TIM2基因修饰的H22细胞体内成瘤作用的研究

Study on oncogenicity of H22 cells modified by TIM2 gene in vivo

目的观察小鼠TIM2基因修饰的H22肝癌细胞在小鼠体内的成瘤作用,初步研究TIM2基因在肿瘤生物治疗中的作用。方法构建TIM2基因的真核表达载体pIRES2-EGFP-TIM2,脂质体法转染H22细胞,体外筛选得到稳定表达TIM2基因的阳性单克隆H22细胞株,用以建立小鼠肝癌移植瘤模型,同时以转染了pIRES2-EGFP空载体的H22-EGFP细胞和H22细胞做为对照,观察不同处理的H22细胞对小鼠的成瘤情况。结果重组真核表达载体pIRES2-EGFP-TIM2转染H22细胞后,经体外筛选和鉴定,得到稳定表达EGFP和TIM2基因的阳性单克隆细胞株H22-TIM2,免疫接种小鼠后可明显抑制小鼠肿瘤的发生和发展,小鼠肿瘤组织块的体积明显小于接种H22-EGFP细胞和H22细胞组。此外,接种H22-TIM2细胞组小鼠的生长情况也明显好于其他各组。结论TIM2基因转染H22细胞后可显著降低小鼠H22肝癌细胞的成瘤性,抑制小鼠体内肿瘤的生长。本研究为进一步探讨TIM2在肿瘤生物治疗中的作用提供了初步的实验依据。

[Objective] To investigate the effect of routine hepatocarcinoma cells H22 modified by TIM2 gene on tumor formation in mice, and explore the usage of TIM2 gene in the biotherapy of tumors in vivo. [Methods] The combinant eukaryotic expression vector pIRES2-EGFP-TIM2 was constructed and transfected into H22 ceils by lipofectamin. The H22-TIM2 ceils with TIM2 gene expression was obtained by stable electing essay in vitro and the expressed EGFP protein was observed under fluorescent microscope and the TIM2 mRNA expression was detected by RT-PCR. Then the H22-TIM2 cells were used to establish the tumor-bearing mice model. The H22-EGFP ceils transfected with pIRES2-EGFP vector and H22 cells were also injected subcutaneously into mice as the experimental control. The oncogenicity of H22 cells modified by TIM2 gene was observed and the effect on tumor growth was compared with H22-EGFP cells and H22 cells in mice. [Results] The co-expression of EGFP and TIM2 gene was detected in TIM2 gene transfered in H22 cells, Transfected by pIRES2-EGFP-TIM2 vector, H22-TIM2 cells could coexpressethe EGFP and TIM2 gene. The formation and growth of tumor were significantly inhibited in the group injected with H22-TIM2 cells. The volumes of tumor in H22-TIM2 group mice were smaller than those two groups of mice with H22-EGFP cells and H22 cells, respectively. Morever, the general status and activity in H22-TIM2 group were obviously better than the other two groups. [Conclusions] The tumorigenesis of H22 cells is markedly impaired after being transfected of TIM2 gene. Modification of H22 cells with TIM2 gene can suppress the growth of tumor in mice. Our results provide a preliminary study for biotherapy of tumor with TIM2 gene.