敲低AKT1基因增强裸鼠胃癌移植瘤对阿霉素的敏感性

发布时间：2018-06-20 12:46

本文选题：AKT + 慢病毒载体　；参考：《细胞与分子免疫学杂志》2017年10期

【摘要】：目的研究敲低AKT1基因对裸鼠胃癌移植瘤阿霉素敏感性的影响。方法构建AKT1基因RNA干扰(RNAi)的慢病毒载体p GCSIL-sh AKT1-GFP,转染HEK293T细胞,以p GCSIL-sh CON-GFP为空载体组。获得慢病毒颗粒后,感染胃癌BGC-823细胞,采用Western blot法检测感染后BGC-823细胞中AKT1蛋白水平。皮下注射慢病毒感染的BGC-823细胞构建裸鼠胃癌移植瘤模型,使用阿霉素(DOX)进行处理,观察处理后移植瘤体积大小并绘制肿瘤生长曲线,采用HE染色观察移植瘤组织病变情况,采用组织原位末端转移酶标记技术(TUNEL)检测移植瘤组织中细胞凋亡情况。结果成功构建AKT1基因RNAi的慢病毒载体并成功感染胃癌BGC-823细胞,感染后胃癌BGC-823细胞中AKT1蛋白水平明显降低。敲低AKT1的BGC-823细胞裸鼠移植后,移植瘤生长减慢、移植瘤细胞凋亡增加,并可增强DOX对移植瘤生长的抑制作用。结论敲低AKT1基因可增强BGC-823细胞裸鼠移植瘤对DOX的敏感性。
[Abstract]:Objective to study the effect of knock-down AKT1 gene on adriamycin sensitivity in nude mice with gastric cancer xenografts. Methods the lentivirus vector pGCSIL-sh AKT1-GFP1 was constructed and transfected into HEK293T cells. The vector pGCSIL-sh CON-GFP was used as empty vector. BGC-823 cells were infected with lentivirus particles, and the AKT1 protein levels in BGC-823 cells were detected by Western blot assay. BGC-823 cells infected with lentivirus were injected subcutaneously into nude mice gastric cancer transplanted tumor model, treated with doxorubicin (DOX), the volume of transplanted tumor was observed and the tumor growth curve was plotted, and the pathological changes of transplanted tumor were observed by HE staining. Tunel was used to detect apoptosis in transplanted tumor tissue. Results the lentivirus vector of AKT1 gene RNAi was successfully constructed and successfully infected with gastric cancer BGC-823 cells. The level of AKT1 protein in BGC-823 cells decreased significantly after infection. After transplanting BGC-823 cells with low AKT1, the growth of xenografts slowed down, the apoptosis of transplanted tumor cells increased, and the inhibitory effect of DOX on the growth of xenografts was enhanced. Conclusion knock down AKT1 gene can enhance the sensitivity of BGC-823 xenografts to DOX.
【作者单位】：武汉大学人民医院消化内科;
【基金】：国家自然科学基金(81401959)
【分类号】：R735.2

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