Mx1基因表达对VSV溶瘤敏感性的影响  

Effects of Mxl gene expression on oncolytic sensitivity of murine tumor cells towards VSV

作　　者：刘林 丁立伟 严专 郭秉楠 

机构地区：[1]徐州医科大学急救与救援医学系,江苏徐州221002

出　　处：《徐州医学院学报》2016年第12期779-783,共5页Acta Academiae Medicinae Xuzhou

基　　金：国家自然科学基金（81471586）;徐州市科技计划项目（XM128025,KC14SH087）

摘　　要：目的研究3种小鼠肿瘤细胞系鼠黏病毒蛋白1（Mx1）基因表达对水泡性口膜炎病毒（vesicular stomatitis virus，VSV）溶瘤效应的敏感性。方法将本实验室定量后的VSV保种并用来攻击3种不同种类的肿瘤细胞系：RAW264．7（小鼠巨噬细胞白血病）、B16（黑色素瘤）、HEPA1—6（小鼠肝癌）细胞系，48h后通过TCID，。法和MTI＂法检测3种肿瘤细胞系对VSV的敏感性。RT—PCR检测VSV感染3种肿瘤细胞系24h后Mx1基因的诱导表达水平，证实肿瘤细胞系的溶瘤敏感性与Mx1基因表达的关系；应用RNAi技术抑制Mx1基因的表达水平，MTT法检测RAW264．7细胞系（最不敏感细胞系）与B16细胞系（最敏感细胞系）溶瘤效果的改变。结果用VSV攻击3种肿瘤细胞系呈现出的敏感性依次为B16〉HEPA1—6〉RAW264．7。通过RT—PCR检测VSV感染3种鼠源肿瘤细胞系Mx1基因的表达水平表明：Mx1基因的表达水平越高，VSV对肿瘤细胞的溶瘤敏感性越低。应用RNAi技术抑制RAW264．7细胞系（最不敏感细胞系）Mx1基因的表达，VSV对该细胞系溶瘤敏感性提高了35％，抑制B16细胞系（最敏感细胞系）Mx1基因的表达，VSV对该细胞系溶瘤的敏感性提高了30％。结论通过RNAi抑制鼠源肿瘤细胞系抗病毒蛋白Mx1基因的表达水平，可促进VSV溶瘤的敏感性。Objective To investigate the effects of mouse myxovirus （Mxl） expression on the sensitivities of three mouse tumor cell lines to oncolytic vesicular stomatitis virus （VSV）. Methods The VSV strains preserved in our labo- ratory were adopted to challenge a mouse macrophage - like RAW264.7 cell line, a melanoma B16 cell line, and a mouse hepatoma Hepal - 6 cellline. Then, 48 hours later, the three types of cells were tested for their sensitivity to VSV by TCIDs0 and MTI＂ assay. The induced levels of Mxl gene in the three cell lines infected with VSV for 24 hours were de- tected by RT - PCR. Furthermore, the changes in the oncolytic effects of RAW264.7 and B16 cells with inhibited Mxl gene expression by RNAi were determined by MTY assay. Results The sequence of the sensitivity towards VSV chal- lenge was shown as follows： B16 〉 HEPA1 -6 〉 RAW264.7. According to RT -PCR results, the higher the Mxl gene level was, the lower sensitivity of the tumor cells towards VSV. The expression of Mxl gene in RAW264.7 cells was in- hibited by RNAi techniques, where the sensitivity of RAW264.7 cells towards VSV was increased by 35%. The expres- sion of Mxl gene in B16 cells was inhibited by RNAi techniques, where the sensitivity of B16 cells towards VSV was in- creased by 30%. Conclusions Suppression of antiviral gene Mxl expression by RNAi can improve the serisitivity of the tumor cells to VSV.

关 键 词：鼠黏病毒蛋白1 水泡性口膜炎病毒 溶瘤 RNA干扰 

分 类 号：R738.7[医药卫生—肿瘤]