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目的:利用反义核酸技术,研究体外胰岛素样生长因子-Ⅰ(IGF-Ⅰ)反义寡核苷酸转染对肝癌细胞增生、分化及凋亡的影响,探讨寡核苷酸转染治疗肝癌的可行性. 方法:合成针对IGF-Ⅰ的寡核苷酸片段,利用脂质体包裹反义IGF-Ⅰ寡核苷酸片段瞬时转染人肝癌细胞系BEL-7402细胞,MTT法检测细胞增生;放免法检测培养细胞上清中AFP,CEA的分泌量;免疫组化法检测转染细胞AFP, PCNA表达的变化;采用末端标记(Tunel)法检测细胞凋亡的变化. 结果:MTT法检测转染反义IGF-Ⅰ寡核苷酸的人肝癌细胞系BEL-7402 A值由0.44±0.09下降为0.30±0.07(P<0.01), 上清液AFP和CEA水平分别由12.5±2.2μg/L和6.8±2.3μg/L下降为2.5±0.3μg/L和2.2±1.5μg/L(P<0.01, P<0.05),凋亡阳性细胞数由16.4±2.3%增加至23.1±3.7%(P<0.05). 结论:反义IGF-Ⅰ寡核苷酸转染人肝癌细胞系BEL-7402细胞可以降低细胞增生,减少细胞去分化并诱导肝癌细胞凋亡.
OBJECTIVE: To study the effect of antisense oligonucleotide transfection with insulin-like growth factor-I (IGF-Ⅰ) antisense oligonucleotide on the proliferation, differentiation and apoptosis of hepatocellular carcinoma cells in vitro and to explore the effect of oligonucleotide transfection on hepatoma .Methods: The oligonucleotide fragments targeting IGF-Ⅰ were synthesized and transfected into human hepatocellular carcinoma cell line BEL-7402 by liposome-encapsulated antisense IGF-Ⅰ oligodeoxynucleotides. MTT assay was used to detect cell proliferation The levels of AFP and CEA in the supernatant of the cultured cells were detected by radioimmunoassay.The changes of AFP and PCNA expression in the transfected cells were detected by immunohistochemistry and the changes of apoptosis were detected by Tunel method.Results: The A value of human hepatocellular carcinoma cell line BEL-7402 transfected with antisense IGF-Ⅰ oligonucleotide decreased from 0.44 ± 0.09 to 0.30 ± 0.07 (P <0.01), and the levels of AFP and CEA in supernatant decreased from 12.5 ± 2.2μg / L and 6.8 ± 2.3μg / L decreased to 2.5 ± 0.3μg / L and 2.2 ± 1.5μg / L (P <0.01, P <0.05) and the number of apoptotic cells increased from 16.4 ± 2.3% to 23.1 ± 3.7% P <0.05) .Conclusion: Transfection of antisense IGF-Ⅰ oligonucleotide into human hepatocellular carcinoma cell line BEL-7402 can decrease cell proliferation, decrease cell dedifferentiation and induce apoptosis of hepatoma cells.