi)[13]，與其他研究結(jié)果相似[14]?？ǚ亲裘缀兔淄刑箍鼓[瘤作用的機(jī)制均包括內(nèi)質(zhì)網(wǎng)應(yīng)激，因此推測(cè)二者聯(lián)用可能增加米托坦的抗腫瘤作用。本研究中卡非佐米和米托坦聯(lián)合應(yīng)用對(duì)H295R和SW13細(xì)胞存在序貫依賴性的協(xié)同作用。以卡非佐米+米托坦、卡非佐米→米托坦及米托坦→卡非佐米三種給藥方式處理H295R和SW13細(xì)胞48 h，MTT檢測(cè)結(jié)果顯示，在IC50和2倍IC50濃度條件下，卡非佐米+米托坦及卡非佐米→米托坦兩種給藥方式均可明顯抑制H295R和SW13細(xì)胞增殖，但后者的抑制作用更顯著；兩種給藥方式下兩藥的CI值均<1，提示存在藥效的協(xié)同作用。

目前關(guān)于抗腫瘤藥物序貫依賴性的抗增殖作用機(jī)制尚不完全清楚，干擾細(xì)胞周期分布可能與其有關(guān)。兩種藥物序貫給藥時(shí)，若前一種將細(xì)胞阻滯在G1期，則將影響接下來(lái)細(xì)胞周期特異性毒性藥物的作用[15]。有文獻(xiàn)報(bào)道，卡非佐米可以將甲狀腺癌細(xì)胞周期阻滯在G2/M期，與其他抗腫瘤藥物聯(lián)合應(yīng)用時(shí)不同給藥方案可能產(chǎn)生不同的效果[16]。本研究發(fā)現(xiàn)，給予IC50濃度的卡非佐米和米托坦單藥或序貫聯(lián)合處理SW13、H295R細(xì)胞，G1、G2/M、S期細(xì)胞百分比比較差異均無(wú)統(tǒng)計(jì)學(xué)意義。說(shuō)明卡非佐米和米托坦序貫用藥對(duì)細(xì)胞增殖抑制作用的差異可能并不是通過(guò)影響細(xì)胞周期來(lái)實(shí)現(xiàn)的。Kroiss等[17]研究發(fā)現(xiàn)，卡非佐米可以通過(guò)增加IRE1磷酸化，引起其下游通路XBP1裂解。XBPI是一種轉(zhuǎn)錄因子，其可進(jìn)入細(xì)胞核啟動(dòng)應(yīng)激反應(yīng)基因如CHOP、GRP78等轉(zhuǎn)錄，進(jìn)而完成內(nèi)質(zhì)網(wǎng)應(yīng)激反應(yīng)。米托坦則可以同時(shí)增加eIF2α和IRE1磷酸化，導(dǎo)致促凋亡蛋白如Noxa、CHOP、DR5等表達(dá)增加，引起細(xì)胞凋亡。但本研究卡非佐米聯(lián)合50 μmol/L米托坦處理H295R細(xì)胞時(shí)，內(nèi)質(zhì)網(wǎng)應(yīng)激蛋白CHOP蛋白表達(dá)和XBP1裂解均較聯(lián)合25 μmol/L米托坦時(shí)減少，這可能是不同的序貫用藥順序?qū)?xì)胞增殖抑制作用差異的主要原因。

綜上所述，卡非佐米聯(lián)合米托坦對(duì)ACC細(xì)胞存在序貫依賴性的協(xié)同抑制細(xì)胞增殖的作用，先給予卡非佐米后給予米托坦的給藥方式的抗增殖作用最佳，但兩藥聯(lián)合對(duì)細(xì)胞周期無(wú)明顯影響。

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EffectsofcarfilzomibcombinedwithmitotaneonproliferationandcellcycleofhumanadrenocorticalcarcinomaH295RandSW13cells

