反义局部粘着斑激酶抑制肝癌侵袭生长的研究

Effects of antisense FAK ODN transfection on xenograft hepatocellular carcinoma in nude mice

目的 观察反义局部粘着斑激酶 (FAK)脱氧寡核苷酸 (ODN)转染对Bel740 2肝癌移植瘤侵袭性生长的影响 ,并探讨其作用机制。方法 以LipofectAMINE介导的反义FAKODN转染Bel 740 2肝癌细胞株后 ,于 6只裸鼠双侧颈背部、髋部共 4处皮下接种 ,观察移植瘤生长情况 ,并行肿瘤微血管密度 (MVD)的免疫组织化学检测及MMP2与TIMP2表达的逆转录 聚合酶链反应(RT PCR)检测。结果 反义FAKODN转染使裸鼠皮下移植瘤的生长受到显著抑制 ,抑瘤率为3 5 .7% ;MVD在反义转染组 ( 9.5 99± 1.2 84)显著低于对照组 [( 13 .915± 2 .618) ,P <0 .0 5 ] ;MMP2表达在反义转染组 ( 0 .199± 0 .0 61)显著低于对照组 ( 0 .42 1± 0 .118) ,TIMP2表达在反义转染组( 0 .461± 0 .15 3 )则显著高于对照组 [( 0 .2 98± 0 .10 4) ,P <0 .0 5 ]。结论 信号转导分子FAK表达阻断可显著抑制Bel 740 2肝癌细胞株在裸鼠体内的侵袭性生长 ,病理性肿瘤血管生成减少及MMP2、TIMP2表达改变与其抗肿瘤作用密切相关。

Objective To study the in vivo effects of antisense FAK ODN transfection on the tumorigenicity of Bel 7402 hepatocellular carcinoma cells,and to investigate the underlying mechanism.Methods LipofecTAMINE-mediated antisense FAK oligodeoxynucleotides (ODN) were transfected into Bel 7402 cells.These cells were then inoculated subcutaneously in 4 areas on nape and hip of 6 nude mice.The growth and invasion of the xenograft hepatocellular carcinoma were observed.The transplant tumor was obtained after experiment and MCV with immunohistochemical staining and MMP2,TIMP2 with RT-PCR were detected.Results Tumor growth in antisense FAK ODN treated nude mice was significantly retarded in comparison with control mice.The weight of the tumor was markedly lighter in transfected mice (126.0±12.6) mg than that in control mice (196.0±28.1) mg(P<0.05),with the inhibitory rate of tumor growth beomg (35.71±14.20)%;The expression levels of MCV and MMP2 in transplanted group (9.599±1.284,0.199±0.061,respectively) were significantly lower than those in control group (13.915±2.618,0.421±0.118,respectively),while TIMP2 expression level in transplanted group (0.461±0.153) was significantly higher than that in control group [(0.298±0.104),P<0.05].Conclusion Suppression of the signal transduction molecule-FAK expression can significantly inhibit the tumor growth of Bel 7402 cells in vivo.Decreased pathological angiogenesis and reduced ratio of MMP2/TIMP2 play an important part in this process.