AAV-HGFK1抑制EGFR磷酸化拮抗大肠癌细胞生长  

AAV-HGFK1 inhibits proliferation of colorectal cancer cells by down-regulating the phosphorylation of EGFR

作　　者：邓飞鸿 聂飚[1] 左俊华[1] 柳雪花 陈金敏[1] 

机构地区：[1]南方医科大学南方医院消化科,广州510515

出　　处：《重庆医学》2014年第33期4447-4449,4453,共4页Chongqing medicine

摘　　要：目的研究腺相关病毒-肝细胞生长因子K1(AAV-HGFK1)对KRAS、BRAF野生型和突变型大肠癌细胞株的生长影响。方法选择人源的细胞株KRAS和BRAF野生的SW48,KRAS突变的Lovo,KRAS突变且不表达表皮生长因子受体(EGFR)的SW620,BRAF突变的HT29,实时荧光定量PCR(RT-PCR)测定EGFR基因转录水平,分别用5×104 vg/细胞的腺相关病毒(AAV)感染以上细胞株,荧光显微镜和流式细胞仪测定AAV-EGFP感染率。加入或不加表皮生长因子(EGF)的情况下,蛋白免疫印迹法(Western blot)检测EGFR、磷酸化EGFR(p-EGFR)和β-actin,四甲基偶氮唑盐比色(MTT)法测细胞增殖。结果Lovo和HT29高表达EGFR,而SW48表达EGFR较低,加入EGF促进了以上3种细胞的EGFR磷酸化,AAV-HGFK1抑制EGFR磷酸化,从而显著降低其增殖。EGF对SW620增殖无显著影响,但AAV-HGFK1感染抑制了胎牛血清培养的SW620细胞。结论 AAV-HGFK1可能直接作用于EGFR,抑制其磷酸化从而阻断EGF促进细胞增殖,并可能通过其他通路对KRAS或BRAF基因野生或突变的大肠癌细胞都有抑制作用。Objective To study the effect of adeno associated virus hepatocyte growth factor K1（AAV‐HGFK1）on the prolifer‐ation of 4 different colorectal cell lines with or without KRAS or BRAF mutation .Methods The levels of epidermal growth factor receptor （EGFR） mRNA were determined in SW48 without KRAS or BRAF mutation ,Lovo with KRAS mutation ,SW620 with KRAS mutation ,HT29 with BRAF mutation by quantitative real time PCR ,respectively .After the infection of AAV‐HGFK1 ,the expressions of EGFR ,p‐EGFR and β‐actin were detected by Western blot and the proliferation of the cells were assayed using MTT .Results Lovo ,SW48 and HT29 expressed EGFR protein while SW620 did not .EGF promoted the proliferation of Lovo , SW48 and HT29 cells .AAV‐HGFK1 down‐regulated the phosphorylation of EGFR and significantly inhibited their proliferation . But EGF had no effect on proliferation of SW620 stimulated by EGF .Conclusion AAV‐HGFK1 exhibited its antitumor effects through EGF/EGFR signaling irrespective of the KRAS or BRAF mutation and may also act through other signaling pathways .

关 键 词：受体 表皮生长因子 大肠肿瘤 肝细胞生长因子K1 腺相关病毒 

分 类 号：R735.34[医药卫生—肿瘤]