WEIQingchen,TANRuyin,YANGHaiyan,HUANGZhenxing,LIANGXinghuan,QINYingfen,LUOZuojie

(TheFirstAffiliatedHospitalofGuangxiMedicalUniversity,Nanning530021,China)

ObjectiveTo investigate the effects of carfilzomib combined with mitotane on the proliferation and cell cycle of human adrenocortical carcinoma (ACC) H295R and SW13 cells.MethodsHuman ACC H295R and SW13 cells were cultured in vitro. Using MTT to determine the IC50values of mitotane on H295R and SW13 cells at 48 h, which were 18.42 and 62.37 μmol/L, respectively, and those of carfilzomib were 3.86 and 11.62 μmol/L. These two cell lines in the logarithmic growth phase were randomly divided into the control group, carfilzomib group, mitotane group, and the carfilzomib → mitotane group, carfilzomib + mitotane group, and the mitotane →carfilzomib group (namely according to the sequential administration). The concentrations of the two drugs were 1/8, 1/4, 1/2, 1, 2 times of IC50values, respectively. The inhibitory rate of cell proliferation was measured at 48 h. The combine index (CI) of the two drugs was calculated according to the Chou-Talalay formula. These two cells in the logarithmic growth phase were randomly divided into the control group, mitotane monotherapy group (25 and 50 μmol/L), carfilzomib monotherapy group (1 μmol/L), carfilzomib (1 μmol/L) → mitotane (25 and 50 μmol/L) group. The inhibitory rate of cell proliferation at 48 h after drug intervention was detected. These two cells in the logarithmic growth phase were randomly divided into the blank group, mitotane group, carfilzomib group, carfilzomib → mitotae group, and the latter three groups were intervened at the drug concentration of IC50for 96 h, and flow cytometry was used to detect the cell cycle.ResultsThe proliferation inhibitory rates of H295R and SW13 cells were significantly higher in the carfilzomib→mitotane group than in the mitotane→carfilzomib group and the carfilzomib+ mitotane group at the concentrations of IC50and 2-fold IC50; the mitotane→carfilzomib group was higher than the carfilzomib+ mitotane group; there were significant differences between every two groups (allP<0.05). CI value<1 was found in both the carfilzomib→mitotane group and the mitotane + carfilzomib group at the concentrations of IC50and 2-fold IC50in H295R and SW13 cell lines, and the CI value in the carfilzomib→mitotane group was lower than that in the mitotane + carfilzomib group (allP<0.05); CI value >1 was found in the mitotane→carfilzomib group, which was greater than that of the carfilzomib→mitotane group and the mitotane + carfilzomib group (allP<0.05). The proliferation inhibitory rates of H295R and SW13 cells were significantly higher in the carfilzomib (1 μmol/L)→mitotane (25 and 50 μmol/L) group than in the mitotane group (25 and 50 μmol/L), respectively. There was no significant difference in the percentage of cells in the G1, G2/M, and S phases between these four groups (P>0.05).ConclusionThere are sequence-dependent antiproliferative effects of mitotane and carfilzomib on H295R and SW13 cell lines, and the sequential administration of carfilzomib followed by mitotane exhibits the strongest anticancer activity, but the combination of these two drugs has no significant effect on the cell cycle.

adrenocortical carcinoma; carfilzomib; mitotane; cell proliferation; cell cycle

國(guó)家自然科學(xué)基金資助項(xiàng)目(81060220)。

韋慶臣(1990-)，男，住院醫(yī)師，研究方向?yàn)閮?nèi)分泌代謝疾病及相關(guān)腫瘤。E-mail: 947343812@qq.com

羅佐杰(1958-)，男，教授，研究方向?yàn)閮?nèi)分泌代謝疾病及相關(guān)腫瘤。E-mail: zluo888@163.com

10.3969/j.issn.1002-266X.2017.48.007

R736.6

A

1002-266X(2017)48-0023-05

2017-05-14)