摘要
目的:观察转染ITAC对乳腺癌细胞系4T1体内及体外生物学行为的影响。方法:构建pcDNA3-ITAC真核表达质粒,基因转染的方法建立稳定表达ITAC的乳腺癌细胞系ITAC-4T1。体外及体内观察ITAC-4T1细胞的生长情况。RT-PCR检测肿瘤组织ITACmRNA转录水平。杀伤实验检测脾细胞杀伤活性,FACS检测CD8+T细胞IFN-γ的分泌。结果:体外ITAC-4T1细胞的生长与未转染及转染空载体的4T1细胞没有差别(P>0.05)。体内ITAC-4T1细胞形成的肿瘤生长较对照组明显减慢(P<0.05)。ITAC-4T1肿瘤组织检测到较高水平的ITACmRNA转录(P<0.01)。接种ITAC-4T1细胞的小鼠脾细胞杀伤率显著高于对照组(P<0.05),CD8+T细胞IFN-γ分泌明显增加(P<0.05)。结论:ITAC基因转染可以抑制4T1细胞体内生长,与ITAC诱导Th1型细胞免疫应答,增强效应细胞特异性杀伤活性相关。
Purpose:To investigate the cellular characteristics of ITAC(IFN-γ inducible T cell α chemoattractant) transfected 4T1 mammary carcinoma cell line in vivo and in vitro. Methods:pcDNA3-ITAC plasmid was constructed. A mammary carcinoma cell line 4T1 was transfected with pcDNA3-ITAC plasmid and the positive clones (ITAC-4T1) were screened with G418. Growth kinetics of ITAC-4T1 cells was observed in vitro and in vivo. ITAC mRNA transcription in tumor tissues was examined by RT-PCR. Cytotoxicity of splenocytes was analyzed by cytotoxic assay. Expression of IFN-γ by CD8~ + T cells was tested by intracellular cytokine staining. Results:The growth rate of ITAC-4T1 cells was similar to that of the parental 4T1 cells and neo-vector transfected 4T1 cells in vitro. Growth of the tumors formed by ITAC-4T1 cells was dramatically inhibited in vivo (P<0.05). High expression of ITAC mRNA was detectable in ITAC-4T1 tumor tissues. After being stimulated with the specific antigen, splenocytes from mice inoculated with ITAC-4T1 cells showed vigorous cytotoxic activity compared to those from controls (P<0.05). The expression of IFN-γ was increased significantly in CD8~ + T cells from mice inoculated with ITAC-4T1 cells (P<0.05). Conclusions:Expression of ITAC in tumors might exert a local anti-tumor function by enhancing Th1-type cellular immune response leading to cytotoxicity against specific tumor cells.
出处
《中国癌症杂志》
CAS
CSCD
2005年第3期252-256,共5页
China Oncology
基金
国家教育部科学技术重点项目(03063)
国家自然科学基金(3017086)。
