【摘要】：目的研究亞致死溫度射頻消融(RFA)對(duì)肝癌干細(xì)胞(LCSC)產(chǎn)生及其相關(guān)轉(zhuǎn)錄因子表達(dá)的影響。方法利用小鼠Hep1-6肝癌細(xì)胞株和肝細(xì)胞癌(HCC)患者臨床樣品,檢測(cè)LCSC相關(guān)標(biāo)志物和轉(zhuǎn)錄因子的表達(dá)。結(jié)果不同溫度分別刺激Hep1-6細(xì)胞后發(fā)現(xiàn),45℃是不能誘導(dǎo)細(xì)胞死亡的亞致死性溫度。流式細(xì)胞儀(FCM)檢測(cè)顯示45℃處理可引起CD13~+、CD44~+、CD90~+、CD133~+Hep1-6細(xì)胞水平明顯上調(diào),提示45℃溫度導(dǎo)致Hep1-6細(xì)胞中以上各型LCSC產(chǎn)生增加;實(shí)時(shí)定量聚合酶鏈反應(yīng)(RT-q PCR)檢測(cè)顯示45℃溫度導(dǎo)致CD13、CD90、CD133 m RNA水平明顯上調(diào)。CD13 m RNA水平在5例HCC患者復(fù)發(fā)肝癌組織中均明顯上調(diào),CD133 m RNA在4例復(fù)發(fā)肝癌中上調(diào),CD90 m RNA僅在1例復(fù)發(fā)肝癌中上調(diào);FCM檢測(cè)顯示CD13~+LCSC水平在4例復(fù)發(fā)肝癌中明顯上調(diào),CD133~+LCSC水平僅在1例復(fù)發(fā)肝癌中上調(diào),提示CD13~+LCSC水平上調(diào)與45℃溫度關(guān)系更密切。RT-q PCR檢測(cè)顯示4例CD13~+LCSC上調(diào)的復(fù)發(fā)肝癌患者13個(gè)LCSC相關(guān)轉(zhuǎn)錄因子中Sox2、Stat1明顯上調(diào),FCM檢測(cè)顯示45℃處理Hep1-6細(xì)胞后Sox2、Stat1 m RNA明顯上調(diào)。用Sox2、Stat1 si RNA分別沉默了Sox2、Stat1基因,表明Sox2、Stat1均參與了45℃溫度誘導(dǎo)的CD13~+LCSC產(chǎn)生。結(jié)論 RFA治療中45℃亞致死性溫度所致CD13~+LCSC水平增高與Sox2、Stat1表達(dá)有關(guān)。該結(jié)果對(duì)肝癌復(fù)發(fā)研究有一定借鑒意義。
[Abstract]:Objective to study the effect of sublethal temperature radiofrequency ablation (RFA) on (LCSC) production and transcription factor expression in hepatocellular carcinoma stem cells. Methods the expression of LCSC related markers and transcription factors were detected by using mouse Hep1-6 hepatoma cell line and clinical samples of (HCC) patients with hepatocellular carcinoma. Results after stimulation of Hep1-6 cells at different temperatures, it was found that 45 鈩，

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