【摘要】：目的:研究結(jié)腸癌轉(zhuǎn)移相關(guān)基因1(metastasis associated in colon cancer1,MACC1)對(duì)人鼻咽癌細(xì)胞惡性生物學(xué)行為的影響,觀察人鼻咽癌細(xì)胞過(guò)表達(dá)MACC1對(duì)細(xì)胞的增殖、遷移及侵襲的影響。方法:1.采用Western blot技術(shù)檢測(cè)鼻咽癌5-8F、HNE-1、CNE-1、CNE-2細(xì)胞株中MACC1的蛋白表達(dá)水平。2.通過(guò)慢病毒轉(zhuǎn)染技術(shù)建立一組過(guò)表達(dá)MACC1的CNE-2細(xì)胞株,篩選出穩(wěn)定轉(zhuǎn)染的細(xì)胞。3.實(shí)驗(yàn)分為三組:實(shí)驗(yàn)組(CNE-2 MACC1過(guò)表達(dá)穩(wěn)定株組);陰性對(duì)照組(CNE-2轉(zhuǎn)染空載體穩(wěn)定株組);空白對(duì)照組(CNE-2空細(xì)胞組)。4.采用Q-PCR實(shí)驗(yàn)和Western blot實(shí)驗(yàn)檢測(cè)三組細(xì)胞MACC1的表達(dá)水平。5.采用MTT實(shí)驗(yàn)觀察各組細(xì)胞的增殖能力。6.采用Transwell遷移實(shí)驗(yàn)觀察各組細(xì)胞的遷移能力。7.采用Transwell侵襲實(shí)驗(yàn)觀察各組細(xì)胞的侵襲能力。結(jié)果:1.Western blot實(shí)驗(yàn)結(jié)果顯示HNE-1細(xì)胞株表達(dá)MACC1的水平最高,CNE-1細(xì)胞株表達(dá)水平中等,CNE-2和5-8F細(xì)胞株未檢測(cè)到MACC1的表達(dá)或表達(dá)量極低。2.慢病毒轉(zhuǎn)染后,在倒置熒光顯微鏡下觀察到實(shí)驗(yàn)組細(xì)胞有綠色熒光表達(dá)。3.Q-PCR實(shí)驗(yàn)結(jié)果顯示,實(shí)驗(yàn)組細(xì)胞的MACC1 mRNA的表達(dá)量遠(yuǎn)高于對(duì)照組。4.Western blot結(jié)果顯示,實(shí)驗(yàn)組細(xì)胞條帶明顯,有MACC1蛋白的過(guò)表達(dá),而對(duì)照組細(xì)胞未見(jiàn)條帶,提示MACC1無(wú)表達(dá)或表達(dá)量極低。5.MTT結(jié)果顯示,實(shí)驗(yàn)組細(xì)胞在慢病毒轉(zhuǎn)染后24h、48h、72h的增殖能力與對(duì)照組相比明顯增強(qiáng),差異有統(tǒng)計(jì)學(xué)意義(P0.05)。6.Transwell遷移實(shí)驗(yàn)結(jié)果顯示,培養(yǎng)24h后,實(shí)驗(yàn)組細(xì)胞的遷移數(shù)為(85±4)個(gè),陰性對(duì)照組細(xì)胞的遷移數(shù)為(59±3)個(gè),空白對(duì)照組細(xì)胞的遷移數(shù)為(58±4)個(gè),實(shí)驗(yàn)組與對(duì)照組之間差異有統(tǒng)計(jì)學(xué)意義(P0.05)。7.Transwell侵襲實(shí)驗(yàn)結(jié)果顯示,培養(yǎng)24h后,實(shí)驗(yàn)組細(xì)胞的侵襲數(shù)為(81±4)個(gè),陰性對(duì)照組為(42±3)個(gè),空白對(duì)照組為(40±4)個(gè),實(shí)驗(yàn)組與對(duì)照組之間差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:MACC1的異常高表達(dá)會(huì)導(dǎo)致鼻咽癌CNE-2細(xì)胞的增殖、遷移和侵襲等惡性生物學(xué)行為增強(qiáng),提示MACC1參與了鼻咽癌的X椫場(chǎng)⑶窒�、转翌l戎琢齜⑸⒄溝墓獺

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