重组溶瘤单纯疱疹病毒对小鼠结肠癌移植瘤的抑制作用

Inhibitory Effect of Recombinant Oncolytic Herpes Simplex Virus on the Transplanted Tumor of Colon Cancer in Mice

探讨表达p53和IL-12基因的重组溶瘤单纯疱疹病毒(oncolytic Herpes Simplex Virus,oHSV)MH1004和MH-1006对小鼠结肠癌移植瘤的抑制作用.分别以Western blot和ELISA检测MH1004和MH1006感染细胞中治疗基因p53和IL-12的表达.以结肠癌细胞CT26构建小鼠皮下移植瘤模型,瘤内注射HSV-wt、HSV空载体MH1001和MH1005以及oHSV MH1004和MH1006,观察oHSV对小鼠肿瘤生长、存活率及肿瘤大小的影响,结果显示:Western blot可检测到MH1004感染细胞中p53蛋白的表达;ELISA检测表明MH1006在感染细胞早期表达IL-12蛋白.荷瘤小鼠治疗第21d,MH1004、MH1005和MH1006治疗组肿瘤体积依次为2820.69±2539.35 mm^(3)、2127.31±2017.02 mm^(3)和1414.36±1639.19 mm^(3),均显著小于Mock组(7682.92±2648.74 mm^(3))(P<0.01).治疗42 d时,MH1004、MH1005和MH1006组均有1只小鼠肿瘤完全消失.小鼠生存率结果表明:MH1004(100%)和MH1006组(100%)显著高于MH1001(66.67%)、MH1005(66.67%)、HSV-wt(50%)和Mock组(50%)(P<0.05).以上研究说明MH1005是一种理想的复制型溶瘤病毒载体;表达p53和IL-12的oHSV能够有效抑制小鼠体内结肠癌肿瘤的生长,是一种潜在的结肠癌治疗新方法.

To study the inhibitory effect of the recombinant oncolytic herpes simplex virus(oHSV)MH1004 and MH1006 expressing p53 and IL-12 genes on the transplanted colon cancer tumors in mice.Western blot and ELISA were used to detect the expression of therapeutic gene p53 and IL-12 in the cells infected with MH1004 and MH1006 respectively;Colon cancer cell CT26 was used to construct the subcutaneous transplanted tumor model in mice.HSV-wt,HSV empty viral vector MH1001 and MH1005,as well as oHSV MH1004 and MH1006 were injected into the tumor to observe the effect of the oHSVs on tumor growth,survival rate and tumor size in the mice.The results were shown as below,p53 protein was detected by Western blot in the cells infected with MH1004,and IL-12 protein was detected by ELISA in the cells infected with MH1006 in the early stage.At day 21 of treatment for tumor bearing mice,the tumor volumes of MH1004,MH1005 and MH1006 treatment groups were 2820.69±2539.35 mm^(3),2127.31±2017.02 mm^(3) and 1414.36±1639.19 mm^(3) respectively,which were all significantly smaller than that of Mock group(7682.92±2648.74 mm^(3))(P<0.01).At day 42 of the treatment,the tumor in one of the mice from MH1004,MH1005 and MH1006 groups all disappeared respectively,and the survival rates of mice of MH1004(100%)and MH1006(100%)groups were significantly higher than that of MH1001(66.67%),MH1005(66.67%),HSV-wt(50%)and Mock(50%)groups(P<0.05).This study suggests MH1005 is an ideal replication oncolytic viral vector;oHSV expressing p53 and IL-12 can effectively inhibit the growth of colon cancer tumors in mice,which is a potential new method for colon cancer treatment.